Cytation confocal imaging reader

For cytoskeleton staining, cells were incubated with Alexa Fluor 488 phalloidin (Thermo Fisher), a high-affinity filamentous actin (F-actin) probe conjugated to green-fluorescent Alexa Fluor 488 dye (Thermo Fisher) for 1 h. The stained cells were mounted with DAPI-containing ProLong Antifade Mountant (Thermo Fisher) and observed using a Cytation Confocal Imaging Reader (Biotek/Agilent). To quantitatively analyze fluorescent intensities, cells of interest and regions of interest (ROI) were selected and measured for mean fluorescence intensity using ImageJ software. Corrected Total Cell Fluorescence (CTCF) was calculated based on: CTCF = Integrated Density − (Area of Selected Cell × Mean Fluorescence of Background readings).

For cytoskeleton staining, cells were incubated with Alexa Fluor 488 phalloidin (Thermo Fisher), a high-affinity filamentous actin (F-actin) probe conjugated to green-fluorescent Alexa Fluor 488 dye (Thermo Fisher) for 1 h. The stained cells were mounted with DAPI-containing ProLong Antifade Mountant (Thermo Fisher) and observed using a Cytation Confocal Imaging Reader (Biotek/Agilent). To quantitatively analyze fluorescent intensities, cells of interest and regions of interest (ROI) were selected and measured for mean fluorescence intensity using ImageJ software. Corrected Total Cell Fluorescence (CTCF) was calculated based on: CTCF = Integrated Density − (Area of Selected Cell × Mean Fluorescence of Background readings).