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- W2354314765 abstract "【Objective】 To evaluate the immunostaining effect of three different fixatives by confocal microscopy. 【Methods】 NRK-52E cells were cultured and fixed by 4 g / L PFA, methanol, and acetone, respectively, followed by three different antibodies to determine the representative antigens in membranes, cytoplasm, and nucleus under the confocal microscopy. Primary synovial fiber-like cells were cultured and fixed by 4 g / L PFA, permeabilizing by TritonX-100 and cooled methanol respectively. Immunostaining results of detecting the P-ERM protein were performed by confocal microscopy under the same observing condition. 【Results】 The signals of a cytoplasm skeleton protein and α-smooth muscle actin (α-SMA) showed sharp and clear, when the NRK cells were treated with acetone. A membranes protein, ZO-1 showed best results when the cells were treated with methanol. Compared to other fixatives, the cells fixed with 4 g / L PFA showed the best co-localization results of p-Smad3 protein with nucleus signals. The signals of P-ERM on primary synovial fiber-like cells indicated that it was significantly better when the cell permeabilizing by cooled methanol, comparing with permeabilization by TritonX-100. 【Conclusion】 Organic fixative were applied for fixing the cytoskeleton components and cross-linking fixative were suit for fixing the membrane components. Fixing by cross- linking agents followed by permeabilization of organic solvents can keep the antigens well and therefore could be used to take high quality pictures." @default.
- W2354314765 created "2016-06-24" @default.
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- W2354314765 date "2009-01-01" @default.
- W2354314765 modified "2023-09-25" @default.
- W2354314765 title "Evaluation of Different Fixatives in Confocal Immunofluorescence" @default.
- W2354314765 hasPublicationYear "2009" @default.
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