In our thesis, we studied the PC-12 cell line, which represents a good neurological model for the research of neurodegenerative diseases. Analyses were performed using imaging flow cytometry. The ImageStream imaging cytometer combines conventional flow cytometry with modern fluorescence microscopy and allows multiparametric analyses of cell features, such as shape, texture, size, granularity and fluorescence intensity.
Cells were labelled with fluorophores (DAPI, Alexa Fluor 488, Alexa Fluor 633, TRITC) directly, with phalloidin or indirectly with antibodies. Using the IDEAS software, we determined the portion of live cells and the distribution of actin, cystatin C and cathepsin X.
With statistical analysis (determination of RD value) of phenotypic cell characteristics we studied subpopulations and individual cells. The fluorescence intensity was measured on permeabilised and non permeabilised cells. Higher signal strength was detected in permeabilised cells, as membrane permeabilisation allows labelled antibodies to enter into the cells.
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