In order to perform a fluorescence experiment you need to se

In order to perform a fluorescence experiment, you need to seed a 4-well chamber slide with 60,000 cells in each well. To prepare for the cell count, you transferred 20 uL of cell suspension into 30 uL of trypan blue. You calculated 19, 12, 25, 29 and 11 cells in 5 large boxes using the Neubauer hemacytometer. Based on this information, what is the cell density (cells per mL) of your cell suspension? How much cell suspension will you need to transfer roughly 60,000 cells into to each well?

Solution

Total Number of cells in five large boxes are 19+12+25+29+11 = 96

cell dilution = Cell volume suspended in trypan blue/ Cell volume suspended in trypan blue + volume of trypan blue

20/20+30

20/50

2/5

cell density is given by

96*10000/5*2/5

=480000 cells per ml.

In order to seed 60000 cells per well we need to seed

60000*1000/480000=125 microlitre

Concentration =	Number of Cells x 10,000
Number of square x dilution