Detection of mast cell degranulation by fluorescent molecular diffusion dynamic measurement
10.3969/j.issn.1000-4718.2014.09.031
- VernacularTitle:荧光分子扩散动态测量在肥大细胞脱颗粒体外检测中的应用
- Author:
Yining SUN
;
Yanan SUN
;
Yunong WANG
;
Jinghai ZHANG
;
Shuhua MA
;
Yi WANG
- Publication Type:Journal Article
- Keywords:
Fluorescence correlation spectroscopy;
Raster image correlation spectroscopy;
Diffusion coeffi-cient;
Mast cell degranulation
- From:
Chinese Journal of Pathophysiology
2014;(9):1717-1723
- CountryChina
- Language:Chinese
-
Abstract:
AIM:To establish a rapid in vitro method for mast cell degranulation tracing by raster image corre-lation spectroscopy (RICS).METHODS:RBL-2H3, a basophilic granulocyte mast cell line transfected with CD 63-GFP plasmid, was used for evaluating the methods , including β-hexosaminidase ( HEX) colorimetric assay, scanning electron microscopy (SEM) and RICS in the detection of mast cell degranulation .The sensibilities of these methods were com-pared.RESULTS:The sensitivities of β-HEX colorimetric assay and SEM were 5 mg/L and 3.9 ×10 -2 mg/L, respec-tively.RICS detection showed obvious decrease in the diffusion coefficient at dose of 3.9 ×10 -2 mg/L.CONCLUSION:Fluorescent molecular diffusion dynamic measurement can be used for rapid tracing of allergic substances in vitro.Accord-ing to the results, RICS can achieve nearly the same extent of sensitivity as the SEM does and is far more sensitive than β-HEX colorimetric assay.Compared with SEM, RICS has several advantages: it is faster, simpler and cheaper; it can be used in living cells;it is more suitable for rapid in vitro allergenic compounds tracing .Therefore, RICS is applicable in clinic allergic antigen screening and may also be used in pharmaceutical quality control .
