Sam:LabNotes/Micro-manipulation/2011-4-12

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Revision as of 22:51, 12 April 2011 by >Sam Chiang (→‎E.coli cell microscopy analysis)
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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

File:Dapi stained dish-bf1.png File:Dapi stained dish-dapi200.png

File:Dapi stained ecoli-bf1.png File:Dapi stained ecoli-dapi100.png
 
File:No cells dish-bf1.png File:No cells dish-dapi200.png

File:No cells slide-bf1.png File:No cells slide-gfp500.png


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