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E. Coli glycerol stock cytometryModerator: BioTeam
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E. Coli glycerol stock cytometryI have recently been trying to count E. coli using a Petroff-Hauser counting chamber. I wanted to count cells straight out of my glycerol stocks, but I am having difficulties.
The E. coli looks strange directly after the glycerol stock is thawed. My Procedure: Thaw the E. coli cell bank (glycerol stock) at 37 C. Dilute the cell bank at a 1:10 dilution in saline & pipette 50 uL into the counting chamber. View on scope at 400X magnification. Specifically, the cells look elongated and many appear to be fused together in a linear orientation. Often times, several appear to be fused in chains of 5 or 6 individual units. This is strange because if I view E. coli from a live and growing culture, I can see discrete cells. Can anyone explain to me what I am observing? Is this common? What causes it? Are there any handling precautions I can take to minimize this phenomenon?
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