Sam:LabNotes/Micro-manipulation/2011-4-12
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Single E.coli cell micromanipulation test on 04-12-'11
Testing objectives
- Focus on the performance of fluorescently stained cells.
- Question 1: Can we see the same number of samples under the BF GFP/DAPI filters? (If not, what do we missed, E.coli or unknown particles?)
- Question 2: Do we observe flourescently stained particles in negative control (blank slide)
Cell resource
- Using old stored E.coli K-12 culture (4C, 1XPBS, March, 2010)
- Estimated concentration = 10^9 CFU/mL
E.coli stain (SYBR-safe / DAPI)
- Prepare dye working solution (10X)
- Add 1uL SYBR-safe(10,000X) with 50uL 1X TE in a 1.7mL tube wrapped in foil => 200X
- Add 10uL SYBR-safe(200X) with 190uL 1X TE in a 1.7mL Amber tube => 10X
- Transfer 2uL E.coli culture into the 200uL dye working solution
- Same method was prepared for DAPI staining.
E.coli cell microscopy analysis
- Challenges:
- Next: