We need to normalize an experiment according to the relative number of cells, and we do not have a fluorescence microscope, so we were thinking of staining the cells nuclei with DAPI, lysate the cell and then quantify the fluorescence contained in the volume of the lysate.
It sounds like it might work, as long as you are confident that the treatment you are studying does not cause cell cycle arrest, which would affect the amount of DNA in the cells.
If you decide to try this method, you will have to validate it by comparing the readout to an actual cell number measurement, such as a hemacytometer count, or Coulter counter measurement. Of course, if you can make that measurement, you may not need the DAPI fluorescence measurement.
Dear Irais
Are your cells all mononuclear? In case they are not, then an indirect readout like DAPI fluorescence in lysate may not correlate well with the number of cells in your sample.
If cell count is all that you require and your cells can be detached or are in suspension, I would recommend to perform a basic trypan blue staining and counting on a hemocytomter.
Flow cytometry is a more reliable method (with beads to correct for cell count).
Good luck
Also, you may need to sonicate the DNA because it will clump together and your solution will not be well mixed.
You could always go to photomicroscopy and counting on ImageJ.
Thank you all for your answers and suggestions, you've been very helpful.
There are a range of live/dead toxicity kits from suppliers, using different methods. Some easily is applied in a lab. Both absorbance or fluo readout from plates. The choice is yours.
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