Method for dyeing fluorescent microballons

Abstract

This invention relates to a method for dyeing polymer microspheres to obtain fluorescent polymer microspheres. The method comprises: uniformly mixing fluorescein 0.01-80%, polymer microspheres 0.1-80%, emulsifier 0-10%, tackifier 0-10%, and solvent (one or more of good solvents and poor solvents) 18.9-99.89%, dyeing under 1-101 kPa in dark for 1-720 h, taking out the dyed polymer microspheres, and washing repeatedly. The obtained fluorescent polymer microspheres have such advantages as simple process, high fluorescent brightness, wide fluorescent spectrum range, uniform particle size distribution and low relative standard deviation, and can be used as absolute counting microspheres of flow cytometry, fluoroimmunoassay microspheres, biosensor, microfluidic chip, and calibrator of fluorescence microscope.

Description

Technical field [0001] The present invention relates to the technical field of analyzing and separating materials, in particular to a dyeing method of fluorescent microspheres. Background technique [0002] Flow cytometry analysis can be applied to many fields such as high-throughput analysis, metabolic process analysis, drug discovery, etc. This method has high sensitivity, selectivity and convenience. It can detect multiple parameters of a sample at the same time, and provide multi-faceted cellular information. However, in flow cytometry analysis, in order to accurately measure the number of cells, an absolute counting fluorescent microsphere is required as an internal reference for detection. The counting microspheres are required to have uniform particle size, uniform fluorescence intensity, and a wide fluorescence emission spectrum. At present, there are no domestic fluorescent microsphere products in the domestic market. The products are imported from abroad. Most of the fl...

AI Technical Summary

Problems solved by technology

At present, there are no domestic fluorescent microsphere products in the domestic market, and the products are all imported from abroad. Most of the fluorescent microspheres commonly used in flow cytometry come from InterfacialDynamics Corporation (IDC), Bangs Laboratories and other companies, and other foreign companies such as BD, Coulter, etc. also act as agents for fluorescence Microsphere absolute counting products, the domestic Jingmei company mainly acts as the agent of Bangs Laboratories products, and the reagents used by domestic hospitals at all levels and disease control centers in various provinces and cities are all products of foreign companies. The prices of these foreign reagents are very high, mainly due to the particle size and Fluorescent microsphere technology with uniform light intensity is cumbersome and complicated, so the cost remains high

[0003] Domestic patent CN1782020 invented silica fluorescent microspheres loaded with cadmium telluride fluorescent quantum dots, and patent CN1690163 used the induced phase transition method to obtain polymer-loaded quantum dot fluorescent microspheres, but due to the small particle size, it is difficult to be used in fluid flow Cytometer analysis; patent CN1693411 prepared magnetic fluorescent microspheres by spray drying method, but in actual operation, spray drying method is easy to inactivate fluorescein. However, the brightness of fluorescent microspheres is difficult to improve. Since the 1980s, there have been many patents related to the synthesis of fluorescent microspheres and the research on supporting reagents and instruments. However, because flow cytometry requires high uniformity of fluorescent microspheres, and the emission spectrum of fluorescent microspheres is required to detect multiple cell populations at the same time, how to prepare Fluorescent microspheres with low price, high brightness, uniform particle size and light intensity, and wide fluorescence emission spectral range have always been the focus of research in the field of flow cytometry.

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