ChemoMetec NucleoCounter, Руководство пользователя

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6 Total and viability count
31
Cell sample Lysis buffer
Stabilizing
buffer
Multiplication
factor
200 µl 200 µl 200 µl 3
200 µl 400 µl 400 µl 5
200 µl 600 µl 600 µl 7
200 µl 800 µl 800 µl 9
When it is desired to dilute the stabilized lyzate it is
recommended to mix equal portions of Lysis buffer, Stabilizing
buffer and cell culture media (optionally a balanced salt
solution such as PBS) and then dilute the stabilized lyzate
with the mixture.
It is also possible to dilute the cell sample prior to
treatment with Lysis buffer and Stabilizing buffer using either
cell culture media or a balanced salt solution. When using this
method to dilute a cell suspension, take out a sample of the
diluted cell suspension and add Lysis buffer and Stabilizing
buffer in the normal way. Finally calculate the multiplication
factor.
Example of multiplication factor calculation:
The cell suspension is diluted 5 times by taking out 200 µl and
adding 800 µl PBS. Then a 200 µl sample is drawn from the
diluted suspension, 200 µl Lysis buffer and 200 µl Stabilizing
buffer is added. Thereby the sample is diluted another 3 times.
The cells are counted on the NucleoCounter.
The multiplication factor is calculated from the two separate
dilutions. In this example the multiplication factor is 5x3 =
15. Therefore the concentration of cells obtained from the
NucleoCounter has to be multiplied by 15 to calculate the
concentration of cells in the cell suspension.