Calculate the number of cells/mL and how many uL are needed to have 25,000 cells.
If I count 70 cells in red square "B" and 64 cells in red square "C".
2squares(70+64)cells∗2=134 cells per red square
Here you divided by the number of squares counted to get the average cells per square. Then multiplied by 2 because of the 1:1 dilution in trypan blue.
We know the size of each red square is equivalent to 1 mm2 or 1000 um2
134∗104=1.34∗106 cells per 1000uL
Do a ratio to determine number of uL needed for 25K cells.
1000uL1,340,000cells=x25,000cells
Cross multiply to solve for x.
x=1,340,000cells(25,000cells)∗(1000uL)
This is the number of uL that has 25,000 cells.
x=18.66uL has 25,000 cells