Magnafire ccd camera

WT and TMEM10 Oli-sneu cells were cultured under proliferation conditions for 48 hrs. Cells were then fixed with 4% PFA and stained with Cell Tracker dye (ThermoFisher). Images were taken with an ImageXpress microscope (Molecular Devices) and analyzed using MetaXpress software. Measurements taken included process outgrowth; number of branches per cell; number of processes per cell; and percentage of cells exhibiting process outgrowth. For morphological analysis of primary OPCs, cell cultures were fixed in 4% PFA and immunolabeled for MBP. Images were taken using an Axiovert 100 microscope (Carl Zeiss) with a MagnaFire CCD camera (Optronics). 50–100 cells were randomly selected and individually outlined using the selection tool in ImageJ^{41 (link)}. Mean fluorescence intensity and cell area were measured and multiple shape descriptors calculated using ImageJ. Circularity is defined as the degree to which a particle is similar to a circle, taking into consideration the smoothness of the perimeter and solidity as an inverse measure of the overall concavity of a particle⁴² .

T. vaginalis cells in logarithmic growth phase were washed and fixed with 4% paraformaldehyde in Phosphate Buffered Saline (PBS) pH 7.2, for 1 h. Cells were then washed three times in PBS, spotted on a slide and permeabilized with ice-cold methanol for 2 min. Slides were then incubated for 1 h with an anti-TvSaplip12 monoclonal antibody, obtained as previously described (Cuccuru et al., 2012 (link)).
After three washes in PBS-Tween 20 0.1%, slides were incubated with fluorescein isothiocyanate (FITC)-conjugated polyvalent anti-mouse antibody (Sigma-Aldrich) and 5 μg/ml 4′,6-diamidino-2-phenylindole (DAPI) for nucleus staining. An anti-Salmonella abortus ovis monoclonal antibody was used as negative control. Samples were observed using an Olympus BX51 microscope and the images were acquired with Optronics MagnaFire CCD Camera.