Evaluation method
Set a centrifugal filtration plate on a conventional 96-well round bottom plate and pass cells through filter → centrifugation
Evaluation point: Presence or absence of contamination after plate centrifugation
Method (experimental condition)
Presence or absence of contamination
HEK293T not expressing GFP and HEK293T expressing GFP (human fetal kidney epithelial cell) are filtered in adjacent wells and judged by flow cytometry
Operation
① Cells were stripped with trypsin/EDTA and centrifuged.
② Cells were washed with PBS.
③ Cells were diluted with PBS to make about 2 x 104 cells/200 μL.
④ PI was added to make a final concentration of 0.25 μg/mL.
⑤ A centrifugal filtration plate was set on a 96-well plate.
⑥ Cells were added to wells in 200 μL.
⑦ The plate was centrifuged at 300 g x 30 s.
⑧ It was measured with flow cytometer (Guava easyCyte 8HT) .
*Measurement was made only within the red frame.
Operation conditions
It is being put in a centrifugal filtration plate
There was no suspension left in the filter.
After centrifugation
Results
Summary of verification results
・After centrifugation, there was no contamination in another well.
・This time, it was centrifuged at 300 g x 30 seconds, and all the suspension had fallen to the bottom.
