# How to get enough cell alivefrom colon cancer tissue?

### #1

Posted 23 January 2001 - 10:00 PM

### #2

Posted 31 January 2001 - 10:00 PM

*prepare formazan

1-weight 5mg/ml of MTT ( pouder from sigma) using gloves and mask2-dissolve it in 25 ml of DMEM or your medium if different3-sterilise by filtering4-store at -20°C in black eppendorfs (MTT is Light degradable)

*Set up the multiwell plate

1-count celles in haemocytometer2-fill all the peripheric wells with sterile water3-fill the 1st column with medium only (It is the background)4-set up the plate with the right number of cells (it depends on your cells) add medium (100µl total max per well)5-incubate untill 80-100% confluence (40h)6-add 100 µl of MTT per well in the dark7-incubate 4 h8-add 100 µl DMSO9-read at least one hour after at 540 and 690 nm(ref)

*Calculations

1-do the mean for each column2-substract the mean of the backgroundcalculate the standard deviation and the standard error of the mean.

### #3

Posted 31 January 2001 - 10:00 PM

*prepare formazan

1-weight 5mg/ml of MTT ( pouder from sigma) using gloves and mask2-dissolve it in 25 ml of DMEM or your medium if different3-sterilise by filtering4-store at -20°C in black eppendorfs (MTT is Light degradable)

*Set up the multiwell plate

1-count celles in haemocytometer2-fill all the peripheric wells with sterile water3-fill the 1st column with medium only (It is the background)4-set up the plate with the right number of cells (it depends on your cells) add medium (100µl total max per well)5-incubate untill 80-100% confluence (40h)6-add 100 µl of MTT per well in the dark7-incubate 4 h8-add 100 µl DMSO9-read at least one hour after at 540 and 690 nm(ref)

*Calculations

1-do the mean for each column2-substract the mean of the backgroundcalculate the standard deviation and the standard error of the mean.