At present I am studying biofilm and Bacteriophage interactions by using CLSM.many papers published with CLSM by using syto 9 and PI stains for study...Please suggest me any alternative stains for the above stains...
Please guide me....Research community ..
Thank you in advance...
Hi Pallavali,
I guess you are trying to assess how many cells are alive or dead in the biofilm? There are some alternatives to SYTO9 and PI, but they are mainly used for single-cell microscopy or for flow cytometry. You may need to check if they will work for biofilm. See below paper:
"Assessment of bacterial viability status by flow cytometry and single cell sorting"
DOI: 10.1046/j.1365-2672.1998.00436.x
In short, they use PI to assess permeabilized cells, EtBr to assess metabolically active cells (which exclude EtBr) and a bis-oxonol dye to assess membrane energetics. Instead of PI you might be able to use DAPI.
Hope this helps
Andrew
Hi, Pallavali,
I think I would need more information. Why won't SYTO 9 work for you? Are you just needing a different wavelength? Are you labelling all cells to get a total count? Live/dead assay? Wanting a label that isn't nucleic acid? Please specify and I may be able to help further.
Hi Jason,
Because of unavailability of syto 9, I asked for alternative dyes...luckely I got syto 9 for staining of biofilm and I got nice result with auto 9 and pi by CLSM...
Thank u...
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