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148 results about "96 well plate" patented technology

High performance liquid chromatography tandem mass spectrometry detection method of 25-hydroxy vitamin D in serum

The invention discloses a high performance liquid chromatography tandem mass spectrometry detection method of 25-hydroxy vitamin D in serum. The method comprises the following steps that (1) a sampleis pre-treated, specifically, an internal standard working solution and a sodium hydroxide solution are sequentially added into a serum sample and mixed to be uniform; (2) solid-liquid extraction treatment is conducted, specifically, the sample treated in the step (1) is loaded to an SLE 96 pore plate, and eluted with normal hexane, and then the eluant is blown to be dried by using nitrogen; (3) aderivatization reaction is conducted, a derivatization reagent 4-phenyl-1,2,4-triazoline-3,5-diketone (PTAD) solution is added to redissolve, a thermostatic reaction is conducted, then ethyl alcoholis added to terminate the reaction, and the product is blown to be dried by using nitrogen; and (4) redissolution is conducted, and analytical detection is conducted by adopting the high performance liquid chromatography tandem mass spectrometry based on a multiplexing system (MPX). The high performance liquid chromatography tandem mass spectrometry detection method is high in detection sensitivity, high in specificity and low in cost, meanwhile, the pre-treatment time and analysis time of the sample are effectively shortened, and the detection efficiency of the sample is improved.
Owner:杭州凯莱谱精准医疗检测技术有限公司

Micro-scale cell electro-transfection micro-fluidic chip, electro-transfection sorter and applications thereof

The invention relates to a micro-scale cell electro-transfection micro-fluidic chip, an electro-transfection sorter and applications thereof. The micro-scale cell electro-transfection sorter comprisesan electro-transfection unit, a display screen, an exterior box, a power supply unit, a micro control unit, and a main sensor. The electro-transfection unit comprises the chip. The display screen isused to send a command to the micro control unit and receive and display feedback information from the micro control unit and the main sensor. The micro control unit is used to receive the command sent by the display screen and control the electro-transfection unit and the power supply unit. The electro-transfection unit is used to complete cell transfection. The main sensor is used to receive feedback information from the electro-transfection unit and send the feedback information to the display screen and the micro control unit. The micro-scale cell electro-transfection micro-fluidic chip comprises an injection port, a discharge port, a negative pressure channel, a positive pressure channel, and a main channel, and a 96-well plate is arranged behind the discharge port. The provided chipcan guarantee that the transfection states in the main channel are identical during the transfection process, the transfection efficiency is guaranteed, the cell quality is guaranteed by the 96-well plate, and the subsequent cell culture becomes more convenient.
Owner:ALLIFE MEDICAL SCI & TECH CO LTD

High through-put detection of pathogenic yeasts in the genus trichosporon

The emergence of opportunistic and antifungal resistant strains has given rise to an urgent need for a rapid and accurate method for the detection of fungal pathogens. In this application, we demonstrate the detection of medically important fungal pathogens at the species level. The present method, which is based on a nucleotide hybridization assay, consists of a combination of different sets of fluorescent beads covalently bound to species specific capture probes. Upon hybridization, the beads bearing the target amplicons are classified by their spectral addresses with a 635 nm laser. Quantitation of the hybridized biotinylated amplicon is based on the fluorescent detection with a 532 nm laser. Using this technology we designed and tested various multiplex formats, the performance of forty eight species specific and group specific capture probes designed from sequence analysis in the D1/D2 region of ribosomal DNA, internal transcribed spacer regions (ITS), and intergenic spacer region (IGS). Species-specific biotinylated amplicons (>600 bp) were generated with three sets of primers to yield fragments from the three regions. The developed assay was specific and relatively fast, as it discriminated species differing by one nucleotide and required less than 50 min following amplification to process a 96 well plate with the capability to detect up to 100 species per well. The sensitivity of the assay allowed the detection as low as 102 genome molecules in PCR reactions and 107 to 108 molecules of biotinylated amplification product. This technology provided a rapid means of detection of Trichosporon species and had the flexibility to identify species in a multiplex format by combining different sets of beads. The assay can be expanded to include all known pathogenic fungal species.
Owner:MIAMI UNIVERISTY OF

Method for Determination of Minimum Inhibitory Concentration of Drugs

The invention discloses a method capable of rapidly, simply, conveniently and quantitatively detecting minimal inhibitory concentration, and the method comprises the following steps of: in accordance with the standard of CLSI (Clinical and Laboratory Standards Institute) (Version 2010), adding a fresh enterococcus suspension of a certain concentration into a sterile 96-well plate containing concentration gradient antibacterials for co-incubation; upon the ending of the 4-hour incubation, adding a fixed amount of fluorescent dyes SYTOX Green and DAPI (4,6-diamino-2-phenyl indole) to all the wells, protecting the wells from light for 15 minutes at room temperature, reading the fluorescent intensity of the two dyes in the wells by use of a fluorescent microplate reader respectively; after relevant background fluorescence is deducted, drawing a corresponding curve between bacterial fluorescence intensity ratio (Pdead/livel) and concentration of drug (CDrug), and determining the minimal concentration of drug corresponding to the moment the Pdead/livel is no longer fluctuated as the MIC (Minimal Inhibitory Concentration) of the antibiotic to the bacterium. The detection method provided by the invention has the characteristics of being simple, convenient, rapid and objective and being capable of performing mathematic statistics and analysis directly on detected data, becomes a new detection method for determining the minimal inhibitory concentration of antibacterials, and has a wide application prospect.
Owner:BEIJING FRIENDSHIP HOSPITAL CAPITAL MEDICAL UNIV

Improved expansion culture medium for regulatory T cells of human cord blood origin and application method of expansion culture medium

The invention relates to an improved expansion culture medium for regulatory T cells of human cord blood origin and an application method of the expansion culture medium. According to the expansion culture medium, heparin anticoagulated autologous cord blood plasma accounting for 10%-12% of the volume of a culture medium, CD3-CD28 antibody co-expressed immunomagnetic beads, recombinant human interleukin 2, 2-mercaptoethanol, rapamycin, HEPES (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid) and gentamicin are added into the RPMI (Roswell Park Memorial Institute)1640 culture medium; and then separated cell suspension is inoculated in a 96-well plates with a U-shaped bottom, hole-division expansion can be performed every 1-2 days, and the expansion period is 3-4 weeks. All reagents in the culture system reach the GMP (good manufacturing practice) level or are originated from autologous cord blood, so that risks caused by ingredients of animal origin are avoided, and the regulatory T cells can be used for a third-party unrelated donor and directly applied to clinical disease treatment; and compared with a traditional culture system, Treg cells (the regulatory T cells) expanded by the improved culture medium is excellent in aspects of growth speed, purity, activity, lymphocyte inhibition function and the like, and the Treg cells are expected to be used as the regulatory T cells of the third-party unrelated donor and applied to the clinical disease treatment.
Owner:HUNAN XENO LIFE SCI

Screening method of high flux 96 orifice plate for herbicide

The utility model provides a screening method used for herbicide high flux 96-hole orifice plate. The utility model adopts the technical proposal that: firstly, exponential phase chlorella is inoculated on the 96-hole orifice with a culture medium which is suitable for the chlorella, and the initial inoculating number of the chlorella is 7 to 8 x 105 per mL; secondly, the concentration gradient herbicide is dipped into each hole, and each concentration is provided with two to five duplicate samples as well as an empty control sample. The culturing condition is that: temperature is 25 plus or minus 0.2 DEG C, light is 2000Lx, continuous light is kept, fresh-keeping film is used to seal, no nutrient solution is added, chlorella solution is aerated four to five times every day, the chlorella is cultivated 72 to 144 hours in total, test sample is measured at 630nm absorbance every day, and then the EC50 (median effect concentration) of the herbicide on the chlorella is calculated according to the relationship between the absorbance and the herbicide concentration. Compared with the flask method, the utility model has the advantages of reducing herbicide amount in screening, and having easy operation, fast speed and high sensitivity.
Owner:ZHEJIANG UNIV OF TECH
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