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Value defining method for microsphere fluorescent intensity standard substance

A standard substance, fluorescence intensity technology, used in fluorescence/phosphorescence, measuring device, suspension and porous material analysis, etc., can solve the problem of inability to guarantee the accuracy and reliability of the instrument's measurement value, and improve the accuracy and reliability. Effect

Inactive Publication Date: 2014-10-15
NAT INST OF METROLOGY CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

At present, there are no relevant measurement standards and verification procedures in China, and the accuracy and reliability of the instrument's measurement value cannot be guaranteed.

Method used

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  • Value defining method for microsphere fluorescent intensity standard substance
  • Value defining method for microsphere fluorescent intensity standard substance
  • Value defining method for microsphere fluorescent intensity standard substance

Examples

Experimental program
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Effect test

Embodiment 1

[0039] A method for determining the value of a standard substance of microsphere fluorescence intensity, comprising the steps of:

[0040] (1) Prepare microsphere fluorescence intensity standard substance;

[0041] (2) Determining the purity of the fluorescein powder, in order to prepare the fluorescein standard mother solution for calculation;

[0042] (3) Calibrate the fluorescence spectrophotometer, prepare the fluorescein standard mother solution, and establish the calibration curve between the fluorescence response value of the instrument and the fluorescein concentration;

[0043] (4) Measure the particle number concentration of the microsphere fluorescence intensity standard substance: dilute the microsphere fluorescence intensity standard substance with PBS solution to obtain a suspension, draw the suspension with a pipette and place it on the grid filter, and Carry out suction filtration, intercept all the particles in the suspension on the grid filter, then count th...

Embodiment 2

[0051] A method for determining the value of a standard substance of microsphere fluorescence intensity, comprising the steps of:

[0052] (1) Preparation of microsphere fluorescence intensity standard substance:

[0053] Prepare isopropanol and distilled water according to the weight ratio of 0.2-2.0 to prepare isopropanol aqueous solution, then add 5g-15g vinyl acetate monomer, 0.5g-2.0g polystyrene base to 100g of the isopropanol aqueous solution Pyrrolidone and 0.1g-0.3g potassium persulfate react at 40°C-80°C for 10h, then dry at 80°C to obtain microspheres; weigh 1g-10g of the microspheres and suspend them in a 300g mass In NaOH methanol solution with a fraction of 5% to 20%, hydrolyze at 20°C to 80°C for 5 hours, then wash with distilled water, and then dry at 80°C to obtain microspheres with a nominal value of 7 μm carrying hydroxyl groups on the surface; Measure 0.1g to 5g of the microspheres carrying hydroxyl groups on the surface and add them to 50ml to 1000ml of f...

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Abstract

The invention discloses a value defining method for a microsphere fluorescent intensity standard substance. The method comprises the following steps: (1) preparing a microsphere fluorescent intensity standard substance; (2) measuring purity of fluorescein powder to calculate and prepare fluorescein standard mother liquid; (3) regulating a fluorospectro photometer; creating a calibration curve of an instrument fluorescent responding value and fluorescein density; (4) measuring granular quantity density of the microsphere fluorescent intensity standard substance; (5) measuring fluorescent responding value of a sample; calculating fluorescein density of the sample and fluorescent molecule quantity MESF carried by each microsphere fluorescent intensity standard substance granular. The value defining method is provided with favorable accuracy, reliability and traceability of value.

Description

technical field [0001] The invention relates to a method for determining the value of a standard substance, in particular to a method for determining the value of a standard substance for the fluorescence intensity of microspheres. Background technique [0002] Flow cytometry (Flow cytometry, FCM) is to irradiate single cells or particles stained by fluorescent pigments under high-speed flow state with high-energy laser, and measure the intensity of scattered light and emitted fluorescence, so as to analyze the cell (or particle) A modern cell analysis technology for qualitative or quantitative detection of physical, physiological, biochemical, immune, genetic, molecular biological traits and functional states, which can accurately quantitatively analyze and classify individual cells. [0003] Flow cytometry analysis has broad application prospects in basic medical, clinical and scientific research. In the absolute quantitative analysis of flow cytometry, the quantitative f...

Claims

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Application Information

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IPC IPC(8): G01N21/64G01N15/06G01N24/08
Inventor 刘俊杰修宏宇张伟
Owner NAT INST OF METROLOGY CHINA
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