Qc particles and methods of use

Abstract

The present disclosure provides systems, methods, and kits to improve the characterization, including the quality control, of a flow cytometer. In an embodiment, a system for characterization of a flow cytometer comprises a first standard particle reagent, wherein the first standard particle reagent comprises a first particle mixture and a second standard particle reagent, wherein the second standard particle reagent comprises a second particle mixture wherein the particles have a fluorescent dye.

Description

QC Particles and Usage Methods This disclosure provides systems, methods, and kits for improving the characterization, including quality control, of flow cytometry. In one embodiment, the system for characterizing a flow cytometer comprises: a first standard particle reagent, wherein the first standard particle reagent comprises a first particle mixture; and a second standard particle reagent, wherein the second standard particle reagent comprises a second particle mixture, wherein the particles have a fluorescent dye. Abstract

Claims

CLAIMSWhat is claimed is:

1. A system for characterization of a flow cytometer comprising: a first standard particle reagent, wherein the first standard particle reagent comprises a first particle mixture; and a second standard particle reagent, wherein the second standard particle reagent comprises a second particle mixture wherein the particles have a fluorescent dye.

2. The system of claim 1. wherein the first and second standard particle mixtures comprise beads or microparticles and the first and second standard particle mixture comprises a plurality7of subpopulations of particles, each subpopulation having a different average diameter.

3. The system of any one of claims 1-2, wherein the standard particle mixtures comprise a synthetic material, a metal material, a hollow sphere, a latex bead, a gold nanoparticle, a lipid nanoparticle (LNP), a polysty rene bead, a hydrogel particle, a silica particle, a Poly(methyl methacrylate) (PMMA) particle, or a combination thereof.

4. The system of any one of claims 1-3, wherein the standard particle mixtures have particles with a diameter betw een 100 nm and 1000 nm.

5. The system of any one of claims 1-4, wherein the standard particle mixtures are excited by a laser channel between 325 to 808 nm.

6. The system of any one of claims 1-5, wherein the standard particle mixtures comprise beads where at least one fluorosphere with no dye and at least one bead with a fluorescent dye.

7. The system of any one of claims 1-6, wherein the standard particle mixtures comprise beads with at least 8 peaks of fluorescent intensity.

8. The system of any one of claims 1-7, wherein the second standard particles are coated with dyes capable of being excited by lasers having the following wavelengths: 355 nm (UV), 405 nm (Violet), 488 nm (Blue), 561 nm (Y ellow-Green), 638 nm (Red), and 808 nm (Infrared).

9. A method for performing flow cytometer quality control comprising:(a) loading the first standard particle reagent and the second standard particle reagent of any one of claims 1-8 into a flow7cytometer;(b) evaluating at least one of the follow ing:(i) evaluating side scatter resolution of at least one side scatter channel of at least one laser in the flow cytometer based on a single peak analysis;(ii) evaluating gain of at least one side scatter channel of at least one laser in the flow7cytometer based on a single peak analysis;(iii) evaluating rCV of at least one laser in the flow cytometer based on a single peak analysis(iv) evaluating fluorescence sensitivity of at least one fluorescent channel of at least one laser in the flow7cytometer based on at least tw o peak fluorescence intensity analysis;(v) evaluating flow rate in the flow cytometer based on a single peak analysis; and(vi) comparing performance (day-to-day or across time of the same flow cytometer using the same first standard particle reagent and second standard particle reagent); and(c) based on the evaluations in step (b), determining at least one of the following: (i) whether the flow cytometer passes or fails quality control; (ii) details regarding current instrument component performance status.

10. The method of claim 9, wherein the evaluation in step (b) is performed with a plurality of lasers having the following wavelengths: 355 nm (UV), 405 nm (Violet), 488 nm (Blue), 561 nm (Y ellow-Green), 638 nm (Red), and 808 nm (Infrared).

11. The method of any one of claims 9-10, further comprising: generating a quality control report following step (c).

12. The method of any one of claims 9-11, further comprising the step of (d) performing an enhanced quality control analysis based on 1, 2, 4, 8, or 12 peaks of fluorescent intensity.

13. The method of any one of claims 9-12, wherein the bead mixture of claims 1-8 is run to determine a distance between noise and a highest number of peaks that can be resolved by each fluorescence channel.

14. The method of any one of claims 9-13, further comprising the step of (e) generating a quality control report to determine the sensitivity of detecting cellular structures between 30- 2000 nm.

15. The method of claim 14, further comprising, (f) loading a test a sample to detect a fluorescently labeled cellular structure with a diameter between 30 - 2000 nm.

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