I am planing to screen my cell cultures for mycoplasma contamination using the Hoechst 33258 DNA staining method.
This method is suitable for adherent cultures. Is this method suitable for suspension cultures?
If yes, what is the procedure to attach my suspension culture on coverslip and prevent washout of cells during staining.
If you are working with suspension cells, it's always easier to use Flow cytometry but if you want to use microscopy then I would suggest using chamber slides. You just have to put your cell suspension inside the chamber and then stick the cells to the microscope slide. After that you can do whatever treatment you want just by putting in the chamber the proper solution and wash afterwards. The more the steps the more cells you'll lose mostly because of pipetting so try to pipette always at the same corner and be gentle. To stick the cells to the slide your best option is to centrifuge the slide. Unfortunately in my lab that wasn't an option so I just fixed the cells with methanol while there were suspended in PBS and it worked very well. Most of the cells were attached to the slide and the staining was perfect. Hope I helped. Good luck!
Contamination detection in suspension cells
For contamination detection in suspension cells, use the culture media of these cells: (with no antibiotic) NRK, 3T6, A549, VERO Cells These cells are good hosts, especially for Mycoplasma then contamination is easily detectable using optical microscopy after staining. check host cell culture for contamination before transferring suspension cells.
