Hello everyone,
I am using mitotracker green to evaluate the total number of the mitochondria in primary cells after stimulation with different materials and i used Cytation 5 to measure the fluorescence intensity by area scanning, i used 48 well plate but several experiments i noticed the that the wells located in the middle section of the plate have always lower number comparing to the wells on the peripheral even for the same sample. Does any one face this before? or can any one tell me a valid way to quantify the mitochondrial number not using microscope?
I really appreciate your help in this.
have you normalized to cell numbers after fluorescence? (for example with Cristal Violet)? There might be less cells in the middle wells.
Dear Macarena Lucia Fernandez Carro , thanks for your reply. I used Hoechst to stain the nucleus of the cells the numbers seem very close so I assumed it's not the cell number problem.
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