as regards that etidum bromide stain is very sensitive to light
You can store only the unstained comet assay slides in a black box with cotton wet pad. When you stain the slides with any fluorescent dyes, it is imperative to observe, visualize and capture the comet images immediately. As the time exceeds, the intensity of the dye degrades and you wont get any appreciable results.
Spectrophotometric MTT assay
3-[4,5-dimethylthiazol-2-yl]-2,5-diphenyl tetrazolium bromide (Sigma-Aldrich) was dissolved in RPMI 1640 medium, at a concentration of 5 mg/ ml and fi ltered through a 0.2 μm filter. Subsequently, 100 μl of the yellow MTT solution was added to each well of a well plate, containing 1 ml of the cell suspension. The cells were then incubated at 37°C with 5% CO2. A blank solution was prepared according to the above procedure using complete medium without cells. After a three-hour incubation period, the resulting formazan crystals were dissolved using 1 ml of acidified isopropanol (0.05 N HCL in absolute isopropanol), and the absorbance of the obtained solution was measured at a wavelength of 570 nm using a spectrophotometer .
Hi.
Yes, it is possible.
I also use EtBr. Although, after a while, a few days/weeks, the cells are not visible when viewed, you only need to stain again with a little quantity (say 10ul of 10ug/ml EtBr) and your cells are clear for viewing.
This is my experience.
All the best.
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