I'm hoping to use CypHer5 mono ester dye to label bacteria, then infect cells with the labeled bacteria. I need to do a fixation step before microscopy, and I was wondering if the dye will be affected by the fixation?
Thanks in advance
Becky
"CypHer5E is a red-excitable, pH-sensitive cyanine dye derivative that is minimally fluorescent at a basic pH and maximally fluorescent at an acidic pH. It is therefore well suited to report the movement of a receptor from the cell surface into acidic endosomes upon agonist stimulation." from GE HealthCare, or what.
In normal microscopy, you will not see anything, because the general mounts are basic solutions (or it is not true, see later). You need live data acquisition, i am sure. Problem: your cellular acidic compartments will loss pH after fixation and processing. Another issue (see above): bacteria tends to have acidic surface... this issue looks problematic for me.
Hi Becky,
I am trying to do a similar assay... Could you do it?
Is Cypher5e-labeled cells/bacteria stable after 1 week or so?
Thank you so much!
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