Method of determining a volume of a liquid in a sample

Abstract

Disclosed herein are systems and methods for determining a volume of a liquid in a sample, for example, to calibrate a volume of a liquid in a sample to accurately determine the volume of an aliquot of the sample. Kits and software for implementing the systems and methods are also disclosed.

Description

Docket No. ABBTT-43595.601 SYSTEMS AND METHODS FOR DETERMINING BIOMARKERS IN BLOOD SAMPLES RELATED APPLICATION INFORMATION This application claims priority to U.S. Application No.63 / 674,136, filed on July 22, 2024, and U.S. Application No.63 / 680,844, filed on August 8, 2024, the contents of each of which are herein incorporated by reference. TECHNICALFIELDThe present disclosure relates to systems and methods for accurately and precisely determining small volumes of liquid of clinical samples. Kits and software for implementing the systems and methods are also disclosed. BACKGROUNDClinical diagnostic tests typically require an accurate and precise amount of sample because many biomarkers are recorded in terms of a measure of quantity per volume of sample (e.g., glucose mg / dL of plasma or RBC / uL of whole blood). Transfer of the sample to a test reaction container is usually conducted with highly accurate liquid transfer devices, such as volumetric pipettes. However, at very low volumes of sample (<10 uL), the manufacturing accuracy and precision of the transfer device can introduce volume variability that is unacceptable to laboratorians and regulators. This problem has plagued the development of very low clinical sample volume test development for decades. Thus, there is a need in the art for more accurate and precise methods of determining the delivered variable low volume of a sample. SUMMARY In one embodiment, the present disclosure relates to a method. In some embodiments, the method involves determining a volume of a liquid in a sample. The method comprises the steps of: (a) dispersing a predetermined number of microparticles per unit of a predetermined first volume of liquid into a first sample; (b) providing a second sample comprising an indeterminate second volume of liquid obtained from the predetermined first volume of liquid; and (c) determining the indeterminate second volume of liquid in the second sample based on the number of microparticles in the second sample. In some embodiments, the above method involves counting the number of microparticles in the first and / or second sample. In still further embodiments of the above method, the indeterminate second volume of liquid is determined by an equation, wherein the equation is:Docket No. ABBTT-43595.601 Volume = (number of microparticles counted in the second sample / predetermined number of microparticles dispersed in the first sample) x (predetermined first volume of the liquid in the first sample). In still further embodiments of the above method, the microparticles comprise inert microparticles. In some embodiments, the microparticles comprise monodisperse microparticles. In yet further embodiments, the microparticles comprise beads. In still further embodiments, the microparticles comprise colored or fluorescent beads. In yet further embodiments, the microparticles are counted using a flow cytometer, a fluorescence-based counter, an optical absorbed counter, a scattering counter, refractive index counter, a resistance-based counter, a capacitance-based counter, an inductance-based counter, an impedance-based counters, a nanopore sensor, a photoacoustic-based counter, a magnetic based counter, microscopy, particle image velocimetry, a thermal-based counter, or a vision-based automated counter. In still yet further embodiments, the microparticles comprise a uniform diameter of less than about 0.10 µm, less than about 0.20 µm, less than about 0.30 µm, less than about 0.40 µm, less than about 0.50 µm, less than about 0.60 µm, less than about 0.70 µm, less than about 0.80 µm, less than about 0.9 µm, less than about 1 µm, less than about 2 µm, less than about 3 µm, less than about 4 µm, or less than about 5 µm. In still further embodiments, the microparticles comprise a radionuclide. In some embodiments, the radionuclides are counted using a Geiger counter. In some embodiments, the radionuclide is selected from the group consisting of Co-57, Co-60, Cs-137, Eu-152, Ga-67, I-125, I- 132, In-111, Mo-99, P-32, Sm-153, Tc-99m, Tl-201. In some embodiments, the predetermined number of microparticles are provided in liquid. The liquid containing the predetermined number of microparticles is not critical. Any liquid that does not disrupt the sample can be used. For example, in some aspects, the liquid is a buffer or water. In some aspects, the predetermined number of microparticles are in the form of a liquid suspension. In some aspects, the predetermined number of microparticles are in the form of a homogeneous liquid suspension. In some further aspects, the predetermined number of microparticles is in a liquid suspension and is in Brownian motion. In still further aspects, the predetermined number of microparticles is in a homogeneous liquid suspension in Brownian motion.In some embodiments, the predetermined number of microparticles per unit of the volume of liquid is between 1 and 10000, 100 and 5000, 500 and 2000, 750 and 1500, 800 and 1400, 900 and 1100, or between 950 and 1050 microparticles per unit of the volume of liquid. In other embodiments, the predetermined number of microparticles per unit of the volume of liquid is about 1000 microparticles per µL of liquid. In yet other embodiments, the first sample comprises a bronchial lavage sample, a cerebrospinal fluid sample, a crevicular sample, an interstitial fluid sample, a saliva sample, a wholeDocket No. ABBTT-43595.601 blood sample, or a urine sample. In still yet other embodiments, the first sample comprises a suspension. In some embodiments, the predetermined first volume of liquid is between about 1µL and about 10 mL. In yet other embodiments, the indeterminate second volume of liquid is between about 10 nL and about 100 µL. In still further embodiments, the second sample is provided by aliquoting the indeterminate second volume of liquid from the first sample into a reaction vessel. In another embodiment, the method relates to calibrating a volume of a liquid in a sample to determine the volume of an aliquot of the sample. The method comprises the steps of: (a) dispersing a predetermined number of microparticles per unit volume of a predetermined first volume of liquid in a first sample, thereby calibrating the first volume of liquid in the first sample based on the number of microparticles per µL of liquid in the first sample; (b) aliquoting an indeterminate second volume of liquid from the first sample into a container to provide a second sample comprising a number of microparticles per µL of liquid that is proportionate to the number of microparticles per µL of liquid in the first sample; and (c) determining the volume of liquid in the second sample based on the number of microparticles in the second sample, wherein the volume of liquid in the second sample is equal to the volume obtained by multiplying the predetermined first volume of liquid in the first sample by the quotient of the number of microparticles in the second sample divided by the predetermined number of microparticles dispersed in the first sample. In another embodiment, the disclosure relates to a kit for calibrating a volume of a liquid in a sample to determine the volume of an aliquot of the sample. The kit can comprise: (a) a predetermined number of microparticles; and (b) a counting device to count an indeterminate number of microparticles in an aliquot of the sample; and (c) instructions for calibrating a predetermined volume of a liquid in a sample to determine the volume of an aliquot of the sample, wherein the instructions instruct a user of the kit to: (i) disperse the predetermined number of microparticles into a predetermined volume of liquid in a sample to calibrate the volume of liquid in the sample based on the number of microparticles per µL of liquid in the sample; and (ii) calculate the volume of liquid in an aliquot of the sample by multiplying the predetermined volume of liquid in the sample by the quotient of a counted number of the indeterminate number of microparticles in the aliquot of the sample divided by the predetermined number of microparticles dispersed in the sample. In some embodiments, the kit further comprises a sample collection device. In still other embodiments, the kit further comprises instructions for collecting a sample using the collectionDocket No. ABBTT-43595.601 device. In still further embodiments, the kit further comprises instructions for counting the indeterminate number of microparticles in the aliquot using the device. BRIEF DESCRIPTION OF THE DRAWINGS

[0001] The patent or application file contains drawings executed in color. Copies of this patent or patent application publication with color drawings will be provided by the Office upon request and payment of the necessary fee.

[0002] Having thus described the presently disclosed subject matter in general terms, reference will now be made to the accompanying Figures, which are not necessarily drawn to scale, and wherein:

[0003] FIG. 1 provides an example of how to calculate an unknown aliquot volume using a predetermined number of labeled microparticles or beads as described herein. Specifically, a homogeneous liquid suspension (e.g., water or buffer) containing a known number of labeled microparticles or beads is prepared (“Volume Calibrator Beads”). The volume of liquid containing the Volume Calibrator Beads is known. A known quantity (e.g., volume) of a first sample, such as a biological sample, is provided. As shown in FIG.1, the known volume of the first sample is 1000 µL. To the first sample, a small volume (the amount is not critical – it can be insignificant from a mathematical perspective) of the liquid containing the Volume Calibrator Beads is added to the first sample. The number of Volume Calibrator Beads added to the first sample from the liquid containing the Volume Calibrator Beads is known. As shown in FIG.1, the number of Volume Calibrator Beads in the small volume of the liquid containing the Volume Calibrator Beads is 10. The step of adding a small volume of liquid containing a known number of Volume Calibrator Beads to the first sample calibrates the first volume of liquid in the first sample based on the number of microparticles per µL of liquid in this sample (“Calibrated First Sample”). An unknown or indeterminate aliquot volume of the Calibrated First Sample is taken and used to create a second sample. As shown in Example 1, if 5 beads are counted in the second sample, the aliquot volume is determined by: 5 beads in the second sample / 10 beads in the Calibrated First Sample x 1000 µL (the volume of the first sample) to provide an aliquot volume of 500 µL. As shown in Example 2, if 3 beads are counted in the second sample, the aliquot volume is determined by: 3 beads in the second sample / 10 beads in the Calibrated First Sample x 1000 µL (the volume of the first sample) to provide an aliquot volume of 300 µL. As shown in Example 3, if 1 bead is counted in the second sample, the aliquot volume is determined by: 1 bead in the second sample / 10 beads in the Calibrated First Sample” x 1000 µL (the volume of the first sample) to provide an aliquot volume of 100 µL.Docket No. ABBTT-43595.601 DETAILED DESCRIPTION In an aspect, the present disclosure provides systems, methods and kits for determining a volume of a liquid in a sample, and more particularly to calibrating a volume of a liquid in a sample to precisely and accurately determine the volume of an aliquot of the sample. Small volumes of clinical samples (< 10 uL) are often difficult to aliquot accurately and precisely due to the imprecise dimensions of aliquoting devices such as manual or automated pipettes, microfluidic flow chambers, or other volumetric tools for the distribution of liquids. The work described herein illustrates that instead of controlling the flow of the sample, the inherent error in liquid delivery can be compensated by determining the volume of the liquid delivered independently using a volume calibrator. The use of volume calibrators can provide accurate low volumes of clinical samples to testing containers using inaccurate sample transfer devices yet permit high quality diagnostic tests to be performed providing less expensive tests than standard lab tests. The volume calibrators can also be used to calibrate an optical system that is used for blood cell counting. The systems, methods and kits of the first aspect of the disclosure involve dispersing a predetermined number of microparticles per unit of a predetermined first volume of liquid into a first sample; (b) providing a second sample comprising an indeterminate second volume of liquid obtained from the predetermined first volume of liquid; and (c) determining the indeterminate second volume of liquid in the second sample based on the number of microparticles in the second sample. Section headings as used in this section and the entire disclosure herein are merely for organizational purposes and are not intended to be limiting. 1. Definitions Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, will control. Preferred methods and materials are described below, although methods and materials similar or equivalent to those described herein can be used in practice or testing of the present disclosure. All publications, patent applications, patents and other references mentioned herein are incorporated by reference in their entirety. The materials, methods, and examples disclosed herein are illustrative only and not intended to be limiting. The terms “comprise(s),” “include(s),” “having,” “has,” “can,” “contain(s),” and variants thereof, as used herein, are intended to be open-ended transitional phrases, terms, or words that do not preclude the possibility of additional acts or structures. The singular forms “a,” “an” and “the” include plural references unless the context clearly dictates otherwise. The present disclosure also contemplates other embodiments “comprising,” “consisting of” and “consisting essentially of,” the embodiments or elements presented herein, whether explicitly set forth or not.Docket No. ABBTT-43595.601 For the recitation of numeric ranges herein, each intervening number there between with the same degree of precision is explicitly contemplated. For example, for the range of 6-9, the numbers 7 and 8 are contemplated in addition to 6 and 9, and for the range 6.0-7.0, the number 6.0, 6.1, 6.2, 6.3, 6.4, 6.5, 6.6, 6.7, 6.8, 6.9, and 7.0 are explicitly contemplated. An “amount” as used herein refers to a quantity specified (e.g., high or low) or a number e.g., where the number is a level, such as a position on a real or imaginary scale of amount or quantity, or a concentration, such as, for example, a relative amount of a given substance contained within a solution or in a particular volume of space, e.g., the amount of solute per unit volume of solution. “Amplifying” or “amplification” in the context of nucleic acids refers to the production of multiple copies of a polynucleotide, or a portion of the polynucleotide, typically starting from a small amount of the polynucleotide (e.g., a single polynucleotide molecule), where the amplification products or amplicons are generally detectable. Amplification of polynucleotides encompasses a variety of chemical and enzymatic processes. Generation of multiple DNA copies from one or a few copies of a target or template DNA molecule during a polymerase chain reaction (PCR), isothermal reaction, or a ligase chain reaction (LCR) are forms of amplification. Amplification is not limited to the strict duplication of the starting molecule. For example, the generation of multiple cDNA molecules from a limited amount of RNA in a sample using reverse transcription (RT)-PCR is a form of amplification. Furthermore, the generation of multiple RNA molecules from a single DNA molecule during the process of transcription is also a form of amplification. An “analog assay” as used herein refers to an assay in which the presence of and / or amount of an analyte in a test sample is determined by measuring the total signal produced (e.g., fluorescence, color, etc.) by the analyte in an entire reaction mixture (e.g., in a reaction vessel). In an analog assay, the noise is indistinguishable from the signal. An example of an analog assay is an assay in which the presence of and / or amount of an analyte is determined by measuring the total signal produced from a plurality of beads or microparticles contained in a reaction vessel. “Analyte”, “analyte of interest”, “biomarker” or “biomarker of interest” as used interchangeably herein, refers to a naturally occurring or synthetic biological molecule found in an organism, in which the varying concentrations provide information useful in predicting the risk, occurrence, or severity of a disease or condition. In some embodiments, the analyte is a biomolecule. Non-limiting examples of biomolecules include macromolecules such as proteins (e.g., peptides, polypeptides, protein fragments, protein complexes, fusion proteins, recombinant proteins, phosphoproteins, glycoproteins, lipoproteins), lipids (e.g., fatty acids, cholesterol, triglycerides, phospholipids), or carbohydrates (e.g., glucose, glycogen, glycoproteins, glycolipids). In certain instances, the analyte may be hormones (e.g., insulin, cortisol, estrogen, testosterone), antibodies, growth factors (e.g., epidermal growth factor, insulin-like growth factor), cytokines (e.g., interleukins,Docket No. ABBTT-43595.601 tumor necrosis factor, interferons), enzymes (e.g., alanine transaminase, creatine kinase), receptors (e.g., neural receptors, hormonal receptors, nutrient receptors, cell surface receptors) and their ligands, cancer markers (e.g., prostate-specific antigen (PSA), tumor necrosis factor-alpha (TNF-alpha)), markers of myocardial infarction (e.g., troponin, creatine kinase), toxins (e.g., heavy metals, environmental pollutants), drugs (e.g., drugs of addiction, pharmaceuticals), metabolic agents (e.g., vitamins, minerals), non-naturally occurring polymorphisms (e.g., genetic variations, mutations) and the like. In some embodiments, the analyte may be a post-translationally modified protein (e.g., phosphorylated, methylated, glycosylated protein) and the first or the second binding member may be an antibody specific to a post-translational modification. A modified protein may be bound to a first binding member immobilized on a solid support where the first binding member binds to the modified protein but not the unmodified protein. In other embodiments, the first binding member may bind to both the unmodified and the modified protein, and the second binding member may be specific to the post-translationally modified protein. In yet other embodiments, the analyte may be a cell, such as, a circulating tumor cell, stem cells, white blood cells (leukocytes), red blood cells (erythrocytes), platelets (thrombocytes), immune cells (e.g., lymphocytes, macrophages, neutrophils), endothelial cells, epithelial cells (e.g., from the skin, urinary tract, gastrointestinal tract), mesenchymal cells, neuronal cells, pathogenic bacteria, viruses (including retroviruses, herpesviruses, adenoviruses, lentiviruses, Filoviruses (ebola), hepatitis viruses (e.g., A, B, C, D, and E, including surface antigens, core antigens, etc.), human immunodeficiency viruses (e.g., HIV-1, HIV-2), paroviruses, Esptein-Barr Virus, HPV), parasites (e.g., babesia, Trypanosoma cruzi), spores, etc. A non-limiting list of analytes include: Anti-Mullerian Hormone (AMH), Autoantibody to CD25, CXCL13, Dkk-3 (Semen), IL-12p40, IL-8, p14 Endocan Fragment, SARS-CoV-2 IgA Antibody, SARS-CoV-2 IgG Antibody, SARS-CoV-2 IgM Antibody, Secretory Gelsolin (pGSN), Secretogranin II, ACE2, Albumin, Albuminuria, Alpha-Amylase, Apo H, B-2 Microglobulin Levels, CA 24-2, Carcinoembryonic Antigen (CEA), Ceruloplasmin, Chloride, Cholinesterase, Copper, CRP (C-Reactive Protein), Fibrinogen Alpha Chain (FGA), Homocysteine, IL-18 (Interleukin 18), IL-6 (Interleukin 6), LD (Lactate Dehydrogenase), L-FABP (Liver Fatty Acid Binding Protein), Lipase, Microalbuminuria, Neutrophil Gelatinase-Associated Lipocalin (NGAL), Osteopontin, PGC-1a (Peroxisome Proliferator-Activated Receptor Gamma Coactivator-1 Alpha), PKR (Proapoptotic Kinase R) and its phosphorylated PKR (pPKR), PCT (Procalcitonin), Pepsinogen I, Pepsinogen II, Pro-SFTPB, PTH (Parathyroid Hormone), sIL-2, SHBG (Sex Hormone-Binding Globulin), Thioredoxin, TNFα (Tumor Necrosis Factor-Alpha), TSH (Thyroid Stimulating Hormone), Vitamin D-Binding Protein, Alpha-synuclein, BARF1 (BamH1-A Reading Frame 1), Kidney Injury Molecule- 1 (KIM-1), Laminin gamma (e.g., laminin gamma subunit 1, laminin gamma subunit 2, lamininDocket No. ABBTT-43595.601 gamma subunit 3), LMP1 (Latent Membrane Protein 1), Neurofilament light chain, Tau protein, UCH-L1 (Ubiquitin C-Terminal Hydrolase-L1), Alkaline Phosphatase, Amylase, AST (Aspartate Aminotransferase), Calcium, Cholesterol, CK (Creatine Kinase), CO2 (Carbon Dioxide), Creatinine, Direct LDL (Direct Low-Density Lipoprotein), GGT (Gamma-Glutamyl Transferase), HDL (High- Density Lipoprotein), Iron, LDL (Low-Density Lipoprotein), Magnesium, Potassium (K), Sodium (Na), Triglycerides, Uric Acid, Akt (Protein Kinase B), Amphiregulin, ANXA7 (Annexin A7), AR (Androgen Receptor), BRAF (v-Raf Murine Sarcoma Viral Oncogene Homolog B), CDKN1B (Cyclin-Dependent Kinase Inhibitor 1B), cMYC (MYC Proto-Oncogene), CTNNB1 (Catenin Beta-1), EGFR (Epidermal Growth Factor Receptor), EPHB2 (Ephrin Type-B Receptor 2), ESR1 (Estrogen Receptor 1), ESR2 (Estrogen Receptor 2), FOXO3A (Forkhead Box O3), FRAP1 (Mechanistic Target of Rapamycin Complex 1), FRS2 (Fibroblast Growth Factor Receptor Substrate 2), Gab2 (GRB2- Associated Binding Protein 2), GFAP (Glial Fibrillary Acidic Protein), Grb2 (Growth Factor Receptor-Bound Protein 2), GREB1 (Growth Regulation By Estrogen In Breast Cancer 1), HER2 (Human Epidermal Growth Factor Receptor 2), HER3 (Human Epidermal Growth Factor Receptor 3), HER4 (Human Epidermal Growth Factor Receptor 4), IGF-IR (Insulin-Like Growth Factor 1 Receptor), IL6R (Interleukin-6 Receptor), KLF6 (Kruppel Like Factor 6), KRAS (Kirsten Rat Sarcoma Viral Oncogene Homolog), LZTS1 (Leucine Zipper Tumor Suppressor 1), MAP2K1 (Mitogen-Activated Protein Kinase Kinase 1), MEK (Mitogen-Activated Protein Kinase Kinase), MIG-6 (Mitogen-Inducible Gene 6 Protein), MKI67 (Marker of Proliferation Ki-67), mTOR (Mechanistic Target of Rapamycin), MUC4 (Mucin 4, Cell Surface Associated), NEDD4-1 (Neural Precursor Cell Expressed Developmentally Down-Regulated Protein 4-1), NKX3-1 (NK3 Homeobox 1), NRG1 (Neuregulin 1), Parkin, PDK-1 (3-Phosphoinositide-Dependent Protein Kinase-1), PGR (Progesterone Receptor), PHLPP (PH Domain and Leucine-Rich Repeat Protein Phosphatase), PIK3CA (Phosphatidylinositol-4,5-Bisphosphate 3-Kinase Catalytic Subunit Alpha), PITX2 (Paired- Like Homeodomain Transcription Factor 2), PPP1R1B (Protein Phosphatase 1 Regulatory Inhibitor Subunit 1B), PRDX6 (Peroxiredoxin-6), PTEN, PTEN 1 (Phosphatase and Tensin Homolog 1), PXN (Paxillin), S6K (Ribosomal Protein S6 Kinase), SHIP (Src Homology 2 Domain-Containing Inositol Phosphatase), Src (Proto-Oncogene Tyrosine-Protein Kinase Src), TOPO II (DNA Topoisomerase II), TSC1 (Tuberous Sclerosis 1), TSC2 (Tuberous Sclerosis 2), Tumor necrosis factor-alpha receptors, VEGF (Vascular Endothelial Growth Factor), Aβ42 (Amyloid Beta-Protein 42), CK-MB (Creatine Kinase-MB), Anti-CCP (Anti-Cyclic Citrullinated Peptide), Anti-Tg (Anti-Thyroglobulin Antibody), Anti-TPO (Anti-Thyroid Peroxidase Antibody), ASO (Antistreptolysin O), C3 (Complement Component 3), C4 (Complement Component 4), D-Dimer, HIV Ag / Ab Combo (Human Immunodeficiency Virus Antigen / Antibody Combination), RF (Rheumatoid Factor), DJ-1, Leucine- rich repeat kinase 2, Mutated ATP13A2, Prion protein, PTEN induced putative kinase 1, Troponin (such as troponin I, troponin T , macrotroponin and / or troponin C), AFP (Alpha-Fetoprotein), CA 125 (MUC16), CA 15-3, CA 19-9, CCNI (Cyclin I), CMV (Cytomegalovirus), CYFRA21-1, EGRFDocket No. ABBTT-43595.601 (Epidermal Growth Factor Receptor), FGF19 (Fibroblast Growth Factor 19), HE-4 (Human Epididymis Protein 4), NSE (Neuron-Specific Enolase), p-ANCA (Perinuclear Anti-Neutrophil Cytoplasmic Antibody), PIVKA, PIVKA-II (Protein Induced By Vitamin K Absence-II), Pro-SFTPB, PSA (Prostate Specific Antigen), Rubella, SCC (Squamous Cell Carcinoma Antigen), Toxo IgG, Toxo IgM, Beta-hcG (Beta Human Chorionic Gonadotropin), Botulinum toxins, Clostridium difficile toxins A and B, DHEA-S (Dehydroepiandrosterone Sulfate), Diphtheria toxin, E. coli enterotoxins (heat-labile exotoxin, heat-stable enterotoxin), Estradiol, Fetuin-A, FSH (Follicle-Stimulating Hormone), Glycosylated hemoglobin (HbA1c), Hemoglobin A1c, IL1α (Interleukin 1 alpha), Influenza HA antigen, Insulin, LH (Luteinizing Hormone), Myeloperoxidase (MPO), NF-1 (Neurofibromin 1), Plasma C-peptide, PlGF (Placental Growth Factor), Pro-GFP, Prolactin, S100β, sFlt-1 (Soluble Fms-Like Tyrosine Kinase-1), Testosterone, Tetanus toxin, Thymosin 1315, ALT (Alanine Aminotransferase), Bile Acids, Total, Bilirubin, Bilirubin, Direct, Bilirubin, Total, Calprotectin, DUTPase (Deoxyuridine Triphosphatase), Glucose, Lactic Acid, Lactoferrin, Protein, Shiga toxin, Shiga-like toxin I, Shiga-like toxin II, Total Protein, Urea Nitrogen, VDAC1 (Voltage- Dependent Anion Channel 1), Wilm's Tumor-1 protein, Amphetamine / Methamphetamine, Barbiturates, Benzodiazepines, Benzodiazepines (Serum), Cannabinoids, Cocaine, Ecstasy, Ethanol, Methadone, Opiates, PCP (Phencyclidine), propoxyphene, salicylate, or tricyclic antidepressants. In another aspect, the analyte is cardiac myosin binding protein C, myoglobin, brain natriuretic peptide and derivatives thereof (e.g., preproBNP, proBNP, NT-proBNP, BNP). In still other aspects, the analyte is digoxin, gentamicin, methotrexate, phenobarbital, phenytoin, theophylline, valproic acid, vancomycin, B12, C-peptide, cortisol, ferritin, folate, insulin, vitamin D folate RBC, Free T3, Free T4, Thyroglobulin, Total T3, Total T4, TRAb, cyclosporine, sirolimus, tacrolimus, DDP4, or periostin. Examples of nucleic acid aptamers include: drugs of abuse (e.g. cocaine), protein analytes (including, but not limited to, Nucleolin, nuclear factor-kB essential modulator (NEMO), CD-30, protein tyrosine kinase 7 (PTK7), vascular endothelial growth factor (VEGF), MUC1 glycoform, immunoglobulin µ Heavy Chains (IGHM), Immunoglobulin E, αvβ3 integrin, α-thrombin, NF-κB, E2F transcription factor, HER3, Plasminogen activator inhibitor, Tenascin C, CXCL12 / SDF-1, prostate specific membrane antigen (PSMA), gastric cancer cells, HGC-27); cells (including, but not limited to, non-small cell lung cancer (NSCLC), colorectal cancer cells, (DLD-1), H23 lung adenocarcinoma cells, Ramos cells, T-cell acute lymphoblastic leukemia (T-ALL) cells, CCRF-CEM, acute myeloid leukemia (AML) cells (HL60), small-cell lung cancer (SCLC) cells, NCIH69, human glioblastoma cells, U118-MG, PC-3 cells, HER-2-overexpressing human breast cancer cells, SK-BR- 3, pancreatic cancer cell line (Mia-PaCa-2)); and infectious agents (including, but not limited to, Mycobacterium tuberculosis, Staphylococcus aureus, Shigella dysenteriae, Escherichia coli O157:H7, Campylobacter jejuni, Listeria monocytogenes, Pseudomonas aeruginosa, Salmonella 08, Salmonella enteritidis).Docket No. ABBTT-43595.601 Examples of protein or peptide aptamers include: HBV core capsid protein, CDK2, E2F transcription factor, Thymidylate synthase, Ras, EB1, and Receptor for Advanced Glycated End products (RAGE). Aptamers, and use and methods of production thereof are reviewed in e.g., Shum et al., J Cancer Ther.20134:872; Zhang et al., Curr Med Chem.2011; 18:4185; Zhu et al., Chem Commun (Camb). 201248:10472; Crawford et al., Brief Funct Genomic Proteomic.20032:72; Reverdatto et al., PLoS One 20138:e65180. “Bead,” “microparticle,” and “particle” are used herein interchangeably and refer to a substantially spherical solid support. Microparticles that can be used herein can be any type known in the art. For example, the bead or microparticle can be a magnetic bead or magnetic particle. Magnetic beads / particles may be ferromagnetic, ferrimagnetic, paramagnetic, superparamagnetic or ferrofluidic. Exemplary ferromagnetic materials include Fe, Co, Ni, Gd, Dy, CrO2, MnAs, MnBi, EuO, and NiO / Fe. Examples of ferrimagnetic materials include NiFe2O4, CoFe2O4, Fe3O4 (or FeO.Fe2O3). Beads can have a solid core portion that is magnetic and is surrounded by one or more non-magnetic layers. The microparticle comprises a glass microsphere or bead. The disclosure contemplates the use of both solid and hollow glass microspheres or beads. In some embodiments, the microparticle comprises a polymer microsphere. Exemplary polymer microspheres can include, without limitation, polyethylene microspheres, poly(methyl methyl acrylate) (PMMA) microspheres and polypropylene microspheres. In other embodiments, the microparticle comprises a ceramic microsphere or nanosphere. In an exemplary embodiment, the microparticle comprises a silica microsphere or nanosphere. In an embodiment, the microparticle comprises a yttria stabilized zirconia microsphere. In an embodiment, the microparticle comprises a cerium stabilized zirconia microsphere. In an embodiment, the microparticle comprises a zirconium silicate bead. In some embodiments, the microparticle comprises a metal microsphere. In an embodiment, the microparticle comprises a stainless-steel microsphere. In an embodiment, the microparticle comprises a titanium spherical microparticle. In other embodiments, the microparticle comprises a fluorescent microsphere that emits bright colors when illuminated by UV light providing contrast and visibility of microspheres relative to background materials. The fluorescent microspheres are particularly useful in methods in which a flow cytometer is used to count the microparticles. Fluorescent microspheres can also be detected using acoustic and optical analytical instruments for counting microparticles. In one embodiment, the microparticle comprises a fluorescent polymer microsphere. The skilled artisan will appreciate that fluorescent polymer microspheres are readily available in various particle sizes, colors, densities, and excitation and emission wavelengths. They can also be stored at ambient temperatures or dispersed in aqueous media without diminishing their fluorescence. In one embodiment, the fluorescent microparticle comprises a fluorescent powder. In another embodiment, the fluorescent microparticle comprises a glass, polymer or ceramic microsphere coated with aDocket No. ABBTT-43595.601 fluorescent coating that can be colored, fluorescent in light, and display bright fluorescence when exposed to UV light. The skilled artisan will appreciate that certain microparticles (e.g., polyethylene microspheres) that are hydrophobic may require a surfactant for proper suspension in a volume of liquid (e.g., sample or aliquot of a sample). In some embodiments, the microparticles used in the methods are hydrophilic. In other embodiments, the method further comprises dispersing a surfactant in the volume of liquid with the predetermined number of microparticles. The surfactant can be combined with the microparticles, for example, in a buffer, prior to dispersing the predetermined number of microparticles in the volume of liquid. The surfactant can also be added simultaneously or sequentially with the microparticles to the predetermined volume of liquid. The skilled artisan will also appreciate that the microparticles or beads can have a variety of suitable densities. The microparticle (e.g., bead) density can be selected to ensure that the microparticles remain in suspension in the sample. In some embodiments, the microparticles have a density less than that of water. The microparticles can be colored microspheres or beads. The colored microparticles can come in various colors, such as white, green, red, black, grey, orange, yellow, blue, pink, or violate. Other colors could also be used. The microparticles can also be coated microparticles. For example, the microparticles can be coated with aluminum, a fluorescent coating, a fluorochemical coating, a gold coating, a nickel plating, a silane coating, a silver coating, or a titanium oxide coating. In some embodiments, the microparticle comprises a radionuclide. The skilled person will appreciate that radionuclide are effective in applications where the counting instrument is a Geiger counter. Radionuclides can also be useful in analytical methods involving gas proportional detectors for alpha and beta-particle counting (GP), sodium iodide or high-resolution germanium detectors for gamma detection and spectrometry, low-energy gamma- or X-ray detectors, kinetic phosphorimeter analysis, mass spectrometric analyses (e.g., inductively coupled plasma-mass spectrometry, thermal ionization mass spectrometry, accelerator mass spectrometry), and neutron activation. The skilled artisan will appreciate that the radionuclide used and instrument for its detection depend on the type of emitter of the radionuclide (e.g., whether it’s an alpha-, beta-, or gamma-emitter). The radionuclide can be used to calibrate a volume of liquid (e.g., an aliquot) based on the count of the radionuclide in the sample taken (e.g., aliquot). Because the ranges of measurement and calibration (for example, from μL to pL in volume or from μL / min to pL / min in flow rate) are different, various nuclide species (for example, the ion state or a complex form) and concentrations (for example, different dilution folds) of different liquid state (for example, hydrophilic or hydrophobic) calibration sources must be selected as usable standards for measurement and calibration, and the liquid state standards for measurement may be adjusted to a whole bloodDocket No. ABBTT-43595.601 suspension system or water solution system. Standards are selected according to factors such as half- life period of nuclides, specific activity (TBq / g), atomic mass (amu) and mode of decay. In addition, availability of standards shall also be considered. The data of some alternative nuclides is listed in Table 1. Table 1. Data of Exemplary RadionuclidesExemplary radionuclides of use herein include, without limitation, Co-57, Co-60, Cs-137, Eu-152, Ga-67, I-125, I-132, In-111, Mo-99, P-32, Sm-153, Tc-99m, Tl-201 or combinations thereof. The size of the microparticles used as volume calibrators in the systems, methods and kits described herein can vary. Preferably, the microparticles have a substantially uniform diameter of less than about 0.10 µm, less than about 0.20 µm, less than about 0.30 µm, less than about 0.40 µm, less than about 0.50 µm, less than about 0.60 µm, less than about 0.70 µm, less than about 0.80 µm, less than about 0.9 µm, less than about 1 µm, less than about 2 µm, less than about 3 µm, less than about 4 µm, or less than about 5 µm. The skilled artisan will appreciate, however, that the microparticles should be sized to support diffusion of the microparticles in solution. In some instances, for example, for larger sized microparticles, the disclosure contemplates agitation of the sample (e.g., aliquot) to ensure that the microparticles are sufficiently dispersed throughout the sample. “Capillary blood sample” as used herein refers to a sample of blood from the capillaries which is obtained (e.g., extracted) through the skin (and not the veins) of a subject using a syringe, needle, or any other suitable device or combination thereof. For example, a whole blood sample can be obtained from the skin on the fingers and / or toes, a hand, a foot (including the heel), an earlobe, a location on the arms and / or legs, chest, back, head, or any combinations thereof. In someDocket No. ABBTT-43595.601 embodiments, the capillary blood sample is whole blood, serum or plasma. In yet other embodiments, the capillary blood sample contains predominantly capillary blood, but may also contain or comprise a small amount or percentage of interstitial fluid. “Cartridge” as used herein refers to a hollow container and / or chip that comprises one or more substances and / or components (e.g., a liquid, reagents (e.g., antibodies and / or antigens), and / or a particle (e.g., a bead, or microparticle)) for insertion into an apparatus (e.g., a point-of-care device). In some aspects, a cartridge has one or more apertures. In some aspects, a cartridge is a microfluidic cartridge. “Communicating the amount” as used herein, refers to communicating, as described herein, the amount, in terms of presence (i.e., a qualitative measure), or in terms of a level or levels (i.e., a quantitative measure), such that “amount” refers to presence or level. In some aspects, “communicating the amount” refers to communicating the presence (e.g., qualitative measure) of an analyte or biomarker. In other aspects, “communicating the amount” refers to communicating the level (e.g., quantitative measure) of an analyte or biomarker. "Colorimetric" refers to the physical description and quantification of chromatography, including human color perception spectroscopy (e.g., visible spectroscopy). In some embodiments, colorimetric assays are particularly useful when quantification is not required and there are no expensive detection devices. In certain embodiments, detection of a color change may be performed by visual inspection by a user (e.g., the person performing the assay). Because the colorimetric assay can be detected by visual inspection, the user can check for a detectable change in the color of the reaction, or the assay can be run in parallel with one or more controls (positive or negative) that replicate the color of the comparable reaction. In some embodiments, calibrated colorimetric measurements may be used to quantify the amount of the target. Typically, colorimetric analysis involves determining the presence / absence, level or concentration of an analyte (e.g., a chemical element or compound) in a sample (e.g., a solution) with the aid of a color-developer. It is suitable for organic and inorganic compounds and can be used with or without an enzymatic reaction step. Typically, the desired device is a colorimeter, one or more cuvettes and a suitable color developer. The process may be automated, for example by using an automated analyzer or by flow injection analysis. In particular embodiments, the colorimeter may be adapted for use with a plate reader to accelerate analysis and reduce waste streams. “Component,” “components,” or “at least one component,” refer generally to a specific binding member (e.g., a capture agent, e.g., capture antibody or aptamer), a detection or conjugate a calibrator, a control, a sensitivity panel, a container, a buffer, a diluent, a salt, an enzyme, a co-factor for an enzyme, a detection agent, a pretreatment reagent / solution, a substrate (e.g., as a solution), a stop solution, and the like that can be included in a kit for assay of a test sample, such as a patient whole blood, serum or plasma sample, in accordance with the methods described herein and otherDocket No. ABBTT-43595.601 methods known in the art. Some components can be in solution or lyophilized for reconstitution for use in an assay. "Decentralize”, “Decentralized”, or “Decentralization”, as used interchangeably herein, refers to, in the context of testing, the performance of one or more medical tests and / or assays outside of a traditional medical setting (e.g., a hospital, physician office, stand alone lab site, etc.) to one or more places such as urgent care clinics, retail clinics, pharmacies, grocery stores or convenience stores, residences (e.g., homes, apartments, etc.), workplaces, and / or government offices (e.g., U.S. Transportation and Safety Authority), etc. “Hybrid-decentralization” or “hybrid-decentralized” refers to situations in which a subject or patient collects a sample at a residence and / or workplace and ships the sample to a laboratory, avoiding a professional collection site (such as a hospital, physician’s office, or stand-alone sample collection or lab site). “Determined by an assay” is used herein to refer to the determination of a reference level by any appropriate assay. The determination of a reference level may, in some embodiments, be achieved by an assay of the same type as the assay that is to be applied to the sample from the subject (for example, by an immunoassay, clinical chemistry assay, a single molecule detection assay, protein immunoprecipitation, immunoelectrophoresis, chemical analysis, SDS-PAGE and Western blot analysis, or protein immunostaining, electrophoresis analysis, a protein assay, a competitive binding assay, a functional protein assay, or chromatography or spectrometry methods, such as high- performance liquid chromatography (HPLC) or liquid chromatography–mass spectrometry (LC / MS)). The determination of a reference level may, in some embodiments, be achieved by an assay of the same type and under the same assay conditions as the assay that is to be applied to the sample from the subject. As noted herein, this disclosure provides exemplary reference levels (e.g., calculated by comparing reference levels at different time points). It is well within the ordinary skill of one in the art to adapt the disclosure herein for other assays to obtain assay-specific reference levels for those other assays based on the description provided by this disclosure. For example, a set of training samples comprising samples obtained from human subjects known to have a disease or condition and samples obtained from human subjects known not to have the disease or condition may be used to obtain assay-specific reference levels. It will be understood that a reference level “determined by an assay” and having a recited level of “sensitivity” and / or “specificity” is used herein to refer to a reference level which has been determined to provide a method of the recited sensitivity and / or specificity when said reference level is adopted in the methods of the disclosure. It is well within the ordinary skill of one in the art to determine the sensitivity and specificity associated with a given reference level in the methods of the disclosure, for example by repeated statistical analysis of assay data using a plurality of different possible reference levels. “Digital assay” as used herein refers to an assay in which an analyte is bound or captured and a molecule of the analyte segregated and interrogated (e.g., to detect the presence and / or amountDocket No. ABBTT-43595.601 of the analyte in a sample). In a digital assay, noise is separated from signal. In a digital assay, the results are assigned a value of 1 or 0. Examples of digital assays include one or more of the following (which may overlap but are not mutually exclusive): multiplexed nucleic acid amplification (e.g., digital PCR or isothermal amplification), single molecule detection assay, a nanowell assay, a multiplexed immunoassay (e.g., sandwich immunoassay), a single molecule enzyme linked immunosorbent assay, a direct capture counting assay, etc. As used herein, “digital microfluidics (DMF),” “digital microfluidic module (DMF module),” or “digital microfluidic device (DMF device)” as used interchangeably herein refer to a module, device, or system (e.g., micro-total analysis systems, or biomedical (or biological) microelectromechanical systems (BIOMEMs)) that utilizes digital or droplet-based microfluidic techniques to provide for manipulation of discrete and small volumes of liquids in the form of droplets. Digital microfluidics uses the principles of emulsion science to create fluid-fluid dispersion into channels (principally water-in-oil emulsion). It allows the production of monodisperse drops / bubbles or with a very low polydispersity. Digital microfluidics is based upon the micromanipulation of discontinuous fluid droplets within a reconfigurable network. Complex instructions can be programmed by combining the basic operations of droplet formation, translocation, splitting, and merging. Digital microfluidics operates on discrete volumes of fluids that can be manipulated by binary electrical signals. By using discrete unit-volume droplets, a microfluidic operation may be defined as a set of repeated basic operations, i.e., moving one unit of fluid over one unit of distance. Droplets may be formed using surface tension properties of the liquid. Actuation of a droplet is based on the presence of electrostatic forces generated by electrodes placed beneath the bottom surface on which the droplet is located. Different types of electrostatic forces can be used to control the shape and motion of the droplets. One technique that can be used to create the foregoing electrostatic forces is based on dielectrophoresis which relies on the difference of electrical permittivities between the droplet and surrounding medium and may utilize high-frequency AC electric fields. Another technique that can be used to create the foregoing electrostatic forces is based on electrowetting, which relies on the dependence of surface tension between a liquid droplet present on a surface and the surface on the electric field applied to the surface. Digital microfluidics can be performed a cartridge a chip (e.g., lab-on-chip, biochip), or a combination thereof. As used herein, the term “digital PCR” (dPCR) refers to a nucleic acid amplification method for the quantitation and analysis of target nucleic acid sequences by partitioning the reaction into a sufficient number of reaction subvolumes so that the target is in limiting dilution, i.e., producing a sufficient number of reaction subvolumes with zero target copies so as to allow the application of Poisson statistics.Docket No. ABBTT-43595.601 As used herein, “digital PCR methods” refer to the use of one or more of the steps typically employed by dPCR, but does not require that the “digital assumption” be achieved, namely, the assumption that any reaction subvolume that produced target amplicons had a single initial copy of the target sequence. As used herein, the term “droplet” refers to a small volume of liquid that is immiscible with its surroundings (e.g. gases, liquids, surfaces, etc.). A droplet may reside upon a surface, be encapsulated by a fluid with which it is immiscible (e.g. the continuous phase of an emulsion, a gas (e.g. air, nitrogen)), or a combination thereof. A droplet is typically spherical or substantially spherical in shape, but may be non-spherical. The shape of an otherwise spherical or substantially spherical droplet may be altered by deposition onto a surface or constriction in a capillary channel of smaller diameter. A droplet may be a “simple droplet” or a “compound droplet,” wherein one droplet encapsulates one or more additional smaller droplets. The volume of a droplet and / or the average volume of a set of droplets provided herein is typically less than about one microliter (e.g.0.1 μL ... 10 nL ...1 nL ...0.1 nL ...10 μL ...1 μL ...0.1 μL ...100 fL ...10 fL ...1 fL). The diameter of a droplet and / or the average diameter of a set of droplets provided herein is typically less than about one millimeter (e.g.1 mm ...100 μm ...10 μm ...1 μm). Droplets may be formed by any suitable technique (e.g. emulsification, microfluidics, injection etc.) and may be monodisperse (e.g., substantially monodisperse) or polydisperse. “Dynamic range” as used herein refers to range over which an assay readout is proportional to the amount of target molecule or analyte in the sample being analyzed. “Higher throughput assay analyzer” or a “non-point-of-care device”, as used interchangeably herein, refers to a device that is not a point-of-care device or a single use device. A higher throughput assay analyzer or non-point-of-care device refers to any device that does not meet any of the limitations of a point-of-care or a single use device as defined herein. In some embodiments, a “higher throughput assay analyzer” or “non-point-of-care device” refers to an instrument that: (a) may be a relatively large instrument compared to a hand-held point-of-care device, e.g., such as ranging in size from that of a tabletop instrument (e.g., typically considered low- or medium-throughput) to a large room-size or multiple-room-size instrument (e.g., typically considered high throughput); (b) is not a handheld instrument; (c) is capable of performing an assay on more than one clinical sample simultaneously; and (d) any combination of (a)-(c). A higher throughput assay analyzer may be a clinical chemistry analyzer, an immunoassay analyzer, or a combination thereof. Exemplary higher throughput assay analyzers or non-point-of-care devices include, for example, the ARCHITECT or Alinity platforms produced by Abbott Laboratories. As used herein the term “hydrophilic”, such as in reference to a “hydrophilic material” (e.g., membrane, microparticle, film, etc.) refers to those materials having a water contact angle of less than about 40 degrees.Docket No. ABBTT-43595.601 As used herein the term “hydrophobic”, such as in reference to a “hydrophobic material” (e.g., membrane, microparticle, film, etc.) refers to those materials having a water contact angle greater than about 80 degrees. “Label” and “detectable label” as used interchangeably herein refer to a moiety attached to an antibody or an analyte to render the reaction between the antibody and the analyte detectable, and the antibody or analyte so labeled is referred to as “detectably labeled.” A label can produce a signal that is detectable by visual or instrumental means. Various labels include signal-producing substances, such as chromagens, fluorescent compounds, chemiluminescent compounds, radioactive compounds, and the like. Representative examples of labels include moieties that produce light, e.g., acridinium compounds, and moieties that produce fluorescence, e.g., fluorescein. Other labels are described herein. In this regard, the moiety itself may not be detectable but may become detectable upon reaction with yet another moiety. Use of the term “detectably labeled” is intended to encompass such labeling. Any suitable detectable label as is known in the art can be used. For example, the detectable label can be a radioactive label (such as 3H, 14C, 32P, 33P, 35S, 90Y, 99Tc, 111In, 125I, 131I, 177Lu, 166Ho, and 153Sm), an enzymatic label (such as horseradish peroxidase, alkaline peroxidase, glucose 6-phosphate dehydrogenase, and the like), a chemiluminescent label (such as acridinium esters, thioesters, or sulfonamides; luminol, isoluminol, phenanthridinium esters, and the like), a fluorescent label (such as fluorescein (e.g., 5-fluorescein, 6-carboxyfluorescein, 3’6- carboxyfluorescein, 5(6)-carboxyfluorescein, 6-hexachloro-fluorescein, 6-tetrachlorofluorescein, fluorescein isothiocyanate, and the like)), rhodamine, phycobiliproteins, R-phycoerythrin, quantum dots (e.g., zinc sulfide-capped cadmium selenide), a thermometric label, or an immuno-polymerase chain reaction label. An introduction to labels, labeling procedures and detection of labels is found in Polak and Van Noorden, Introduction to Immunocytochemistry, 2nd ed., Springer Verlag, N.Y. (1997), and in Haugland, Handbook of Fluorescent Probes and Research Chemicals (1996), which is a combined handbook and catalogue published by Molecular Probes, Inc., Eugene, Oregon. A fluorescent label can be used in FPIA (see, e.g., U.S. Patent Nos.5,593,896, 5,573,904, 5,496,925, 5,359,093, and 5,352,803, which are hereby incorporated by reference in their entireties). An acridinium compound can be used as a detectable label in a homogeneous chemiluminescent assay (see, e.g., Adamczyk et al., Bioorg. Med. Chem. Lett.16: 1324-1328 (2006); Adamczyk et al., Bioorg. Med. Chem. Lett.4: 2313-2317 (2004); Adamczyk et al., Biorg. Med. Chem. Lett.14: 3917-3921 (2004); and Adamczyk et al., Org. Lett.5: 3779-3782 (2003)). Other labels that can be used include a tag attached to a specific binding member or analyte by a cleavable linker. In one embodiment, the acridinium compound is an acridinium-9-carboxamide. Methods for preparing acridinium 9-carboxamides are described in Mattingly, J. Biolumin. Chemilumin.6: 107-114 (1991); Adamczyk et al., J. Org. Chem.63: 5636-5639 (1998); Adamczyk et al.,Docket No. ABBTT-43595.601 Tetrahedron 55: 10899-10914 (1999); Adamczyk et al., Org. Lett.1: 779-781 (1999); Adamczyk et al., Bioconjugate Chem.11: 714-724 (2000); Mattingly et al., In Luminescence Biotechnology: Instruments and Applications; Dyke, K. V. Ed.; CRC Press: Boca Raton, pp.77–105 (2002); Adamczyk et al., Org. Lett.5: 3779-3782 (2003); and U.S. Patent Nos.5,468,646, 5,543,524 and 5,783,699 (each of which is incorporated herein by reference in its entirety for its teachings regarding same). Another example of an acridinium compound is an acridinium-9-carboxylate aryl ester. An example of an acridinium-9-carboxylate aryl ester of formula II is 10-methyl-9- (phenoxycarbonyl)acridinium fluorosulfonate (available from Cayman Chemical, Ann Arbor, MI). Methods for preparing acridinium 9-carboxylate aryl esters are described in McCapra et al., Photochem. Photobiol., 4: 1111-21 (1965); Razavi et al., Luminescence 15: 245-249 (2000); Razavi et al., Luminescence 15: 239-244 (2000); and U.S. Patent No.5,241,070 (each of which is incorporated herein by reference in its entirety for its teachings regarding same). Such acridinium-9-carboxylate aryl esters are efficient chemiluminescent indicators for hydrogen peroxide produced in the oxidation of an analyte by at least one oxidase in terms of the intensity of the signal and / or the rapidity of the signal. The course of the chemiluminescent emission for the acridinium-9-carboxylate aryl ester is completed rapidly, i.e., in under 1 second, while the acridinium-9-carboxamide chemiluminescent emission extends over 2 seconds. Acridinium-9-carboxylate aryl ester, however, loses its chemiluminescent properties in the presence of protein. Therefore, its use requires the absence of protein during signal generation and detection. Methods for separating or removing proteins in the sample are well-known to those skilled in the art and include, but are not limited to, ultrafiltration, extraction, precipitation, dialysis, chromatography, and / or digestion (see, e.g., Wells, High Throughput Bioanalytical Sample Preparation. Methods and Automation Strategies, Elsevier (2003)). The amount of protein removed or separated from the test sample can be about 40%, about 45%, about 50%, about 55%, about 60%, about 65%, about 70%, about 75%, about 80%, about 85%, about 90%, or about 95%. Further details regarding acridinium-9-carboxylate aryl ester and its use are set forth in U.S. Patent App. No.11 / 697,835, filed April 9, 2007. Acridinium-9-carboxylate aryl esters can be dissolved in any suitable solvent, such as degassed anhydrous N,N-dimethylformamide (DMF) or aqueous sodium cholate. In some aspects, the detectable label is a detectable label that can be attached to the surface of a microparticle or bead. The label can be attached to the surface of the microparticles, such as by using a linker and other techniques known in the art. Examples of detectable labels that can be attached to the surface of a microparticle or bead include fluorescent dyes, colormetric labels or radionuclides. “Microsampling device” as used herein refers to any device known in the art that is suitable for extracting capillary blood through the skin. It is understood that while a sample obtainedDocket No. ABBTT-43595.601 through the skin using a microsampling device will comprise predominantly capillary blood, the sample may also comprise a small amount or percentage of interstitial fluid. In some aspects, the microsampling device can comprise from about 0.1 mL to about 4 mL of capillary blood. In some other aspects, the device contains a plurality of microneedles, lancets or microlancets, blades or microblades, microscrews, or any combination thereof. In some aspects, the plurality of microneedles, lancets or microlancets, blades or microblades, microscrews, or any combination thereof can be rotating. In yet other aspects, the plurality of microneedles, lancets or microlancets, blades or microblades, microscrews, or any combination thereof are non-rotating. In some aspects, the microsampling device creates a vacuum and / or uses a stored vacuum to pull the skin into the device and / or activate the plurality of microneedles, lancets or microlancets, blades or microblades, microscrews, or any combination thereof to cut the skin. Examplary microsampling devices which can be used in the methods described herein include the TAP device available from YourBio Health, Inc. (Cambridge, MA) as well as the device described in U.S. Patent No.9,113,836, the contents of which are herein incorporated by reference, the Tasso+, Tasso-M20, and Tasso-ST devices available from Tasso, Inc. (Seattle, WA), the One Draw device available from Draw Bridge Health (San Diego, CA), PBS-1000 from PreciHealth (Neuchatel, Switzerland) or the Loop blood collection device available from Loop Medical (Lausanne, Switzerland). In other aspects, an example of a microsampling device includes a fingerstick device. In some aspects, the microsampling device can include a band-aid, bandage, or other suitable material which can be applied or dispensed to the area of the skin once the sample is obtained and / or the device is removed and / or detached from the skin. “Normalize” Normalize” or “normalizing” as used herein refers adjusting the amount of an analyte determined in a capillary blood sample obtained from a subject based on the amount of the same analyte in venous blood. In some aspects, for example, normalizing can involve multiplying a factor (e.g., correlation or conversion factor) by the amount of the analyte in the capillary blood sample. As used herein, the term “partition” refers to a volume of fluid (e.g. liquid or gas) that is a separated portion of a bulk volume. A bulk volume may be partitioned into any suitable number (e.g. 10<2 >...10<3 >...10<4 >...10<5 >...10<6 >...10<7>, etc.) of smaller volumes (i.e. partitions). Partitions may be separated by a physical barrier or by physical forces (e.g. surface tension, hydrophobic repulsion, etc.). Partitions generated from the larger volume may be substantially uniform in size (monodisperese) or may have non-uniform sizes (polydisperse). Partitions may be produced by any suitable manner (e.g. emulsion, microfluidics, microspray, etc.). Exemplary partitions are droplets. Partitions are referred to as “subvolumes” and “microvolumes.” “Point-of-care device” refers to a device used to provide medical diagnostic testing at or near the point-of-care (namely, typically, outside of a laboratory), at the time and place of patient care (such as in a hospital, physician’s office, urgent or other medical care facility, a patient’s home, aDocket No. ABBTT-43595.601 nursing home and / or a long-term care and / or hospice facility). Examples of point-of-care devices include those produced by Abbott Laboratories (Abbott Park, IL) (e.g., i-STAT and i-STAT Alinity, Universal Biosensors (Rowville, Australia) (see US 2006 / 0134713), Axis-Shield PoC AS (Oslo, Norway) and Clinical Lab Products (Los Angeles, USA). As used herein, the term “primer” refers to an oligonucleotide, whether occurring naturally as in a purified restriction digest or produced synthetically, that is capable of acting as a point of initiation of synthesis when placed under conditions in which synthesis of a primer extension product that is complementary to a nucleic acid strand is induced (e.g., in the presence of nucleotides and an inducing agent such as a biocatalyst (e.g., a DNA polymerase or the like) and at a suitable temperature and pH). The primer is typically single stranded for maximum efficiency in amplification, but may alternatively be double stranded. If double stranded, the primer is generally first treated to separate its strands before being used to prepare extension products. In some embodiments, the primer is an oligodeoxyribonucleotide. The primer is sufficiently long to prime the synthesis of extension products in the presence of the inducing agent. The exact lengths of the primers will depend on many factors, including temperature, source of primer and the use of the method. ““Quality control reagents” in the context of immunoassays and kits described herein, include, but are not limited to, calibrators, controls, and sensitivity panels. A “calibrator” or “standard” typically is used (e.g., one or more, such as a plurality) in order to establish calibration (standard) curves for interpolation of the amount of an analyte, such as an antibody or an analyte. Alternatively, a single calibrator, which is near a reference level or control level (e.g., “low”, “medium”, or “high” levels), can be used. Multiple calibrators (i.e., more than one calibrator or a varying amount of calibrator(s)) can be used in conjunction to comprise a “sensitivity panel.” As used herein, a “reaction vessel” refers to a holder or receiver, such as a container, receptacle, tube, a well (e.g., an array of wells), microwell, and / or cartridge, in or upon which an assay is performed. In some embodiments, the reaction vessel is a well or array of wells. In some embodiments, the well or array of wells has a hydrophilic surface. In other embodiments, the well or array of wells has a hydrophobic surface. In some embodiments, when the reaction vessel is a well or array of wells, each well is of sufficient size to accommodate at least one solid support (e.g., such as a bead, microparticle, tube etc.) and to detect one or more analytes of interest in the well(s). “Result” as used herein refers to an item of information obtained by performing an assay. In one aspect, a result is an amount of a biomarker in a test sample (e.g, blood sample). In another aspect, a result is identifying the presence of biomarker in a sample. A result can be visually displayed (e.g., as a readout). “Plasma separation device” as used herein, refers to an apparatus or device that can be used to separate components of whole blood (e.g., blood cells and platelets) from serum, plasma or serum and plasma using a separation system, such as, for example, at least one membrane, filter,Docket No. ABBTT-43595.601 synthetic paper (e.g., micropillar scaffolds) or any combination thereof. Separation of whole blood components can produce a fraction of plasma (“plasma fraction”) and a blood cell and platelet fraction. Any plasma separation device known to the skilled artisan can be used to separate components of whole blood. Examples of suitable plasma separation devices of use herein include, without limitation, an automated blood cell separator device, a capillary flow-driven microfluidic device (e.g., for point-of-care plasma separation), a centrifugal microfluidic disk (lab-on-a-disk) device using a separator gel, a centrifuge, a deterministic lateral displacement (DLD) microfluidic chip, a plasma separation card. “Specific binding” or “specifically binding” as used herein may refer to the interaction of an antibody, a protein, or a peptide with a second chemical species, wherein the interaction is dependent upon the presence of a particular structure (e.g., an antigenic determinant or epitope) on the chemical species; for example, an antibody recognizes and binds to a specific protein structure rather than to proteins generally. If an antibody is specific for epitope “A”, the presence of a molecule containing epitope A (or free, unlabeled A), in a reaction containing labeled “A” and the antibody, will reduce the amount of labeled A bound to the antibody. “Specific binding member” is a member of a specific binding pair. A specific binding pair comprises two different molecules, which specifically bind to each other through chemical or physical means. Therefore, in addition to antigen and antibody specific binding pairs of common immunoassays, other specific binding pairs can include biotin and avidin (or streptavidin), carbohydrates and lectins, complementary nucleotide sequences, effector and receptor molecules, cofactors and enzymes, enzymes and enzyme inhibitors, and the like. Furthermore, specific binding pairs can include members that are analogs of the original specific binding members, for example, an analyte-analog. Immunoreactive specific binding members include antigens, antigen fragments, and antibodies, including monoclonal and polyclonal antibodies as well as complexes and fragments thereof, whether isolated or recombinantly produced, and aptamers. “Statistically significant” as used herein refers to the likelihood that a relationship between two or more variables is caused by something other than random chance. Statistical hypothesis testing is used to determine whether the result of a data set is statistically significant. In statistical hypothesis testing, a statistically significant result is attained whenever the observed p-value of a test statistic is less than the significance level defined of the study. The p-value is the probability of obtaining results at least as extreme as those observed, given that the null hypothesis is true. Examples of statistical hypothesis analysis include Wilcoxon signed-rank test, t-test, Chi-Square or Fisher’s exact test. “Significant” as used herein refers to a change that has not been determined to be statistically significant (e.g., it may not have been subject to statistical hypothesis testing). “Subject” and “patient” as used herein interchangeably refers to any vertebrate, including, but not limited to, a mammal (e.g., cow, pig, camel, llama, horse, goat, rabbit, sheep, hamsters, guineaDocket No. ABBTT-43595.601 pig, cat, dog, rat, and mouse, a non-human primate (for example, a monkey, such as a cynomolgus or rhesus monkey, chimpanzee, etc.) and a human). In some embodiments, the subject may be a human or a non-human. In some embodiments, the subject is a human. The subject or patient may be undergoing other forms of treatment. As used herein, a “transfer tube” refers to a container or receptacle used to transfer a fluid (e.g., a blood sample) from one location to a second location (e.g., to a reaction vessel or from a plasma separation device). “Venous blood” as used herein refers to a blood sample that is obtained from the veins from a subject using a syringe, needle, or combination thereof, or any appropriate device. In some embodiments, a venous blood sample is obtained by a trained health clinician such as a physician, phlebotomist, nurse, laboratory technician, or combination thereof. In some embodiments, a venous blood sample is whole blood, serum or plasma. Unless otherwise defined herein, scientific and technical terms used in connection with the present disclosure shall have the meanings that are commonly understood by those of ordinary skill in the art. For example, any nomenclatures used in connection with, and techniques of, cell and tissue culture, molecular biology, immunology, microbiology, genetics and protein and nucleic acid chemistry and hybridization described herein are those that are well known and commonly used in the art. The meaning and scope of the terms should be clear; in the event, however of any latent ambiguity, definitions provided herein take precedent over any dictionary or extrinsic definition. Further, unless otherwise required by context, singular terms shall include pluralities and plural terms shall include the singular. 2. Method of calibrating a volume of a liquid in a sample and determining a volume of an aliquot of the sample An aspect of the present disclosure provides systems, methods and kits for determining a volume of a liquid in a sample, for example, to calibrate a volume of a liquid in a sample to accurately determine the volume of an aliquot of the sample. In one embodiment, the systems, methods and kits for calibrating a volume of a liquid in a sample to determine a volume of an aliquot of the sample can involve the steps of (a) dispersing a predetermined number of microparticles or beads per unit volume of a predetermined first volume of liquid in a first sample, thereby calibrating the first volume of liquid in the first sample based on the number of microparticles or beads per µL of liquid in the first sample; (b) aliquoting an indeterminate second volume of liquid from the first sample into a container to provide a second sample comprising a number of microparticles or beads per µL of liquid that is proportionate to the number of microparticles per µL of liquid in the first sample; and (c) determining the volume of liquid in the second sample based on the number of microparticles or beads in the second sample, wherein the volume of liquid in the second sample is equal to the volume obtained by multiplying theDocket No. ABBTT-43595.601 predetermined first volume of liquid in the first sample by the quotient of the number of microparticles in the second sample divided by the predetermined number of microparticles dispersed in the first sample. In another embodiment, the systems, methods and kits of the first aspect of the disclosure involve (a) dispersing a predetermined number of microparticles or beads per unit of a predetermined first volume of liquid into a first sample; (b) providing a second sample comprising an indeterminate second volume of liquid obtained from the predetermined first volume of liquid; and (c) determining the indeterminate second volume of liquid in the second sample based on the number of microparticles in the second sample. To determine the indeterminate second volume of liquid in the second sample (e.g., aliquot taken from the first sample), the number of microparticles or beads in the second sample can be counted. In some instances, it may be desirable to count the number of microparticles or beads remaining in the volume of liquid in the first sample. The number of microparticles in the first sample can also be counted before the second sample is provided (e.g., in a solution prior to being dispersed in the first sample, or after being dispersed in the first sample). The indeterminate second volume of liquid (e.g., in the aliquot) can be determined by the following Equation 1: Equation 1 Volume = (number of microparticles or beads counted in the second sample / predetermined number of microparticles or beads dispersed in the first sample) x (predetermined first volume of the liquid in the first sample). The skilled artisan will also appreciate that a dilution factor can also be taken into consideration when determining the volume of liquid in the second sample (e.g., aliquot). The second sample can be provided by aliquoting the indeterminate second volume of liquid from the first sample into a reaction vessel. The skilled artisan will appreciate that this step can be performed manually, for example, by a person in a clinic or in a laboratory setting. The aliquot of the indeterminate second volume of liquid can also be automated, for example, in a high-throughput or automated screening system described herein. It is to be understood that “first sample” and “second sample” are used as relative terms and the first sample is not necessarily limited to the first time that a sample is being processed. For example, a subject’s whole blood sample can be taken, subject to subsequent processing into a second sample of a smaller size, and then aliquots of the smaller size sample can be taken (e.g., into partitions of a microfluidic device or a series of reaction vessels (e.g., nanowells). In this context, the disclosure contemplates dispersing the predetermined number of microparticles into the first sample or the second sample. As such, both the first sample and the second sample can be referred to as the first sample and the second sample and the third sample can appropriately be referred to as the second sample.Docket No. ABBTT-43595.601 The systems and methods described herein increase the accuracy of determining the amount or presence of an analyte of interest (e.g., biomarker) in biological samples. In one embodiment, the present disclosure relates to improved methods for determining an amount (e.g., a quantitative measure) or the presence (e.g., a qualitative measure) of an analyte of interest (e.g., biomarker) in a venous blood sample obtained from a subject (e.g., a human subject). In another embodiment, the present disclosure relates to improved methods for determining an amount (e.g., a quantitative measure) or the presence (e.g., a qualitative measure) of an analyte of interest (e.g., biomarker) in a capillary blood sample obtained from a subject (e.g., a human subject). In yet another embodiment, the present disclosure relates to improved methods for obtaining a blood count (e.g., CBC). In yet another embodiment, the present disclosure relates to improved methods for obtaining a complete metabolic panel (CMP). In yet another embodiment, the present disclosure relates to improved methods for obtaining a lipid panel. In yet another embodiment, the disclosure relates to improved methods for obtaining a hemoglobin A1C (HbA1c) test. In another embodiment, the disclosure relates to improved methods for obtaining vitamin levels. In another embodiment, the disclosure relates to improved methods for measuring hormone levels. In another embodiment, the disclosure relates to improved methods for measuring mineral levels. 3. Method for obtaining an accurate blood count An aspect of the disclosure relates to using a volume calibrator of the disclosure to obtain an accurate complete blood count of a subject. Many clinical laboratory procedures require the collection and analysis of a patient's blood. A complete blood count (CBC), for example, provides detailed information about the cellular components of blood. The CBC is one of the most commonly ordered clinical blood tests and provides important information about the most common blood components. The CBC is used as a broad screening test to check for such disorders as anemia (decrease in red blood cells or hemoglobin), infection, leukemia, and many other diseases. It is a panel of tests that examine different parts of the blood. The CBC encompasses all or some of the following measurements: (1) a white blood cell (WBC) count expressed as a number of cells (thousands) per microliter of blood; (2) a white blood cell differential which classifies the WBCs into each type (neutrophils, lymphocytes, etc.) expressed as percentages of all WBCs; (3) a red blood cell (RBC) count expressed as millions per microliter; (4) a hemoglobin value expressed in grams per deciliter of blood; (4) hematocrit (HCT) or percentage of blood volume comprising red blood cells; (5) a platelet count expressed as thousands per microliter; (6) mean corpuscular volume (MCV)—the volume amount of hemoglobin inside the red blood cells expressed in femtoliters (10−15 liters); (7) mean corpuscular hemoglobin concentration (MCHC) or the percentage of all hemoglobin that is carried within the red blood cells; and (8) the red cell distribution width (RDW) which quantifies the variation in RBC size (RDW=100×standard deviation of RBC size / mean RBC size).Docket No. ABBTT-43595.601 In adults, blood tests such as the CBC require the collection of a representative sample of the patient's blood—obtained most often by venipuncture. In the venipuncture procedure, blood is drawn from a vein, usually from the inside of the elbow (antecubital) or the back of the hand. The puncture site is cleaned with antiseptic, and a tourniquet (usually an elastic band) or blood pressure cuff is placed around the upper arm to apply pressure and restrict blood flow through the vein which causes veins below the tourniquet to distend (fill with blood). Next, a needle is inserted into the vein, and the blood is collected in an air-tight vial or a syringe usually containing an additive specific to the type of blood analysis to be performed. While the needle is inserted, the tourniquet is removed to restore circulation. Once the blood has been collected, the needle is removed, and the puncture site is covered to stop any bleeding. For infants and young children, venipuncture is not recommended and instead an acral body skin site such as a fingertip or heel is cleansed with antiseptic and punctured with a sharp needle or blade—collectively referred to as a lancet. The blood is collected in a pipette (small tube), on a slide, onto a test strip, or into a small container all of which may contain an additive such as an anticoagulant depending on the subsequent analysis to be performed. After collection, bandage may be applied to the puncture site to avoid continued bleeding. In an aspect, the disclosure provides a method of obtaining an accurate blood count of a patient, the method comprising: (a) obtaining a predetermined volume of liquid comprising a whole blood sample taken from a patient; (b) dispersing a predetermined number of microparticles per µL of the predetermined volume of liquid in the whole blood sample, thereby calibrating the volume of liquid in the whole blood sample based on the number of microparticles per µL of liquid in the whole blood sample; (c) aliquoting an indeterminate volume of liquid from the whole blood sample into a container to provide an aliquoted sample comprising a number of microparticles per µL of liquid that is proportionate to the number of microparticles per µL of liquid in the whole blood sample; (d) determining the volume of liquid in the aliquoted sample based on the number of microparticles in the aliquoted sample, wherein the volume of liquid in the aliquoted sample is equal to the volume obtained by multiplying the predetermined volume of liquid in the whole blood sample by the quotient of the number of microparticles in the aliquoted sample divided by the predetermined number of microparticles dispersed in the whole blood sample; and (e) determining a blood count of the patient based on the volume of liquid determined in the aliquoted sample. The skilled practitioner can use the volume of liquid determined in the aliquoted sample to determine one or more blood cell counts as part of a CBC using the information shown in Table 2 below. Table 2Docket No. ABBTT-43595.601The method can be used to obtain the following measurements: (1) a WBC count expressed as a number of cells (thousands) per microliter of blood; (2) a WBC differential which classifies the WBCs into each type (neutrophils, lymphocytes, etc.) expressed as percentages of all WBCs; (3) a RBC count expressed as millions per microliter; (4) a hemoglobin value expressed in grams per deciliter of blood; (4) hematocrit (HCT) or percentage of blood volume comprising red blood cells; (5) a platelet count expressed as thousands per microliter; (6) mean corpuscular volume (MCV)—the volume amount of hemoglobin inside the red blood cells expressed in femtoliters (10−15 liters); (7) mean corpuscular hemoglobin concentration (MCHC) or the percentage of all hemoglobin that is carried within the red blood cells; and (8) the red cell distribution width (RDW) which quantifies the variation in RBC size (RDW=100×standard deviation of RBC size / mean RBC size). In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a CBC. In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a WBC count. In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a WBC differential count. In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a lymphocyte count (e.g., T&B). In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a neutrophil count. In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a basophil count. In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises an eosinophil count. In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a RBC count (e.g., erythrocytes). In some embodiments, the blood count determined based on the volume ofDocket No. ABBTT-43595.601 liquid determined in the aliquoted sample comprises a reticulocyte count. In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a hemoglobin count. In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a hematocrit percentage. In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a MCV. In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a MCHC. In some embodiments, the blood count determined based on the volume of liquid determined in the aliquoted sample comprises a RDW count. In some aspects, an additional reagent is added to the sample to prepare the sample for analysis, for example, an anticoagulant, to prevent cells in the sample from clotting and ensure a uniform distribution of the cells suspended in the volume of liquid. 4. Counting Methods Aspects of the disclosure involve counting microparticles or beads in small volumes of liquid (e.g., aliquots) to determine the volume of liquid in the aliquot. The disclosure contemplates the use of any counting instrument that is known to the skilled artisan. Examples include microscopy, optical based counters (e.g., fluorescence counters, optical absorbed counters, scattering counters, refractive index counters, etc.), electrical-based counters (e.g., resistance-based counters capacitance- based counters, inductance-based counters, impedance-based counters, etc.), photoacoustic-based counters, magnetic based counters, sensors (e.g., nanopore sensor), thermal-based counters, vision- or imaging-based automated counters. The type of counting instrument employed for any particular application depends on the type of microparticle used as a volume calibrator in accordance with the methods disclosed herein. For example, in instances in which the microparticle is a radionuclide (such as an alpha-, beta-, or gamma-emitting radionuclide), the counting instrument can be a Geiger counter or an analytical methods involving gas proportional detectors for alpha and beta-particle counting (GP), sodium iodide or high resolution germanium detectors for gamma detection and spectrometry, low- energy gamma- or X-ray detectors, kinetic phosphorimeter analysis, mass spectrometric analyses (e.g., inductively coupled plasma-mass spectrometry, thermal ionization mass spectrometry, accelerator mass spectrometry), and neutron activation. The skilled artisan will appreciate that the radionuclide used and instrument for its detection depend on the type of emitter of the radionuclide (e.g., whether it’s an alpha-, beta-, or gamma-emitter). In other instances, in which the microparticle is a colored or fluorescent microparticle (e.g., microsphere), the microparticles can be counted in an optical chamber or using an optical based counter.Docket No. ABBTT-43595.601 The disclosure also contemplates counting microparticles using an automated flow cytometry system to automatically generate microparticle counts, in addition to additional cytometric data (e.g., fluorescence emission spectra, light scatter, transmission spectra, Raman spectra, etc.), for a plurality of biological samples. The microparticles can be dispersed in samples disposed within respective wells of a multi-well sample plate or other multi-sample container. The methods described herein can employ such an automated flow cytometry system to accurately determine a volume of liquid in a multi-well sample plate, for example, the volume of an aliquot taken from a multi-well plate, or the volume of liquid aliquoted into a multi-well plate. An automated flow cytometry system may be used to generate, in an automated fashion, microparticle counts, in addition to additional cytometric data (e.g., fluorescence emission spectra, light scatter, transmission spectra, Raman spectra, etc.), for a plurality of biological samples. These samples can be disposed within respective wells of a multi-well sample plate or other multi-sample container. Use of such an automated flow cytometry system can significantly reduce the personnel costs of assessing biological samples, as well as increasing the consistency, with respect to timing, positioning, and volumetric parameters of the liquid in samples or other data generated when compared to manually presenting samples to a flow cytometry apparatus. Further, such an automated flow cytometry system can be configured to operate within an incubator or other controlled environment, removing the need to remove the samples from an incubator for assessment. Additionally, such an automated system can assess samples for extended periods of time and / or at inconvenient times (e.g., overnight), reducing technician costs and / or logistics. The technical advantages of a system operating in the manner described herein are related to the enhanced quality control of the screening platform. Aspects of the disclosure involving using resistive pulse sensing to count the predetermined number of microparticle remaining in a sample (e.g., an aliquot). In a microfluidics example (e.g., digital) beads can be counted in a cell mixture (e.g., whole blood sample) using a microfluidic chip having a magnet inserted along a channel, a signal amplification circuit, and a data acquisition device (e.g., LABVIEW®) (see e.g., Song et al. Automatic detecting and counting magnetic beads-labeled target cells from a suspension in a microfluidic chip. Electrophoresis 2019; 40: 897–905, which is incorporated herein by reference). The microfluidic chip may be divided functionally into a separation zone and a detection zone. As a volume of liquid in a sample containing a predetermined number microparticles (e.g., magnetic beads) dispersed therein flows through the device the magnet can separate the beads at the separation zone and the beads can be counted at the detection zone using a resistive pulse sensor. In some instances, the microparticles are counted using a sensor (e.g., nanopore sensor). In other instances, the microparticles can be counted using an imaging- or vision-based automated counter. For instance, cameras and software algorithms (machine learning) can be used to count theDocket No. ABBTT-43595.601 microparticles or beads. In still other instances, particle image velocimetry can be used to count the microparticles or beads. In certain instances, the optical-based counter comprises light scattering or light blocking microparticle or bead counting. 5. Microparticles The first aspect of the disclosure relates to systems, methods and kits comprising involving the use of microparticles as a volume calibrator to accurately determine a volume of liquid in an aliquot taken from a predetermined volume of a liquid sample in which analytes of interest (e.g., cells, proteins, nucleic acids, organic compounds, etc.) are suspended. Dispersing a predetermined number of microparticles per unit volume in the predetermined volume of liquid provides a ratio of microparticles per unit volume of the liquid that remains constant when the microparticles are dispersed with the analytes of interest suspended in the sample, enabling the accurate determination of the volume of the aliquot taken from the sample simply by counting the microparticles present in the aliquot and then using the ratio to calculate the volume of the aliquot. This method compensates for variable volumes due to loss of fluid during transfer. The present disclosure is not limited to any type of microparticles, however, it is preferrable that the microparticles are inert so that they remain in suspension dispersed in the sample volume of liquid during the aliquoting process to ensure that the ratio of microparticles per unit volume of predetermined liquid remains constant during the transfer. In some embodiments, the microparticles comprise inert microparticles. In some embodiments, the microparticles comprise monodisperse microparticles. In an embodiment, the microparticles comprise monodisperse microspheres. Suitable monodisperse microparticles exhibit high sphericity and tight particle size distribution, often with CV less than 10 percent and specification of greater than 95% of particles in the desired size range. In an embodiment, the microparticles comprise monodisperse silver-coated silica microspheres. In an exemplary embodiment, the microparticles comprise microspheres or beads. The skilled artisan will appreciate that the type of microparticle (e.g., microsphere) used for any particular application can vary, for example, depending on the instrument on which the aliquoting is performed and used to count the microparticles. Exemplary microspheres of use herein include, without limitation, glass microspheres, polymer microspheres, ceramic microspheres, metal microspheres, metal-coated microspheres, fluorescent microspheres, conductive microspheres, retroflective microspheres, phosphorescent microspheres, photocatalytic microspheres, paramagnetic microspheres. The predetermined number of microparticles to use for volume calibration in any particular sample is determined based on the predetermined volume of liquid in the sample to be aliquoted. Preferably, the predetermined number of microparticles per unit volume (e.g., number of microspheres or beads per µL of liquid in the sample) should be selected to minimize the error. TheDocket No. ABBTT-43595.601 predetermined number of microparticles per unit of the volume of liquid can be between 1 and 10000, 100 and 5000, 500 and 2000, 750 and 1500, 800 and 1400, 900 and 1100, or between 950 and 1050 microparticles per unit of the volume of liquid. In an embodiment, the predetermined number of microparticles dispersed in the sample comprises between about 1 and about 10000 microparticles per µL of liquid in the sample. In an embodiment, the predetermined number of microparticles dispersed in the sample comprises between about 100 and about 5000 microparticles per µL of liquid in the sample. In an embodiment, the predetermined number of microparticles dispersed in the sample comprises between about 500 and about 2000 microparticles per µL of liquid in the sample. In an embodiment, the predetermined number of microparticles dispersed in the sample comprises between about 750 and about 1500 microparticles per µL of liquid in the sample. In an embodiment, the predetermined number of microparticles dispersed in the sample comprises between about 800 and about 1400 microparticles per µL of liquid in the sample. In an embodiment, the predetermined number of microparticles dispersed in the sample comprises between about 900 and about 1100 microparticles per µL of liquid in the sample. In an embodiment, the predetermined number of microparticles dispersed in the sample comprises between about 950 and about 1050 microparticles per µL of liquid in the sample. In an embodiment, the predetermined number of microparticles dispersed in the sample comprises about 1000 microparticles per µL of liquid in the sample. The predetermined number of microparticles can be dispersed into the volume of liquid of a sample to be aliquoted for subsequent analysis in either liquid form or combined dry with the sample. In some embodiments, the predetermined number of microparticles are provided in the form of a liquid. The liquid containing the predetermined number of microparticles is not critical. Any liquid that does not disrupt the sample can be used. For example, in some aspects, the liquid is a buffer or water. In some aspects, the predetermined number of microparticles are in the form of a liquid suspension. In some aspects, the predetermined number of microparticles are in the form of a homogeneous liquid suspension. In some further aspects, the predetermined number of microparticles are in a liquid suspension and are in Brownian motion. In still further aspects, the predetermined number of microparticles are in a homogeneous liquid suspension in Brownian motion. In some embodiments, the predetermined number of microparticles (e.g., microspheres or beads) are provided in a container in dry form. In other embodiments, the predetermined number of microparticles (e.g., microspheres or beads) are provided in a container suspended in a solution (e.g., comprising a buffer, surfactant, anti-coagulant, or other reagent that is needed depending on the type of microparticle used and the analyte of interest). When dispersed in liquid form, it is preferable to that the volume of fluid comprising the predetermined number of microparticles be significantly less than the volume of liquid of the sample to be aliquoted. For example, the volume of fluid comprising the predetermined number of microparticles to be dispersed is 2-fold less, 2.5-fold less, 3-fold less, 3.5-fold less, 4-fold less, 4.5-fold less, 5-fold less, 5.5-fold less, 6-fold less, 6.5-fold less, 7-fold less, 7.5-fold less, 8-fold less, 8.5-fold less, 9-fold less, 9.5-fold less, 10-fold less, 11-fold less, 12-foldDocket No. ABBTT-43595.601 less, 13-fold less, 14-fold less, 15-fold less, 16-fold less, 17-fold less, 18-fold less, 19-fold less, 20- fold, 21-fold less, 22-fold less, 23-fold less, 24-fold less, or 25-fold less or more than the volume of liquid of the sample to be aliquoted. In an embodiment, the volume of fluid comprising the predetermined number of microspheres is between about 5-fold and 15-fold less, between 6-fold and 14-fold less, between 7-fold and 13-fold less, between 8-fold and 12-fold less, between 9-fold and 11- fold less. In an embodiment, the volume of fluid comprising the predetermined number of microspheres is between about 10-fold less than the volume of the sample to be aliquoted. 6. Samples Aspects of the disclosure involve calibrating a volume of a liquid in a sample to determine a volume of an aliquot of the sample, for example, to use the calibrator to accurately determine the volume of the aliquot using the calibrators that are present in the aliquot taken from the sample. The disclosure is not limited to the type of sample in which a volume calibrator (e.g., microparticles of the disclosure) can be used to accurately determine a volume of liquid (e.g., in an aliquot of the sample). The systems and methods of the disclosure involve transferring (e.g., aliquoting) an indeterminate volume of liquid from a predetermined volume of liquid in a first sample (e.g., whole blood obtained from a subject) into a second sample (e.g., container, such as a reaction vessel) to provide a second sample comprising an indeterminate volume of liquid that can be accurately determined using a microparticle of the disclosure (e.g., volume calibrator). The volume of liquid aliquoted from the first sample into the second sample can vary depending on the needs of the assay for which the sample is being taken (e.g., CBC, complete metabolic panel (CMP), lipids, biomarkers for a disease or condition, etc.). In certain aspects, the predetermined first volume of liquid is between about 1µL and about 10 mL, between about 2µL and about 9 mL, between about 3µL and about 8 mL, between about 4µL and about 7 mL, between about 5µL and about 6 mL, between about 6µL and about 5 mL, between about 7µL and about 4 mL, between about 8µL and about 3 mL, between about 9µL and about 2 mL, or between about 10 µL and about 1 mL. In some embodiments, the predetermined first volume of liquid is about 1 µL, 5 µL, 10 µL, 15 µL, 20 µL, 25 µL, 30 µL, 35 µL, 45 µL, 50 µL, 55 µL, 60 µL, 65 µL, 75 µL, 80 µL, 85 µL, 90 µL, 95 µL, 100 µL, 150 µL, 200 µL, 250 µL, 300 µL, 350 µL, 400 µL, 450 µL, 500 µL, 550 µL, 600 µL, 650 µL, 700 µL, 750 µL, 800 µL, 850 µL, 900 µL, 950 µL, or about 1 mL. In certain aspects, the indeterminate second volume of liquid (e.g., aliquoted from the first volume of the first sample) is between about 10 nL and about 100 µL, between about 20 nL and about 90 µL, between about 30 nL and about 80 µL, between about 40 nL and about 70 µL, between about 50 nL and about 60 µL, between about 60 nL and about 50 µL, between about 70 nL and about 40 µL, between about 80 nL and about 30 µL, between about 90 nL and about 20 µL, or between aboutDocket No. ABBTT-43595.601 100nL and about 10 µL. In some embodiments, the indeterminate second volume of liquid is about 1 µL, 2 µL, 3 µL, 4 µL, 5 µL, 6 µL, 7 µL, 8 µL, 9 µL, or about 10 µLIn other embodiments, the indeterminate second volume of liquid is about 50 nL, 100 nL, 150 nL, 200 nL, 250 nL, 300 nL, 350 nL, 400 nL, 450 nL, 500 nL, 550 nL, 600 nL, 650 nL, 700 nL, 750 nL, 800 nL, 850 nL, 900 nL, 950 nL, 100 nL, 150 nL, 200 nL, 250 nL, 300 nL, 350 nL, 400 nL, 450 nL, 500 nL, 550 nL, 600 nL, 650 nL, 700 nL, 750 nL, 800 nL, 850 nL, 900 nL, 950 nL, or about 1 µL. In some instances, the sample is diluted. In other instances, the sample is undiluted. In some aspects, the sample is obtained from a subject (e.g., human subject) and comprises a whole blood sample. In other aspects, the sample comprises a plasma sample. In other aspects, the sample comprises a serum sample. In other aspects, the sample comprises a urine sample. In still other aspects, the sample comprises a cerebrospinal fluid sample. In some aspects, the sample comprises or is suspected to comprise one or more nucleic acids capable of analysis by the methods. Preferably, the samples comprise nucleic acids (e.g., DNA, RNA, cDNAs, microRNA, mitochondrial DNA, etc.). Samples may be complex samples or mixed samples, which contain nucleic acids comprising multiple different nucleic acid sequences (e.g. host and pathogen nucleic acids; mutant and wild-type species; heterogeneous tumor. Samples may comprise nucleic acids from more than one source (e.g. different species, different subspecies, etc.), subject, and / or individual. In some aspects, the sample obtained from a subject (e.g., a human subject) comprises a venous blood sample, such as, whole blood, serum or plasma. In other aspects, the sample obtained from a subject (e.g., a human subject) is a capillary blood sample, such as, whole blood, serum or plasma. In some aspects, the capillary blood sample is whole blood. In other aspects, the capillary blood sample is serum. In yet other aspects, the capillary blood sample is plasma. The capillary blood sample can be obtained by extracting the sample through the skin (e.g., such as the fingers and / or toes, a hand, a foot (including the heel), an earlobe, a location on the arms and / or legs, chest, back, head, or any combinations thereof) of a subject. In some aspects, the capillary blood sample is extracted from the fingers or toes. In other aspects, the whole capillary blood sample is extracted from the arms or legs. In still other aspects, the capillary blood sample is obtained from the hands or feet. In still other aspects, the capillary blood sample is obtained from the chest or back. In yet other aspects, the capillary blood sample is obtained from an earlobe. In still other aspects, the capillary blood sample is obtained from the head. In some aspects, the capillary blood sample obtained from the subject is obtained without the use of a syringe, needle (e.g., 21-gauge needle, a butterfly needle, etc.), or any other suitable device, or any combination thereof which are typically used to draw blood (e.g., venous blood). Instead, the capillary blood sample is obtained using a self- or other-administered blood collection device. Examples of self-or other-administered blood collection devices include microsamplingDocket No. ABBTT-43595.601 devices. Example microsampling devices which can be used herein include the TAP device available from YourBio Health, Inc. (Cambridge, MA) as well as the device described in U.S. Patent No. 9,113,836, the contents of which are herein incorporated by reference, the Tasso+, Tasso-M20, and Tasso-ST devices available from Tasso, Inc. (Seattle, WA), the One Draw device available from Draw Bridge Health (San Diego, CA), PBS-1000 from PreciHealth (Neuchatel, Switzerland) or the Loop blood collection device available from Loop Medical (Lausanne, Switzerland). In other aspects, the capillary blood sample is obtained or collected from a subject in a decentralized setting. For example, the capillary blood sample can be obtained or collected from an urgent care clinic, a pharmacy, a grocery or other convenience store, a residence, a workplace, and / or a government office. In addition or alternatively, in still yet further aspects, the capillary blood sample is obtained from the subject by a user who is not trained in collecting blood (e.g., by someone other than a trained phlebotomist, a nurse, a medical assistant and / or physician). For example, the capillary blood sample can be obtained by the subject, a relative, friend, a co-worker, a coach, a pharmacist, and / or any other individual. In still yet further aspects, the capillary blood sample is obtained from a subject by a robot. In some aspects, the capillary blood sample or venous blood sample obtained from the subject is in an amount of less than about 4 mL. In some aspects, the capillary blood or venous blood sample obtained from the subject is less than about 3 mL. In some aspects, the capillary or venous blood sample obtained from the subject is less than about 2 mL. In some aspects, the amount of capillary or venous blood sample obtained from the subject is less than about 3.9 mL, about 3.8 mL, about 3.7 mL, about 3.6 mL, about 3.5 mL, about 3.4 mL, about 3.3 mL, about 3.2 mL, about 3.1 mL, about 3.0 mL, about 2.9 mL, about 2.8 mL, about 2.7 mL, about 2.6 mL, about 2.5 mL, about 2.4 mL, about 2.3 mL, about 2.2 mL, about 2.1 mL, about 2.0 ml, about 1.9 mL, about 1.8 mL, about 1.7 mL, about 1.6 mL, about 1.5 mL, about 1.4 mL, about 1.3 mL, about 1.2 mL, about 1.1 mL, about 1.0 mL, about 0.9 mL, about 0.8 mL, about 0.7 mL, about 0.6 mL, or about 0.5 mL. In some aspects, higher volumes of capillary or venous blood may be obtained when the sample collected is whole blood. In some aspects, the sample (e.g., capillary or venous blood sample) obtained from a subject is whole blood that is subjected to further or additional processing prior to determining the amount of an analyte (e.g., biomarker) in the sample. In some aspects, the sample is processed using centrifugation. In yet other aspects, the sample is processed using a plasma separation device which may comprise at least one filter, membrane and / or synthetic paper. The plasma separation device can separate whole blood into serum and / or plasma which can then be used in the methods described herein.Docket No. ABBTT-43595.601 In other aspects, the plasma separation device can be in fluid communication with or operably linked, coupled and / or removably coupled to a microsampling device as part of a system. In other aspects, the plasma separation device can be integrated into the microsampling device. In still yet other aspects, the plasma separation device can be in fluid communication with or operably linked, coupled and / or removably coupled to an aperture of a reaction vessel. For example, in some aspects, the reaction vessel is a cartridge such as those used in a point-of-care device. In still other aspects, the plasma separation device can be in fluid communication with or operably linked, coupled and / or removably coupled to a transfer tube. In these aspects, the transfer can be in fluid communication with or operably linked, coupled and / or removably coupled to a reaction vessel. In still further aspects, the plasma separation device can be integrated into the transfer tube. In yet other aspects, the transfer tube includes a cap or a stopper. Examples of assays that can be used to determine the amount of an analyte (e.g., biomarker) of interest in the sample include, without limitation, an immunoassay, such as an enzyme immunoassay (EIA), an enzyme linked immunosorbent assay (ELISA), a fluorescent immunoassay, a chemiluminescence Immunoassay (CLIA), a radioimmunoassay (RIA), a microparticle enzyme immunoassay (MEIA), a turbidimetric immunoassay, etc. In yet other aspects, the assay may be a clinical chemistry assay such as, for example, a photometry, a spectrophotometry, an absorbance, a fluorescence, a turbidimetry, a nephelometry, a potentiometry and / or an electrophoresis assay. In yet further aspects, the assay may be a combination of an immunoassay and a clinical chemistry assay. In still other aspects, the assay may be a single molecule detection assay. The assays may be performed sequentially, in any order. Generally, a reference level of the analyte (e.g., biomarker) can also be employed as a benchmark against which to assess results obtained upon assaying a test sample for the biomarker. Generally, in making such a comparison, the reference level of the biomarker is obtained by running or conducting a particular assay a sufficient number of times and under appropriate conditions such that a linkage or association of analyte presence, amount or concentration with a particular stage or endpoint of an illness, disease and / or condition or with particular indicia can be made. Typically, the reference level of the biomarker is obtained with assays of reference subjects (or populations of subjects). The biomarker measured can include fragments thereof, degradation products thereof, and / or enzymatic cleavage products thereof. In certain aspects, the reference level may be correlated with control subjects (e.g., human subjects) that do not have the illness, condition, or disorder. In yet further aspects, once the amount of analyte (e.g., biomarker) is determined using the methods described herein, a result is obtained. This result can be further processed. Specifically, this further processing can involve selecting a conversion factor for comparing the amount of analyte (e.g.,Docket No. ABBTT-43595.601 biomarker) in the capillary whole blood or plasma sample with the amount of the same analyte in venous whole blood or plasma. Once the conversion factor is selected, the processing further involves normalizing the amount of analyte in the capillary whole blood or plasma sample with the amount of the same analyte from venous whole blood or plasma by applying the conversion factor to the amount of analyte in the sample. For example, the amount of the analyte in the capillary whole blood or plasma sample can be multiplied by the conversion factor to provide the normalized amount of the analyte in the sample. In some aspects, the processing of the amount of the analyte (e.g., biomarker) can be by a processing system which comprises a computer processor and a non-transitory computer memory comprising one or more computer programs. In some embodiments, when a capillary sample is used, the one or more computer programs can be used in conjunction with said computer process to select a conversion factor for comparing the amount of the biomarker in the capillary whole blood or plasma sample with the amount of the same analyte in venous whole blood or plasma and normalize the amount of analyte in the sample with the amount of the same analyte from venous whole blood or plasma by applying the selected conversion factor to the amount analyte in the sample. This result or processed result can be communicated (e.g., reported) for further analysis, interpretation, processing and / or display. The result can be communicated (e.g., reported) by a computer, in a document and / or spreadsheet, on a mobile device (e.g., a smart phone), on a website, in an e-mail, or any combination thereof. In some aspects, the result can displayed not as a number, but as a visual signal (e.g., a line or bar) that can be read / interpreted by a reader or the naked eye. In some aspects, the result determined in the sample is communicated by being displayed, such as on an instrument. In further aspects, the result is displayed as indicating that the amount of analyte in a subject is elevated, are not elevated, or that the assay(s) for the analyte should be repeated. Suitable instruments for use in the methods described herein include a higher throughput assay analyzer (e.g., the ARCHITECT platform marketed by Abbott Laboratories) or a point-of-care device (e.g., i-STAT and i-STAT Alinity devices marketed by Abbott Laboratories) that may contain a user interface that can display the determination. In some aspects, the instrument contains software to execute one or more tasks, including the performance of the methods and algorithms described herein. In some aspects, the instrument contains software to automatically determine the next appropriate step in a methods and algorithms as described herein. For example, the instrument may contain software that determines the amount or presence of an analyte of interest. The software may display this determination, such as on a graphical user interface. In some aspects, the instrument stores software that instructs a processor to execute a given task. In some aspects, the software stores machine readable instructions that instruct a processor toDocket No. ABBTT-43595.601 execute a given task. The machine-readable instructions may be one or more executable programs or portion(s) of an executable program for execution by a computer. The programs may be embodied in software stored on a non-transitory computer readable storage medium such as a CD-ROM, a floppy disk, a hard drive, a DVD, a Blu-ray disk, or a memory associated with the processors. Alternatively, the entire programs and / or parts thereof could alternatively be executed by a device other than the processors and / or embodied in firmware or dedicated hardware. Additionally or alternatively, processes may be implemented by one or more hardware circuits (e.g., discrete and / or integrated analog and / or digital circuitry, an FPGA, an ASIC, a comparator, an operational-amplifier (op-amp), a logic circuit, etc.) structured to perform the corresponding operation without executing software or firmware. The machine-readable instructions may be stored in one or more of a compressed format, an encrypted format, a fragmented format, a compiled format, an executable format, a packaged format, etc. Machine readable instructions as described herein may be stored as data (e.g., portions of instructions, code, representations of code, etc.) that may be utilized to create, manufacture, and / or produce machine executable instructions. For example, the machine-readable instructions may be fragmented and stored on one or more storage devices and / or computing devices (e.g., servers). The machine-readable instructions may require one or more of installation, modification, adaptation, updating, combining, supplementing, configuring, decryption, decompression, unpacking, distribution, reassignment, compilation, etc. in order to make them directly readable, interpretable, and / or executable by a computing device and / or other machine. For example, the machine-readable instructions may be stored in multiple parts, which are individually compressed, encrypted, and stored on separate computing devices, wherein the parts when decrypted, decompressed, and combined form a set of executable instructions that implement a program such as that described herein. In another example, the machine-readable instructions may be stored in a state in which they may be read by a computer, but require addition of a library (e.g., a dynamic link library (DLL)), a software development kit (SDK), an application programming interface (API), etc. in order to execute the instructions on a particular computing device or other device. In another example, the machine-readable instructions may need to be configured (e.g., settings stored, data input, network addresses recorded, etc.) before the machine-readable instructions and / or the corresponding program(s) can be executed in whole or in part. Thus, the disclosed machine-readable instructions and / or corresponding program(s) are intended to encompass such machine-readable instructions and / or program(s) regardless of the particular format or state of the machine-readable instructions and / or program(s) when stored or otherwise at rest or in transit. The machine-readable instructions described herein can be represented by any past, present, or future instruction language, scripting language, programming language, etc. For example, the machine-readable instructions may be represented using any of the following languages: C, C++,Docket No. ABBTT-43595.601 Java, C#, Perl, Python, JavaScript, HyperText Markup Language (HTML), Structured Query Language (SQL), Swift, etc. The machine readable instructions may be stored on a non-transitory computer and / or machine readable medium such as a hard disk drive, a flash memory, a read-only memory, a compact disk, a digital versatile disk, a cache, a random-access memory and / or any other storage device or storage disk in which information is stored for any duration (e.g., for extended time periods, permanently, for brief instances, for temporarily buffering, and / or for caching of the information). As used herein, the term non-transitory computer readable medium is expressly defined to include any type of computer readable storage device and / or storage disk and to exclude propagating signals and to exclude transmission media. It may be desirable to include a control sample. The control sample may be analyzed concurrently with the sample from the subject as described above. The results obtained from the subject sample can be compared to the results obtained from the control sample. Standard curves may be provided, with which assay results for the sample may be compared. Such standard curves present levels of marker as a function of assay units, i.e., fluorescent signal intensity, if a fluorescent label is used. Using samples taken from multiple donors, standard curves can be provided for reference levels of an analyte (e.g., biomarker) in normal healthy subjects or tissue, as well as for “at-risk” levels of the analytes in tissue taken from donors, who may have one or more clinically relevant characteristics. 7. Digital microfluidics In certain embodiments, the methods described herein can be performed using digital microfluidics. For example, the methods described herein can be performed on or in one or more chips (e.g., lab-on-chip or biochip), cartridges, or combinations thereof. In certain embodiments, the methods described herein are performed using a digital microfluidics device (DMF). In certain embodiments, methods described herein are performed using a surface acoustic wave-based microfluidics device (SAW). In other embodiments, the methods described herein are performed using a micro-total analysis system or a biomedical (or biological) microelectromechanical system (BIOMEMs). In still other embodiments, the methods described herein are performed using an integrated DMF and analyte detection device. In certain embodiments, methods described herein are performed using an integrated surface acoustic wave based microfluidic device and analyte detection device. In certain embodiments, methods described herein are performed using a robotics based assay processing unit. For example, in some embodiments, methods such as those described in U.S. Patent Application Publication No.2018 / 0095067, the contents of which are herein incorporated by reference, can be used. Specifically, in some embodiments, the methods involve measuring an analyte of interest in a liquid droplet (wherein the analyte of interest is from a test or biological sample). TheDocket No. ABBTT-43595.601 method includes providing a first liquid droplet containing an analyte of interest, providing a second liquid droplet containing at least one solid support (such as, for example, a magnetic solid support (such as a bead or microparticle)) which contains a specific binding member that binds to the analyte of interest, using energy to exert a force to manipulate the first liquid droplet (which contains the analyte of interest) with the second liquid (containing the at least one solid support) to create a mixture, moving all or at least a portion of the mixture to an array of wells (where one or more wells of the array are of sufficient size to accommodate the at least one solid support), adding at least one detectable label to the mixture before, after or both before or after moving a portion of the mixture to the array of wells and measuring the analyte of interest in the wells. In certain embodiments, the droplet comprises a predetermined number of microparticles dispersed within it to calibrate a volume of fluid in the droplet. In certain embodiments, the detectable label is used to detect a specific analyte of interest (e.g., using a detection agent described herein to detect an antigen of interest). In certain embodiments, “using energy to exert a force to manipulate the first liquid droplet with the second liquid droplet” refers to the use of non-mechanical forces (namely, for example, energy created without the use of pumps and / or valves) to provide or exert a force that manipulates (such as merges or combines) at least the first and second liquid droplets (and optionally, additional droplets) into a mixture. Example of non-mechanical forces that can be used in the methods described herein include electric actuation force (such as droplet actuation, electrophoresis, electrowetting, dielectrophoresis, electrostatic actuation, electric field mediated, electrode mediated, capillary force, chromatography, centrifugation or aspiration) and / or acoustic force (such as surface acoustic wave (or “SAW”). In certain embodiments, the electric actuation force generated is an alternating current. For example, the alternating current can have a root mean squared (rms) voltage of 10 V, 15 V, 20 V, 25 V, 30 V, 35V or more. For example, such alternating current can have a rms voltage of 10 V or more, 15 V or more, 20 V or more, 25 V or more, 30 V or more or 35 V or more. Alternatively, the alternating current can have a frequency in a radio frequency range. In certain embodiments, if magnetic solid supports are used, an electric actuation force and a magnetic field can be applied and applied from opposition directions, relative to the at least a portion of the mixture. In certain other embodiments, the mixture is mixed by moving it: back and forth, in a circular pattern or by splitting it into two or more sub-mixtures and then merging the sub- mixtures. In certain other embodiments, an electric actuation force can be generated using a series or plurality of electrodes (namely, at least two or more, at least three or more, at least four or more, at least five or more, at least six or more, at least seven or more, at least eight or more, at least nine or more, at least ten or more, at least eleven or more, at least twelve or more, at least thirteen or more, at least fourteen or more, at least fifteen or more, etc.) to move the mixture to the array of wells in order to seal the wells (which are loaded with at least one solid support).Docket No. ABBTT-43595.601 In certain embodiments, the moving of all or at least a portion of the mixture to an array of wells results in the loading (filling and / or placement) of the at least one solid support into the array of reactions vessels (e.g., wells). In some embodiments, the mixture that is moved comprises a liquid sample in which a predetermined number of microparticles is dispersed to calibrate the volume of fluid in the sample as at least a portion of the mixture is moved to the array of wells. In certain embodiments, when wells are used, a magnetic field is used to facilitate movement of the mixture and thus, at least one solid support, into one or more wells of the array. In certain embodiments, after the at least one solid supports are loaded into the wells, any solid supports that are not loaded into a well can be removed using routine techniques known in the art. For example, such removing can involve generating an electric actuation force (such as that described previously herein) with a series or plurality of electrodes to move a fluid droplet (such as a polarizable fluid droplet) to the array of wells to move at least a portion of the mixture to a distance (the length of which is not critical) from the array of wells. In certain embodiments, an aqueous washing liquid can be used to remove the solid supports not bound to any analyte of interest. In such embodiments, the removal involves generating an electric actuation force with a series or plurality of electrodes to move an aqueous wash (or washing) droplet (a third droplet) across the array of wells. The amount and type of aqueous liquid used for said washing is not critical. In certain embodiments, the mixture in the method is an aqueous liquid. In other embodiments, the mixture is an immiscible liquid. In other embodiments, the liquid droplet is a hydrophobic liquid droplet. In other embodiments, the liquid droplet is a hydrophilic liquid droplet. In certain embodiments, the array of wells used in the method have a hydrophobic surface. In other embodiments, the array of wells has a hydrophilic surface. In certain embodiments, the first liquid droplet used in the method is a polarizable liquid. In certain embodiments, the second liquid droplet used in the method is a polarizable liquid. In certain embodiments, the first and second liquid droplets used in the method are polarizable liquids. In certain embodiments, the mixture is a polarizable liquid. In certain embodiments one or more of the first droplet, second droplet and mixture is a polarizable liquid. In some embodiments, the aqueous liquid comprises a predetermined number of microparticles dispersed within it to calibrate the volume of the liquid. In some embodiments, the immiscible liquid comprises a predetermined number of microparticles dispersed within it to calibrate the volume of the liquid. In some embodiments, the hydrophobic liquid droplet comprises a predetermined number of microparticles dispersed within it to calibrate the volume of the liquid together with a surfactant so that microparticles remain in suspension inside the droplet. In some embodiments, the hydrophilic liquid droplet comprises a predetermined number of microparticles dispersed within it to calibrate the volume of the liquid. In some embodiments, the array of wells comprises a predetermined number of microparticles dispersed within it to calibrate the volume of theDocket No. ABBTT-43595.601 liquid within each well, for example, so that as volumes of liquid move from well to well the volume of liquid can be used by counting the microparticles in each successive well. In certain embodiments, the at least one solid support comprises at least one binding member that specifically binds to the analyte of interest. In certain embodiments, the detectable label is added to the mixture before moving at least a portion of the mixture to the array of wells. In some embodiments, the detectable label is specific for detecting the analyte of interest (e.g., antigen). In certain other embodiments, the detectable label is added to the mixture after the moving of at least a portion of the analyte of interest to the array of wells. The detectable label can be added together with microparticles to calibrate a volume of liquid after moving at least the portion of the analyte of interest to the array of wells. In certain embodiments, the method further comprises positioning the at least a portion of the mixture over the array of wells, using a capillary element configured to facilitate movement of the mixture to the array of wells. In certain embodiments, the measuring first involves determining the total number of solid supports in a well, of the array (“total solid support number”). Next, the number of solid supports in the wells of the array that contain the detectable label are determined, such as, for example, determining the intensity of the signal produced by the detectable label (“positives”). The positives are subtracted from the total solid support number to provide the number of solid supports in the array of wells that do not contain a detectable label or are not detected (“negatives”). Then, the ratio of positives to negatives in the array of wells can be determined and then compared to a calibration curve. Alternatively, digital quantitation using the Poission equation P(x; μ) as shown below: P(x;μ)=(e−μ)(μx) / x! where: e: A is a constant equal to approximately 2.71828, μ: means number of successes that occur in a specified region, and x: is the tactual number of successes that occur in a specified region. Provided herein are methods for measuring or detecting an analyte present in a biological sample. The method includes contacting the sample with a first binding member, wherein the first binding member is immobilized on a solid support and wherein the first binding member specifically binds to the analyte; contacting the analyte with a second binding member, wherein the second binding member specifically binds to the analyte and wherein the second binding member includes a cleavable detectable label (or tag) attached thereto; removing second binding member not bound to the analyte bound to the first binding member; cleaving the detectable label attached to the second binding member that is bound to the analyte bound to the first binding member; translocating the cleaved detectable label through or across one or more nanopores in a layer; detecting or measuring detectable labels translocating through the layer; and assessing the detectable labels translocating through the layer, wherein measuring the number of detectable labels translocating through the layerDocket No. ABBTT-43595.601 measures the amount of analyte present in the sample, or wherein detecting labels translocating through the layer detects that the analyte is present in the sample. In some embodiments, measuring the labels translocating through the layer is assessed, wherein the number of labels translocating through the layer measures the amount of analyte present in the sample. In some embodiments, detecting the labels translocating through the layer is assessed, wherein detecting labels translocating through the layer detects that the analyte is present in the sample. In still yet other embodiments, each detectable label or tag, such as an aptamer, translocating through the layer is a translocation event. Measuring the number of translocation events measures the amount of analyte present in the sample. In some embodiments, the amount of analyte present in the sample can be determined by counting the number of translocation events during a set period of time and correlating the number of translocation events to a control. The standard curve can be determined by measuring the number of translocation events for control concentrations of analyte during a set period of time. In some embodiments, the amount of analyte present in the sample can be determined by measuring the amount of time for a set number of translocation events to occur and correlating to a control. The standard curve can be determined by measuring the time it takes for a set number of translocation events to occur for control concentrations of analyte. In some embodiments, the amount of analyte present in the sample can be determined by measuring the average time between translocation events to occur and correlating to a control. The standard curve can be determined by measuring the average time between translocation events to occur for control concentrations of analyte. In some embodiments, the control can be a reference standard comprising a calibration curve, standard addition, or digital polymerase chain reaction. For example, in some embodiments, the methods may include contacting the sample with a first binding member (“binding members” also referred to herein as “specific binding members,”), where the first binding member is immobilized on a solid support and where the first binding member specifically binds to the analyte; contacting the analyte with a second binding member, which second binding member specifically binds to the analyte and which second binding member includes a cleavable detectable label attached thereto; removing second binding member not bound to the analyte bound to the first binding member; cleaving the detectable labels attached to the second binding member that is bound to the analyte bound to the first binding member; translocating the detectable label through nanopores in a layer; determining the number of detectable labels translocating through the layer; determining concentration of the analyte in the sample based on the number of detectable labels translocating through the layer. In certain embodiments, the concentration of the analyte may be determined by counting the number of detectable labels translocating through the layer per unit time. In other embodiments, the concentration of the analyte may be determined by determining the time at which the number of detectable labels translocating through the layer reaches a threshold.Docket No. ABBTT-43595.601 In certain embodiments, the methods of the present disclosure are carried out using a device in which a digital microfluidics module is integrated with an analyte detection device, such as those known in the art. In certain embodiments, the digital integrated microfluidics module and the analyte detection device may be reversibly integrated. For example, the two modules may be combined physically to form the integrated device and which device could then be separated into the individual modules. In certain embodiments, the methods of the present disclosure are carried out using a disposable cartridge that includes a microfluidics module with a built-in analyte detection device. The moving of the droplets in the integrated microfluidic and analyte detection device may be carried out using electrical force (e.g., electrowetting, dielectrophoresis, electrode-mediated, opto- electrowetting, electric-field mediated, and electrostatic actuation) pressure, surface acoustic waves and the like. The force used for moving the droplets may be determined based on the specifics of the device using techniques known in the art. For example, in some embodiments, the methods include generating a droplet of the sample and combining the droplet of the sample with a droplet containing the first binding member to generate a single droplet. In this example, the sample or the droplet can comprise a predetermined number of microparticles dispersed within to calibrate a volume of fluid of the sample or the droplet before the droplet is combined. The first binding member may be immobilized on a solid substrate, such as, a bead (e.g., a magnetic bead). The single droplet may be incubated for a time sufficient to allow binding of the first binding member to an analyte present in the sample droplet. Optionally, the single droplet may be agitated to facilitate mixing of the sample with the first binding member. Mixing may be achieved by moving the single droplet back and forth, moving the single droplet around over a plurality of electrodes, splitting a droplet and then merging the droplets, or using SAWs, and the like. Next, the single droplet may be subjected to a magnetic force to retain the beads at a location in the device while the droplet may be moved away and replaced with a droplet containing a second binding member. An optional wash step may be performed, prior to adding the second binding member, by moving a droplet of wash buffer to the location at which the beads are retained using the magnetic force. After a period of time sufficient for the second binding member to bind the analyte bound to the first binding member, the droplet containing the second binding member may be moved away while the beads are retained at the first location. The beads may be washed using a droplet of wash buffer followed by contacting the beads with a droplet containing a cleavage reagent to cleave the tag attached to the second binding member. In embodiments where the tag is attached to the second binding member via a photocleavable linker, the beads may be exposed to light of the appropriate wavelength to cleave the linker. In certain cases, the beads may be exposed to a droplet of buffer prior to cleavage of the photocleavable linker. Optionally, after the washing step to remove any unbound second binding member, a droplet containing buffer may be left covering the beads, theDocket No. ABBTT-43595.601 magnetic force retaining the beads at the first location may be removed and the buffer droplet containing the beads may be moved to a second location at which the photocleavage may be carried out. The droplet containing the cleaved tags may then be moved to the nanopore device or the nanopore module portion of the integrated device. In embodiments using aptamer as the second binding member, after the washing step to remove any unbound aptamer, a droplet containing buffer may be left covering the beads, the magnetic force retaining the beads at the first location may be removed and the buffer droplet containing the beads may be moved to a second location at which the dissociation of the aptamer may be carried out. In other embodiments, after the washing step, the beads may be exposed to a droplet of a reagent for dissociating aptamer bound to the analyte. A droplet containing the dissociated aptamer may be moved to the nanopore while the beads may be retained in place using a magnet. The droplet containing the dissociated aptamer may be moved to the nanopore device or the nanopore module portion of the integrated device. In some embodiments, the first binding member may be immobilized on a surface of the first or the second substrate at a location in the gap / space. The step of contacting a sample with the first binding member may include moving a droplet of the sample to the location in the gap / space at which the first binding member is immobilized. The subsequent steps may be substantially similar to those described above for first binding member immobilized on magnetic beads. After the cleaving / dissociating step, the droplet containing the cleaved tag(s) / dissociated aptamer(s) may be moved to the nanopore device or the nanopore module of the integrated device. As noted above, the droplet(s) may be moved using a liquid transfer system, such as a pipette. In certain cases, the microfluidic module may be fluidically connected to the nanopore module. Fluidic connection may be achieved by connecting the microfluidics module to the nanopore module via a channel or by placing the nanopore module within the microfluidics module, either reversibly or during the manufacturing process of the integrated device. Such devices are further described in the following section. In the above embodiments, optionally, after the combining, a droplet may be manipulated (e.g., moved back and forth, moved in a circular direction, oscillated, split / merged, exposed to SAW, etc.) to facilitate mixing of the sample with the assay reagents, such as, the first binding member, second binding member, etc. The moving of the droplets in the integrated microfluidics nanopore device may be carried out using electrical force (e.g., electrowetting, dielectrophoresis, electrode-mediated, opto- electrowetting, electric-field mediated, and electrostatic actuation) pressure, surface acoustic waves and the like. The force used for moving the droplets may be determined based on the specifics of the device and can be determined using techniques known in the art.Docket No. ABBTT-43595.601 8. Amplification In some embodiments, the present disclosure provides compositions and methods for the amplification of target nucleic acids (e.g. DNA, RNA, etc.). In some embodiments, amplification is performed on a sample that has been divided into partitions (e.g. droplets). In some embodiments, an amplification reaction is carried out within each partition. In some embodiments, a predetermined number of microparticles is dispersed in the sample to calibrate the volume of liquid in the sample before it is partitioned (e.g., into droplets). The microparticles can be counted before being dispersed in the sample. The microparticles can be used to determine the volume of liquid in the droplets by counting the microparticles in each droplet using a counting instrument described herein and using Equation 1. The microparticles in any portion remaining of the original sample can also be counted to improve the accuracy of the method. In some embodiments, amplification is performed on a sample that has been aliquoted, for example, into a container or reaction vessel (e.g., an aliquot). In some embodiments, a predetermined number of microparticles is dispersed in the sample to calibrate the volume of fluid in the sample before it is aliquoted into the container or reaction vessel (e.g., aliquot). In some embodiments, the sample is aliquoted into a transfer tube. The microparticles can be counted before being dispersed in the sample. The microparticles can be used to determine the volume of liquid in the aliquots by counting the microparticles in each container or reaction vessel (e.g., aliquot) using a counting instrument described herein and using Equation 1. The microparticles in any portion remaining of the original sample can also be counted to improve the accuracy of the method. In some embodiments, a partition, container, or reaction vessel (e.g., aliquot) contains all the reagents necessary for nucleic acid amplification (e.g. primers, polymerase, deoxynucleotides, template (e.g. target, internal standard, etc.)). In some embodiments, amplification is performed using digital PCR (dPCR) methods. In some other embodiments, the amplification is performed using dPCR methods where the number of subvolumes containing 0 target is nonexistent or is too low to apply Poisson statistics. In some embodiments, the present disclosure provides compositions (e.g. primers, buffers, salts, nucleic acid targets, etc.) and methods for the amplification of nucleic acid (e.g. dPCR, digital droplet amplification, PCR amplification, partitioned amplification, cdPCR, dPCR LATE, combinations thereof, etc.). In some embodiments, an amplification reaction is any reaction in which nucleic acid replication occurs repeatedly over time to form multiple copies of at least one segment of a template or target nucleic acid molecule (e.g. DNA, RNA). In some embodiments, amplification reaction is achieved by repeated thermal cycling. In some embodiments, amplification reaction is achieved isothermally. In some embodiments, amplification generates an exponential or linear increase in the number of copies of the template nucleic acid. Amplifications may produce in excess of a 1,000-fold increase in template copy-number and / or target-detection signal.Docket No. ABBTT-43595.601 Exemplary amplification reactions include, but are not limited to, the polymerase chain reaction (PCR) or ligase chain reaction (LCR), each of which is driven by thermal cycling. Amplifications used in method or assays of the present disclosure may be performed in bulk and / or partitioned volumes (e.g. droplets, reactive vessels, etc.). Alternative amplification reactions, which may be performed isothermally, also find use herein, such as branched-probe DNA assays, cascade- RCA, helicase-dependent amplification, loop-mediated isothermal amplification (LAMP), nucleic acid-based amplification (NASBA), nicking enzyme amplification reaction (NEAR), PAN-AC, Q- beta replicase amplification, rolling circle replication (RCA), self-sustaining sequence replication, strand-displacement amplification, and the like. Amplification may be performed with any suitable reagents (e.g. template nucleic acid (e.g. DNA or RNA), primers, probes, buffers, replication catalyzing enzyme (e.g. DNA polymerase, RNA polymerase), nucleotides, salts (e.g. MgCl2), etc. In some embodiments, an amplification mixture includes any combination of at least one primer or primer pair, at least one probe, at least one replication enzyme (e.g., at least one polymerase, such as at least one DNA and / or RNA polymerase), and deoxynucleotide (and / or nucleotide) triphosphates (dNTPs and / or NTPs), etc. In some embodiments, the present disclosure utilizes nucleic acid amplification that relies on alternating cycles of heating and cooling (i.e., thermal cycling) to achieve successive rounds of replication (e.g., PCR). In some embodiments, PCR is used to amplify target nucleic acids (e.g. partitioned targets). PCR may be performed by thermal cycling between two or more temperature set points, such as a higher melting (denaturation) temperature and a lower annealing / extension temperature, or among three or more temperature set points, such as a higher melting temperature, a lower annealing temperature, and an intermediate extension temperature, among others. PCR may be performed with a thermostable polymerase, such as Taq DNA polymerase (e.g., wild-type enzyme, a Stoffel fragment, FastStart polymerase, etc.), Pyrococcus furiosus (Pfu) DNA polymerase, S-Tbr polymerase, Thermus thermophilus (Tth) polymerase, Vent polymerase, or a combination thereof, among others. Typical PCR methods produce an exponential increase in the amount of a product amplicon over successive cycles, although linear PCR methods also find use in the present disclosure. Any suitable PCR methodology, combination of PCR methodologies, or combination of amplification techniques may be utilized in the partitioned methods (e.g. droplet-based detection, separation, and / or sequencing of target nucleic acids, reaction vessels, etc.) disclosed herein, such as cdPCR, dPCR LATE, allele-specific PCR, assembly PCR, asymmetric PCR, digital PCR, endpoint PCR, hot-start PCR, in situ PCR, intersequence-specific PCR, inverse PCR, linear after exponential PCR, ligation-mediated PCR, methylation-specific PCR, mini-primer PCR, multiplex ligation- dependent probe amplification, multiplex PCR, nested PCR, overlap-extension PCR, polymerase cycling assembly, qualitative PCR, quantitative PCR, real-time PCR, RT-PCR, single-cell PCR, solid- phase PCR, thermal asymmetric interlaced PCR, touchdown PCR, or universal fast walking PCR, etc.Docket No. ABBTT-43595.601 In some embodiments, the present disclosure provides dPCR methods (See, e.g., Ramakrishnan, American Biotechnology Laboratory 27(8), 11-13 (2009); herein incorporated by reference in its entirety). In some embodiments, PCR is performed on portions of a sample (e.g. partitions) to determine the presence or absence, concentration, and / or copy number of a nucleic acid target in the sample, based on how many of the sample portions support amplification of the target. In some embodiments, PCR is performed on portions of a sample (e.g. partitions or reaction vessels) to determine the presence or absence, concentration, and / or copy number of a nucleic acid target in the sample, based on quantitation of sub-volume nucleic acid concentration determined by methods described herein (e.g., cdPCR, dPCR LATE). In some embodiments, PCR is performed in partitions (e.g., droplets) or reaction vessels comprising microparticles of the disclosure to calibrate the volume of fluid in the droplets or reaction vessels. In some embodiments, PCR is performed on portions of a sample (e.g. partitions) to detect more than one target nucleic acid and / or to determine the concentration, and / or relative concentrations of multiple target nucleic acids within a sample. In some embodiments, digital PCR is performed as endpoint PCR (e.g., for each of the partitions, reaction vessels, etc.). In some embodiments, digital PCR is performed as rtPCR (e.g., for each of the partitions, reaction vessels, etc.). In some embodiments, digital PCR is performed as dPCR LATE (e.g., for each of the partitions, reaction vessels, etc.). In some embodiments, digital PCR is performed as cdPCR (e.g., for each of the partitions). Conventional PCR theoretically results in an exponential amplification of a nucleic acid sequence (e.g. template or target nucleic acid) from a sample over a series of cycles in the early portion of the amplification process. By measuring the number of amplification cycles required to achieve a threshold level of amplification (as in real-time PCR), the starting concentration of nucleic acid can be calculated. However, there are many factors that affect the exponential amplification of the PCR process, such as varying amplification efficiencies, competition of low copy number targets with higher concentration targets for reagents, and interference with background contaminant nucleic acid. Digital PCR is generally less sensitive to these factors, since it does not rely on the assumption that the PCR process is exponential. In digital PCR, individual nucleic acid molecules from the initial sample are distributed amongst the partitions, and then amplified to detectable levels. The distribution of these target nucleic acid molecules across the partitions is governed by Poisson statistics. Quantitation in dPCR is possible whenever there is a sufficient number of 0 target containing partitions to apply Poisson statistics. As the number of 0 target containing partitions decreases approaching zero, the uncertainty in the quantitation value increases, ultimately becoming undefined (infinity) when there are no 0 target containing partitions. In embodiments in which multiple target nucleic acids are analyzed, digital PCR provides statistically relevant measure of the concentrations or ratios to multiple target nucleic acids. In some embodiments, the present disclosure provides dPCR methods, or elements thereof.Docket No. ABBTT-43595.601 In some embodiments, the present disclosure provides LATE PCR (U.S. Patent No. 7,632,642; herein incorporated by reference in its entirety), or elements thereof. In some embodiments LATE PCR methods are performed in a digital format (e.g. dPCR LATE). In some embodiments, LATE PCR methods are performed on a partitioned sample. In some embodiments, LATE PCR techniques are utilized for amplification of a partitioned sample (e.g. digital or non-digital sample). In some embodiments, LATE-PCR or dPCR LATE is a non-symmetric DNA amplification employing the polymerase chain reaction (PCR) process utilizing one oligonucleotide primer (the “Excess Primer”) in at least five-fold excess with respect to the other primer (the “Limiting Primer”), which itself is utilized at low concentration, up to 200 nM, so as to be exhausted in roughly sufficient PCR cycles to produce fluorescently detectable double-stranded amplicon, wherein the concentration- adjusted melting temperature of the Limiting Primer at the start of amplification, Tm[0]<L>, is not more than 5° C. below the concentration-adjusted melting temperature of the Excess Primer at the start of amplification, Tm[0]<X>, preferably at least as high and more preferably 3-10° C. higher; and wherein thermal cycling is continued for multiple cycles after exhaustion of the Limiting Primer to produce single-stranded product, namely, the extension product of the Excess Primer, sometimes referred to as the “Excess Primer Strand”. In some embodiments, the present disclosure provides competitive PCR (e.g. cdPCR), or elements thereof. Conventional competitive PCR is a method for target quantitation using end point PCR, which employs the co-amplification of an unknown and IS target sequences that share primer sequences. Since both amplifications consume the same primers, they compete for the available resources. If each reaction has the same efficiency, the relationship of the initial (pre-amplification) concentration of unknown (T0) and IS (IS0) to the final post amplification concentration after j cycles (Tj and ISj) is described by the expression: Log(Tj / ISj)=log(T0)−log(IS0) In conventional competitive PCR applications, utilization of this method requires a priori knowledge of IS0 or a calibration curve to establish the relationship between measured response and unknown concentration. The calibration curve is expressed as: log (target response / internal standard response) vs. log ([target]). This response produces a relatively straight-line curve that allows for subsequent reactions to be quantitated. The curve deviates from linearity based on how similar the amplification efficiencies are for the 2 competitive targets. In general, this method can produce up to approximately 5 logs of quantitation depending on the precision of the response measurements. The problems for quantitation associated with the conventional competitive PCR method include limited dynamic range and imprecision at the extremes of the quantitative range. In addition, this method requires generation of the calibration curve described above due to variation in IS concentration from lot to lot, reaction toDocket No. ABBTT-43595.601 reaction, and other factors. In some embodiments, these challenges are overcome by performing competitive PCR in a digital format. In some embodiments, the present disclosure utilizes real-time PCR, or elements thereof. In some embodiments, real-time PCR achieves quantitation by reading the fluorescence response of the target PCR reaction at frequent cycle intervals (e.g. every cycle), and identifying the cycle number at which the signal crosses a threshold response value. Quantitation is based on the efficiency of amplification and the number of cycles or amplification events required to produce a detectable concentration of product. The problems for quantitation associated with real-time PCR largely relate to determination of low copy targets in the presence of other amplifications, such as for other high titer targets or nonspecific amplifications, e.g., primer dimers. Other challenges include amplification in the presence of non-amplifying nucleic acid (e.g. genomic DNA) that can interfere with the amplification process and suppress low copy amplification responses. Real-time PCR also requires some type of calibration, typically either a calibration curve run within the batch or stored, or a reference quantitation standard (QS) of known concentration run in every reaction that defines the relationship between cycle number and concentration. In some embodiments, the present disclosure provides amplification techniques (e.g. dPCR LATE, cdPCR, combinations thereof, etc.) capable of extending the dynamic range for dPCR and achieving precise quantitation at and beyond the upper end of the quantitative range. In some embodiments, amplification and analysis methods described herein (e.g. dPCR LATE, cdPCR, combinations thereof, etc.) provide an extension of the dPCR dynamic range to as much as much as 8 orders of magnitude (e.g.5 orders of magnitude ...6 orders of magnitude ...7 orders of magnitude . ..8 orders of magnitude). In some embodiments, amplification and analysis methods described herein (e.g. dPCR LATE, cdPCR, combinations thereof, etc.) provide an extension of the dPCR dynamic range by up to three logs over what is obtainable by Poisson analysis (e.g., extension of 1 order of magnitude, extension of 2 orders of magnitude, extension of 3 orders of magnitude). 9. Amplicon detection / quantification In some embodiments, a sample is partitioned using any suitable method known in the art, and a nucleic acid amplification procedure (e.g. cdPCR, dPCR LATE, etc.) is performed to amplify target nucleic acids present in one or more of the partitions. The partitions can include microparticles dispersed within a volume of liquid within the one or more partitions to calibrate the volume of liquid. The one or more partitions may include probes that are specific for a single target nucleic acid of interest. In some embodiments, a sample is aliquoted into a series of tubes or reaction vessels using any suitable method known in the art, and a nucleic acid amplification procedure (e.g., cdPCR, dPCR, LATE, isothermal amplification, etc.) is performed to amplify target nucleic acids present in one or more of the tubes or reaction vessels. The tubes or reaction vessels can include microparticlesDocket No. ABBTT-43595.601 dispersed within a volume of liquid within the one or more tubes or reaction vessels to calibrate the volume of liquid. The one or more tubes or reaction vessels may include probes that are specific for a single target nucleic acid of interest. In some embodiments, a quantification method is provided to quantify the amplicons produced by the amplification reactions in each partition. In some embodiments, quantification is performed following amplification (e.g., end-point detection). In some embodiments quantification is performed in real time (e.g. amplicon quantification following each round of amplification). In some embodiments, the present disclosure provides systems, devices, methods, and compositions to detect the presence of and / or quantify nucleic acids (e.g. amplicons (e.g. IS amplicons, target amplicons), labeled nucleic acids) in a sample or partition. In some embodiments, the present disclosure provides detection of the presence of amplicons in partitions. In some embodiments, the present disclosure provides quantification of amplicons (e.g. IS amplicons, target amplicons) in a partition. In some embodiments, the present disclosure provides relative quantification of target and IS amplicons. In some embodiments, amplicon quantification involves measurement or detection of a characteristic of partitions and / or amplicons, such as a physical, chemical, luminescence, or electrical aspect, which correlates with amplification (e.g. fluorescence, luminescence, radioactivity, or other detectable signal) and allows amplicon quantification. In some embodiments, amplicon quantification is performed by a fluorescence detection technique. In some embodiments, fluorescence detection methods are provided for detection / quantification of amplified nucleic acids. In addition to the reagents already discussed, and those known to those of skill in the art of nucleic acid amplification and quantification, various detection reagents, such as fluorescent and non-fluorescent dyes and probes are provided. For example, the protocols may employ reagents suitable for use in a TaqMan™ reaction, such as a TaqMan™ probe; reagents suitable for use in a SYBR Green fluorescence detection; reagents suitable for use in a molecular beacon reaction, such as molecular beacon probes; reagents suitable for use in a scorpion reaction, such as a scorpion probe; reagents suitable for use in a fluorescent DNA-binding dye-type reaction, such as a fluorescent probe; and / or reagents for use in a LightUp protocol, such as a LightUp probe. In some embodiments, the present disclosure provides methods and compositions for quantifying a detectable signal (e.g. fluorescence) from partitions containing amplified nucleic acid (e.g. target amplicons, IS amplicons, etc.). Thus, for example, methods may employ labeling (e.g. during amplification, post-amplification) amplified nucleic acids with a detectable label, exposing partitions to a light source at a wavelength selected to cause the amplicon bound probe dye to fluoresce, and detecting and / or measuring the resulting fluorescence. Fluorescence emitted from the partitions can be tracked during amplification reaction to permit monitoring of the reaction (e.g., using a SYBR Green-type compound), or fluorescence can be measured post-amplification.Docket No. ABBTT-43595.601 In some embodiments, the present disclosure provides methods of detecting and / or quantifying the presence of a target nucleic acid in partitions by providing a probe with specificity for a target nucleic acid (e.g., a TaqMan™-type probe) in partitioned amplification reactions, and detecting / measuring the resulting fluorescence. In some embodiments, partitions containing amplified nucleic acid (e.g. target amplicons, IS amplicons, etc.) will exhibit quantifiable post-amplification fluorescence. In some embodiments, detection of a fluorescent signal is indicative of the presence of the template nucleic acid (e.g. IS, target) in the partition. The present disclosure provides corresponding methods for using other suitable target- specific probes (e.g., for target nucleic acids of interest, intercalation dyes, scorpion probes, molecular beacons, etc.), as would be understood by one of skill in the art. In some embodiments, the present disclosure provides quantification of amplified nucleic acids using one or more of fluorescent labeling, fluorescent intercalation dyes, FRET-based detection methods (U.S. Patent No.5,945,283; PCT Publication WO 97 / 22719; both of which are incorporated by reference in their entireties), quantitative PCR, real-time fluorogenic methods (U.S. Patent No.5,210,015 to Gelfand, U.S. Patent No.5,538,848 to Livak, et al., and U.S. Patent No.5,863,736 to Haaland, as well as Heid, C. A., et al., Genome Research, 6:986-994 (1996); Gibson, U. E. M, et al., Genome Research 6:995-1001 (1996); Holland, P. M., et al., Proc. Natl. Acad. Sci. USA 88:7276-7280, (1991); and Livak, K. J., et al., PCR Methods and Applications 357-362 (1995), each of which is incorporated by reference in its entirety), molecular beacons (Piatek, A. S., et al., Nat. Biotechnol.16:359-63 (1998); Tyagi, S. and Kramer, F. R., Nature Biotechnology 14:303-308 (1996); and Tyagi, S. et al., Nat. Biotechnol.16:49-53 (1998); herein incorporated by reference in their entireties), Invader™ assays (Third Wave Technologies, (Madison, Wis.)) (Neri, B. P., et al., Advances in Nucleic Acid and Protein Analysis 3826:117-125, 2000; herein incorporated by reference in its entirety), nucleic acid sequence-based amplification (NASBA; (See, e.g., Compton, J. Nucleic Acid Sequence-based Amplification, Nature 350: 91-91, 1991.; herein incorporated by reference in its entirety), Scorpion probes (Thelwell, et al. Nucleic Acids Research, 28:3752-3761, 2000; herein incorporated by reference in its entirety), capacitive DNA detection (See, e.g., Sohn, et al. (2000) Proc. Natl. Acad. Sci. U.S.A.97:10687-10690; herein incorporated by reference in its entirety), etc. In some embodiments, labeling methods are used to differentially label different amplicons. In some embodiments, IS amplicons and target amplicons are differentially labeled to allow for separate detection and quantification. In some embodiments, IS amplicons and target amplicons are differentially labeled to allow for sorting of IS-containing partitions from partitions without IS amplicons. In some embodiments, different target sequences are differentially labeled to allow for separate detection and quantification. In some embodiments, different target sequences are differentially labeled to allow for sorting of one target from another. In some embodiments,Docket No. ABBTT-43595.601 differential labeling is achieved via sequence specific labeling probes with detectably different fluorescent labels. 10. High-throughput and automated screening The present disclosure contemplates the use of the volume calibrators (e.g., predetermined number of microparticles) in high-throughput and automated screening processes. It is desirable in a variety of applications to measure and analyze the behavior of a large number of tissue and / or cell samples (e.g., dozens, hundreds, or thousands of samples representing variation across a variety of experimental conditions). Such analysis can facilitate the assessment of a variety of therapeutic treatments, environmental conditions, genetic modifications, communicable diseases, or other specified conditions to assess the efficacy of a treatment, to determine the toxicity of a substance, or to determine some other information of interest. For example, such high-throughput screening of hundreds or thousands of different samples can facilitate the identification of high-growth clone cell lines, while reducing the cost and / or time associated with such processes. This detection and analysis can include extracting small volumes from each sample to perform flow cytometry and generating cell counts or other information about the population(s) of cells in each sample, such as cell health and consistency of the sample. A robotic sample handling apparatus can be used to reduce the cost and / or time required to perform such assessments on many samples. Such “high throughput screening” apparatus and methods can include using multi-well sample plates or other multi-sample containers that permit robots or other apparatus to interact with dozens, hundreds, or thousands of samples in a single container in an automated manner. This can include placing a multi-well sample plate into an automated flow cytometry instrument that includes a robotic sample extraction head. Such a sample extraction head can move in two or more dimensions, facilitating the extraction of fluid from samples across the multi-well sample plate, as well as extracting chemical reagents, flushing of cleaning fluids, or other substances from reservoirs located nearby. The high throughput screening, volumetric flow cytometry systems, are examples of such a robotic sample handling apparatus that can provide fast, low-cost screening of samples. An additional robotic arm or other automated sample handling apparatus can also be used to present multiple different multi-well sample plates to the flow cytometry instrument over time, facilitating the automated assessment of even more samples with no or minimal human intervention. Such a system can operate to screen many samples over an extended period of time without human intervention, e.g., for longer than 48 hours. Such a combined system can include expanded reservoirs for rinse fluids, cleaning fluids, or other consumables. Additionally or alternatively, local reservoirs that are accessible by a sample head of a robotic sample handling apparatus can be augmented by additional remote reservoirs that are operable to automatically replenish the local reservoirs.Docket No. ABBTT-43595.601 In a high-throughput and automated screening system, a predetermined number of microparticles can be dispersed in one or more wells of a multi-well sample plate to calibrate the volume of liquid in the one or more wells of the sample plate to enable accurate volume measurements of aliquots taken from the one or more wells. For example, the volume measurements can be detected using a flow cytometry (e.g., volumetric) to count the beads in the aliquoted sample and a computer can be used to determine the volume of the aliquots using Equation 1. 11. Methods for measuring the level of analyte of interest or biomarker In the methods described above, an analyte of interest or biomarker levels can be measured by any means, such as antibody dependent methods, such as immunoassays, protein immunoprecipitation, immunoelectrophoresis, chemical analysis, SDS-PAGE and Western blot analysis, or protein immunostaining, electrophoresis analysis, a protein assay, a competitive binding assay, a functional protein assay, or chromatography or spectrometry methods, such as high- performance liquid chromatography (HPLC) or liquid chromatography–mass spectrometry (LC / MS). Also, the assay can be employed in clinical chemistry format such as would be known by one skilled in the art. In some embodiments, the sample is diluted or undiluted. The sample can be from about 1 to about 25 microliters, about 1 to about 24 microliters, about 1 to about 23 microliters, about 1 to about 22 microliters, about 1 to about 21 microliters, about 1 to about 20 microliters, about 1 to about 18 microliters, about 1 to about 17 microliters, about 1 to about 16 microliters, about 15 microliters or about 1 microliter, about 2 microliters, about 3 microliters, about 4 microliters, about 5 microliters, about 6 microliters, about 7 microliters, about 8 microliters, about 9 microliters, about 10 microliters, about 11 microliters, about 12 microliters, about 13 microliters, about 14 microliters, about 15 microliters, about 16 microliters, about 17 microliters, about 18 microliters, about 19 microliters, about 20 microliters, about 21 microliters, about 22 microliters, about 23 microliters, about 24 microliters or about 25 microliters. In some embodiments, the sample is from about 1 to about 150 microliters or less or from about 1 to about 25 microliters or less. Other methods of detection include the use of or can be adapted for use on a nanopore device or nanowell device. Examples of nanopore devices are described in International Patent Publication No. WO 2016 / 161402, which is hereby incorporated by reference in its entirety. Examples of nanowell device are described in International Patent Publication No. WO 2016 / 161400, which is hereby incorporated by reference in its entirety. 12. Variations on methods The methods may also be adapted in view of other methods for analyzing analytes. Examples of well-known variations include, but are not limited to, immunoassay, such as sandwichDocket No. ABBTT-43595.601 immunoassay (e.g., monoclonal-monoclonal sandwich immunoassays, monoclonal-polyclonal sandwich immunoassays, including enzyme detection (enzyme immunoassay (EIA) or enzyme-linked immunosorbent assay (ELISA), competitive inhibition immunoassay (e.g., forward and reverse), enzyme multiplied immunoassay technique (EMIT), a competitive binding assay, bioluminescence resonance energy transfer (BRET), one-step antibody detection assay, homogeneous assay, heterogeneous assay, capture on the fly assay, etc. The presence or amount of analyte (e.g., biomarker) present in a body sample may be readily determined using an immunoassay, such as sandwich immunoassay (e.g., monoclonal- monoclonal sandwich immunoassays, monoclonal-polyclonal sandwich immunoassays, including radioisotope detection (radioimmunoassay (RIA)) and enzyme detection (enzyme immunoassay (EIA) or enzyme-linked immunosorbent assay (ELISA) (e.g., Quantikine ELISA assays, R&D Systems, Minneapolis, MN)). An example of a point-of-care device that can be used is i-STAT® (Abbott, Laboratories, Abbott Park, IL). Other methods that can be used include a chemiluminescent microparticle immunoassay, in particular one employing the ARCHITECT® automated analyzer (Abbott Laboratories, Abbott Park, IL), as an example. Other methods include, for example, mass spectrometry, and immunohistochemistry (e.g., with sections from tissue biopsies), using anti-analyte antibodies (monoclonal, polyclonal, chimeric, humanized, human, etc.) or antibody fragments thereof against analyte. Other methods of detection include those described in, for example, U.S. Patent Nos. 6,143,576; 6,113,855; 6,019,944; 5,985,579; 5,947,124; 5,939,272; 5,922,615; 5,885,527; 5,851,776; 5,824,799; 5,679,526; 5,525,524; and 5,480,792, each of which is hereby incorporated by reference in its entirety. Specific immunological binding of the antibody to the analyte can be detected via direct labels, such as fluorescent or luminescent tags, metals and radionuclides attached to the antibody or via indirect labels, such as alkaline phosphatase or horseradish peroxidase. The use of immobilized antibodies or antibody fragments thereof may be incorporated into the immunoassay. The antibodies may be immobilized onto a variety of supports, such as magnetic or chromatographic matrix particles, the surface of an assay plate (such as microtiter wells), pieces of a solid substrate material, and the like. An assay strip can be prepared by coating the antibody or plurality of antibodies in an array on a solid support. Any solid support known in the art can be used, including but not limited to, solid supports made from polymeric materials in the forms of wells, tubes or beads. This strip can then be dipped into the test sample and processed quickly through washes and detection steps to generate a measurable signal, such as a colored spot. A homogeneous format may be used. For example, after the test sample is obtained from a subject, a mixture is prepared. The mixture contains the test sample being assessed for analyte (e.g., biomarker), a first specific binding partner, and a second specific binding partner. The order in which the test sample, the first specific binding partner, and the second specific binding partner are added to form the mixture is not critical. The test sample is simultaneously contacted with the first specificDocket No. ABBTT-43595.601 binding partner and the second specific binding partner. In some embodiments, the first specific binding partner and any biomarker contained in the test sample may form a first specific binding partner-analyte-antigen complex and the second specific binding partner may form a first specific binding partner-analyte of interest (e.g., biomarker)-second specific binding partner complex. The first specific binding partner may be an anti-analyte antibody. The second specific binding partner may be an anti-analyte antibody. Moreover, the second specific binding partner is labeled with or contains a detectable label as described above. A heterogeneous format may be used. For example, after the test sample is obtained from a subject, a first mixture is prepared. The mixture contains the test sample being assessed for analyte (e.g., biomarker) and a first specific binding partner, wherein the first specific binding partner and any biomarker contained in the test sample form a first specific binding partner-analyte-antigen complex. The order in which the test sample and the first specific binding partner are added to form the mixture is not critical. In those embodiments where the solid phase is a bead, the bead may be a magnetic bead or a magnetic particle. Magnetic beads / particles may be ferromagnetic, ferrimagnetic, paramagnetic, superparamagnetic or ferrofluidic. Exemplary ferromagnetic materials include Fe, Co, Ni, Gd, Dy, CrO2, MnAs, MnBi, EuO, and NiO / Fe. Examples of ferrimagnetic materials include NiFe2O4, CoFe2O4, Fe3O4 (or FeO.Fe2O3). Beads can have a solid core portion that is magnetic and is surrounded by one or more non-magnetic layers. Alternately, the magnetic portion can be a layer around a non-magnetic core. The beads on which the first specific binding member is immobilized may be stored in dry form or in a liquid. The magnetic beads may be subjected to a magnetic field prior to or after contacting with the sample with a magnetic bead on which the first specific binding member is immobilized. 13. Kits The systems and methods herein can be implemented using a kit of the disclosure. In an aspect, a kit is provided for calibrating a volume of a liquid in a sample to determine the volume of an aliquot of the sample. The kit may include a predetermined number of microparticles, a counting device to count an indeterminate number of microparticles in an aliquot of the sample, and instructions for calibrating a predetermined volume of a liquid in a sample to determine the volume of an aliquot of the sample. The instructions inform a user of the kit to disperse the predetermined number of microparticles into a predetermined volume of liquid in a sample to calibrate the volume of liquid in the sample based on the number of microparticles per µL of liquid in the sample; and calculate the volume of liquid in an aliquot of the sample by multiplying the predetermined volume of liquid in the sample by the quotient of a counted number of the indeterminate number of microparticles in the aliquot of the sample divided by the predetermined number of microparticles dispersed in the sample.Docket No. ABBTT-43595.601 The kit can also include a sample collection device (e.g., a microsampling device, a syringe, etc.) and instructions for collecting a sample using the collection device. The kit can also include a counting instrument and instructions for counting the indeterminate number of microparticles in the aliquot using the instrument. The kit can also include instruments for performing amplification assays and amplification reagents. In some instances, the kit includes a reaction vessel or plurality of reaction vessels. The kit can also provide instructions for amplifying in a reaction vessel a target nucleic acid. The kit can also include instructions for detecting in a reaction vessel the target nucleic acid. The kit can also include instruments or equipment for performing a digital immunoassay. The kit can also include instructions for performing a digital immunoassay for a target antigen. The kit can also include instructions for calculating the dilution factor of the volume of liquid. Provided herein is a kit, which may be used for assaying or assessing a test sample for an analyte of interest. The kit comprises at least one component for assaying the test sample for the analyte of interest, and instructions for assaying the test sample for the analyte of interest. For example, the kit can comprise instructions for assaying the test sample for an analyte of interest by immunoassay, e.g., chemiluminescent microparticle immunoassay. Instructions included in kits can be affixed to packaging material or can be included as a package insert. While the instructions are typically written or printed materials, they are not limited to such. Any medium capable of storing such instructions and communicating them to an end user is contemplated by this disclosure. Such media include, but are not limited to, electronic storage media (e.g., magnetic discs, tapes, cartridges, chips), optical media (e.g., CD ROM), and the like. As used herein, the term "instructions" can include the address of an internet site that provides the instructions. Alternatively or additionally, the kit can comprise a calibrator or control, e.g., purified, and optionally lyophilized, and / or at least one container (e.g., tube, microtiter plates or strips) for conducting the assay, and / or a buffer, such as an assay buffer or a wash buffer, either one of which can be provided as a concentrated solution, a substrate solution for the detectable label, or a stop solution. Preferably, the kit comprises all components, i.e., reagents, standards, buffers, diluents, etc., which are necessary to perform the assay. The instructions also can include instructions for generating a standard curve. Moreover, the kit can further comprise a microsampling device, a plasma separation device and / or a microsampling device and a plasma separation device. Optionally, the kit includes quality control components (for example, sensitivity panels, calibrators, and positive controls). Preparation of quality control reagents is well-known in the art and is described on insert sheets for a variety of immunodiagnostic products. Sensitivity panel membersDocket No. ABBTT-43595.601 optionally are used to establish assay performance characteristics, and further optionally are useful indicators of the integrity of the immunoassay kit reagents, and the standardization of assays, The kit can also optionally include other reagents required to conduct a diagnostic assay or facilitate quality control evaluations, such as buffers, salts, enzymes, enzyme co-factors, substrates, detection reagents, and the like. Other components, such as buffers and solutions for the isolation and / or treatment of a test sample (e.g., pretreatment reagents), also can be included in the kit. The kit can additionally include one or more other controls. One or more of the components of the kit can be lyophilized, in which case the kit can further comprise reagents suitable for the reconstitution of the lyophilized components. The various components of the kit optionally are provided in suitable containers as necessary, e.g., a microtiter plate. The kit can further include containers for holding or storing a sample (e.g., a container or cartridge for a urine, whole blood, plasma, or serum sample). Where appropriate, the kit optionally also can contain reaction vessels, mixing vessels, and other components that facilitate the preparation of reagents or the test sample. The kit can also include one or more instrument for assisting with obtaining a test sample, such as a syringe, pipette, forceps, measured spoon, or the like. If the detectable label is at least one acridinium compound, the kit can comprise at least one acridinium-9-carboxamide, at least one acridinium-9-carboxylate aryl ester, or any combination thereof. If the detectable label is at least one acridinium compound, the kit also can comprise a source of hydrogen peroxide, such as a buffer, solution, and / or at least one basic solution. If desired, the kit can contain a solid phase, such as a magnetic particle, bead, test tube, microtiter plate, cuvette, membrane, scaffolding molecule, film, filter paper, disc, or chip. If desired, the kit can further comprise one or more components, alone or in further combination with instructions, for assaying the test sample for another analyte, which can be a biomarker, such as a biomarker of traumatic brain injury or disorder. It will be readily apparent to those skilled in the art that other suitable modifications and adaptations of the methods of the present disclosure described herein are readily applicable and appreciable, and may be made using suitable equivalents without departing from the scope of the present disclosure or the aspects and embodiments disclosed herein. Having now described the present disclosure in detail, the same will be more clearly understood by reference to the following non-limiting example(s), which are merely intended only to illustrate some aspects and embodiments of the disclosure, and should not be viewed as limiting to the scope of the disclosure. The disclosures of all journal references, U.S. patents, and publications referred to herein are hereby incorporated by reference in their entireties.Docket No. ABBTT-43595.601 EXAMPLES Example 1

[0004] Accurately and precisely aliquoting small volumes of clinical samples (<10 µL) is challenging due to the imprecise dimensions of devices such as manual or automated pipettes, microfluidic flow chambers, or other volumetric tools. Instead of controlling the flow of the sample, the inherent error in liquid delivery can be compensated for by independently determining the delivered volume using volume calibrators. FIG. 1 details how to calculate an unknown aliquot volume using labeled microparticles or beads. As shown in FIG.1, a homogeneous liquid suspension (e.g., water or buffer) containing a known number of labeled microparticles or beads is prepared (“Volume Calibrator Beads”). The volume of liquid containing the Volume Calibrator Beads is known. The microparticles or beads can be labeled with a fluorescent dye and / or with a radionuclide (e.g., such as Co-57, Co-60, Cs-137, Eu-152, Ga-67, I-125, I-132, In-111, Mo-99, P-32, Sm-153, Tc-99m, or Tl-201). If a radionuclide is used, the microparticles or beads can be counted with a Geiger counter.

[0005] A known quantity (e.g., volume) of a first sample, such as a biological sample, is provided. As shown in FIG. 1, the known volume of the first sample is 1000 µL. To the first sample, a small volume (the amount is not critical – it can be insignificant from a mathematical perspective) of the liquid containing the Volume Calibrator Beads is added to the first sample. The number of Volume Calibrator Beads added to the first sample from the liquid containing the Volume Calibrator Beads is known. As shown in FIG.1, the number of Volume Calibrator Beads in the small volume of the liquid containing the Volume Calibrator Beads is 10. The step of adding a small volume of liquid containing a known number of Volume Calibrator Beads to the first sample calibrates the first volume of liquid in the first sample based on the number of microparticles per µL of liquid in this sample (“Calibrated First Sample”). An unknown or indeterminate aliquot volume of the Calibrated First Sample is taken and used to create a second sample. The volume of the aliquoted sample in the second sample is determined using the Equation 1 below:As shown in FIG.1, if 5 beads are counted in the second sample, the aliquot volume is determined by: 5 beads in the second sample / 10 beads in the Calibrated First Sample x 1000 µL (the volume of the first sample) to provide an aliquot volume of 500 µL. As shown in Example 2, if 3 beads are counted in the second sample, the aliquot volume is determined by: 3 beads in the second sample / 10 beads in the Calibrated First Sample x 1000 µL (the volume of the first sample) to provide an aliquot volume of 300 µL. As shown in Example 3, if 1 bead is counted in the second sample, the aliquot volume is determined by: 1 bead in the second sample / 10 beads in the Calibrated First Sample” x 1000 µL (the volume of the first sample) to provide an aliquot volume of 100 µL.Docket No. ABBTT-43595.601 In some aspects, using the volume calibrator in liquid form can be more effective when its volume is significantly less than that of the original sample (e.g., less than 10-fold to 1,000 fold less). In other aspects, the calibrator can also be added dry to the original sample. It is understood that the foregoing detailed description and accompanying examples are merely illustrative and are not to be taken as limitations upon the scope of the disclosure, which is defined solely by the appended claims and their equivalents. Various changes and modifications to the disclosed embodiments will be apparent to those skilled in the art. Such changes and modifications, including without limitation those relating to the chemical structures, substituents, derivatives, intermediates, syntheses, compositions, formulations, or methods of use of the disclosure, may be made without departing from the spirit and scope thereof. For reasons of completeness, various aspects of the disclosure are set out in the following numbered clauses: Clause 1. A method of determining a volume of a liquid in a sample, comprising: (a) dispersing a predetermined number of microparticles per unit of a predetermined first volume of liquid into a first sample; (b) providing a second sample comprising an indeterminate second volume of liquid obtained from the predetermined first volume of liquid; and (c) determining the indeterminate second volume of liquid in the second sample based on the number of microparticles in the second sample. Clause 2. The method of clause 1, further comprising counting the number of microparticles in the first and / or second sample. Clause 3. The method of clause 2, wherein the indeterminate second volume of liquid is determined by an equation, wherein the equation is: Volume = (number of microparticles counted in the second sample / predetermined number of microparticles dispersed in the first sample) x (predetermined first volume of the liquid in the first sample). Clause 4. The method of any one of clauses 1 to 3, wherein the microparticles comprise inert microparticles. Clause 5. The method of any one of clauses 1 to 4, wherein the microparticles comprise monodisperse microparticles. Clause 6. The method of any one of clauses 1 to 5, wherein the microparticles comprise beads. Clause 7. The method of any one of clauses 1 to 6, wherein the microparticles comprise colored or fluorescent beads. Clause 8. The method of any one of clauses 1 to 7, wherein the microparticles are counted using a flow cytometer, a fluorescence-based counter, an optical absorbed counter, a scattering counter, refractive index counter, a resistance-based counter, a capacitance-based counter, an inductance-based counter, an impedance-based counters, a nanopore sensor, a photoacoustic-basedDocket No. ABBTT-43595.601 counter, a magnetic based counter, microscopy, particle image velocimetry, a thermal-based counter, or a vision-based automated counter. Clause 9. The method of any one of clauses 1 to 8, wherein the microparticles comprise a uniform diameter of less than about 0.10 µm, less than about 0.20 µm, less than about 0.30 µm, less than about 0.40 µm, less than about 0.50 µm, less than about 0.60 µm, less than about 0.70 µm, less than about 0.80 µm, less than about 0.9 µm, less than about 1 µm, less than about 2 µm, less than about 3 µm, less than about 4 µm, or less than about 5 µm. Clause 10. The method of any one of clauses 1 to 9, wherein the microparticles comprise a radionuclide. Clause 11. The method of clause 10, wherein the radionuclides are counted using a Geiger counter. Clause 12. The method of clauses 10 or 11, wherein the radionuclide is selected from the group consisting of Co-57, Co-60, Cs-137, Eu-152, Ga-67, I-125, I-132, In-111, Mo-99, P-32, Sm- 153, Tc-99m, Tl-201.. Clause 13. The method of any one of clauses 1 to 12, wherein predetermined number of microparticles per unit of the volume of liquid is between 1 and 10000, 100 and 5000, 500 and 2000, 750 and 1500, 800 and 1400, 900 and 1100, or between 950 and 1050 microparticles per unit of the volume of liquid. Clause 14. The method of any one of clauses 1 to 13, wherein the predetermined number of microparticles per unit of the volume of liquid is about 1000 microparticles per µL of liquid. Clause 15. The method of any one of clauses 1 to 14, wherein the first sample comprises a bronchial lavage sample, a cerebrospinal fluid sample, a crevicular sample, an interstitial fluid sample, a saliva sample, a whole blood sample, or a urine sample. Clause 16. The method of any one of clauses 1 to 15, wherein the first sample comprises a suspension. Clause 17. The method of any one of clauses 1 to 16, wherein the predetermined first volume of liquid is between about 1µL and about 10 mL. Clause 18. The method of any one of clauses 1 to 17, wherein the indeterminate second volume of liquid is between about 10 nL and about 100 µL. Clause 19. The method of any one of clauses 1 to 18, wherein the second sample is provided by aliquoting the indeterminate second volume of liquid from the first sample into a reaction vessel. Clause 20. A method of calibrating a volume of a liquid in a sample to determine the volume of an aliquot of the sample, comprising: (a) dispersing a predetermined number of microparticles per unit volume of a predetermined first volume of liquid in a first sample, thereby calibrating the first volume of liquid in the first sample based on the number of microparticles per µLDocket No. ABBTT-43595.601 of liquid in the first sample; (b) aliquoting an indeterminate second volume of liquid from the first sample into a container to provide a second sample comprising a number of microparticles per µL of liquid that is proportionate to the number of microparticles per µL of liquid in the first sample; and (c) determining the volume of liquid in the second sample based on the number of microparticles in the second sample, wherein the volume of liquid in the second sample is equal to the volume obtained by multiplying the predetermined first volume of liquid in the first sample by the quotient of the number of microparticles in the second sample divided by the predetermined number of microparticles dispersed in the first sample. Clause 21. A kit for calibrating a volume of a liquid in a sample to determine the volume of an aliquot of the sample, comprising: (a) a predetermined number of microparticles; and (b) a counting device to count an indeterminate number of microparticles in an aliquot of the sample; and (b) instructions for calibrating a predetermined volume of a liquid in a sample to determine the volume of an aliquot of the sample, wherein the instructions instruct a user of the kit to: (i) disperse the predetermined number of microparticles into a predetermined volume of liquid in a sample to calibrate the volume of liquid in the sample based on the number of microparticles per µL of liquid in the sample; and (ii) calculate the volume of liquid in an aliquot of the sample by multiplying the predetermined volume of liquid in the sample by the quotient of a counted number of the indeterminate number of microparticles in the aliquot of the sample divided by the predetermined number of microparticles dispersed in the sample. Clause 22. The kit of clause 21, further comprising a sample collection device. Clause 23. The kit of clause 22, further comprising instructions for collecting a sample using the collection device. Clause 24. The kit of clause 21, further comprising instructions for counting the indeterminate number of microparticles in the aliquot using the device.

Claims

Docket No. ABBTT-43595.601 CLAIMS What is claimed is:

1. A method of determining a volume of a liquid in a sample, comprising: (a) dispersing a predetermined number of microparticles per unit of a predetermined first volume of liquid into a first sample; (b) providing a second sample comprising an indeterminate second volume of liquid obtained from the predetermined first volume of liquid; and (c) determining the indeterminate second volume of liquid in the second sample based on the number of microparticles in the second sample.

2. The method of claim 1, further comprising counting the number of microparticles in the first and / or second sample.

3. The method of claim 2, wherein the indeterminate second volume of liquid is determined by an equation, wherein the equation is: Volume = (number of microparticles counted in the second sample / predetermined number of microparticles dispersed in the first sample) x (predetermined first volume of the liquid in the first sample) 4. The method of any one of claims 1 to 3, wherein the microparticles comprise inert microparticles.

5. The method of any one of claims 1 to 4, wherein the microparticles comprise monodisperse microparticles.

6. The method of any one of claims 1 to 5, wherein the microparticles comprise beads.

7. The method of any one of claims 1 to 6, wherein the microparticles comprise colored or fluorescent beads.

8. The method of any one of claims 1 to 7, wherein the microparticles are counted using a flow cytometer, a fluorescence-based counter, an optical absorbed counter, a scattering counter, refractive index counter, a resistance-based counter, a capacitance-based counter, an inductance-based counter, an impedance-based counters, a nanopore sensor, a photoacoustic-based counter, a magnetic based counter, microscopy, particle image velocimetry, a thermal-based counter, or a vision-based automated counter.Docket No. ABBTT-43595.601 9. The method of any one of claims 1 to 8, wherein the microparticles comprise a uniform diameter of less than about 0.10 µm, less than about 0.20 µm, less than about 0.30 µm, less than about 0.40 µm, less than about 0.50 µm, less than about 0.60 µm, less than about 0.70 µm, less than about 0.80 µm, less than about 0.9 µm, less than about 1 µm, less than about 2 µm, less than about 3 µm, less than about 4 µm, or less than about 5 µm.

10. The method of any one of claims 1 to 9, wherein the microparticles comprise a radionuclide.

11. The method of claim 10, wherein the radionuclides are counted using a Geiger counter.

12. The method of claims 10 or 11, wherein the radionuclide is selected from the group consisting of Co-57, Co-60, Cs-137, Eu-152, Ga-67, I-125, I-132, In-111, Mo-99, P-32, Sm-153, Tc-99m, Tl- 201.

13. The method of any one of claims 1 to 12, wherein predetermined number of microparticles per unit of the volume of liquid is between 1 and 10000, 100 and 5000, 500 and 2000, 750 and 1500, 800 and 1400, 900 and 1100, or between 950 and 1050 microparticles per unit of the volume of liquid.

14. The method of any one of claims 1 to 13, wherein the predetermined number of microparticles per unit of the volume of liquid is about 1000 microparticles per µL of liquid.

15. The method of any one of claims 1 to 14, wherein the first sample comprises a bronchial lavage sample, a cerebrospinal fluid sample, a crevicular sample, an interstitial fluid sample, a saliva sample, a whole blood sample, or a urine sample.

16. The method of any one of claims 1 to 15, wherein the first sample comprises a suspension.

17. The method of any one of claims 1 to 16, wherein the predetermined first volume of liquid is between about 1µL and about 10 mL.

18. The method of any one of claims 1 to 17, wherein the indeterminate second volume of liquid is between about 10 nL and about 100 µL.

19. The method of any one of claims 1 to 18, wherein the second sample is provided by aliquoting the indeterminate second volume of liquid from the first sample into a reaction vessel.

20. A method of calibrating a volume of a liquid in a sample to determine the volume of an aliquot of the sample, comprising: (a) dispersing a predetermined number of microparticles per unit volume of a predetermined first volume of liquid in a first sample, thereby calibrating the first volume of liquid in the first sample based on the number of microparticles per µL of liquid in the first sample; (b) aliquoting an indeterminate second volume of liquid from the first sample into a container to provide a second sample comprising a number of microparticles per µL of liquid that is proportionate to the number of microparticles per µL of liquid in the first sample; andDocket No. ABBTT-43595.601 (c) determining the volume of liquid in the second sample based on the number of microparticles in the second sample, wherein the volume of liquid in the second sample is equal to the volume obtained by multiplying the predetermined first volume of liquid in the first sample by the quotient of the number of microparticles in the second sample divided by the predetermined number of microparticles dispersed in the first sample.

21. A kit for calibrating a volume of a liquid in a sample to determine the volume of an aliquot of the sample, comprising: (a) a predetermined number of microparticles; and (b) a counting device to count an indeterminate number of microparticles in an aliquot of the sample; and (c) instructions for calibrating a predetermined volume of a liquid in a sample to determine the volume of an aliquot of the sample, wherein the instructions instruct a user of the kit to: (i) disperse the predetermined number of microparticles into a predetermined volume of liquid in a sample to calibrate the volume of liquid in the sample based on the number of microparticles per µL of liquid in the sample; and (ii) calculate the volume of liquid in an aliquot of the sample by multiplying the predetermined volume of liquid in the sample by the quotient of a counted number of the indeterminate number of microparticles in the aliquot of the sample divided by the predetermined number of microparticles dispersed in the sample.

22. The kit of claim 21, further comprising a sample collection device.

23. The kit of claim 22, further comprising instructions for collecting a sample using the collection device.

24. The kit of claim 21, further comprising instructions for counting the indeterminate number of microparticles in the aliquot using the device.

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