Sam:LabNotes/Micro-manipulation/2011-4-12: Difference between revisions

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>Sam Chiang
>Sam Chiang
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==E.coli cell microscopy analysis==
==E.coli cell microscopy analysis==
  [[File:dapi_stained_dish-bf1.png|400px]] [[File:dapi_stained_dish-dapi200.png|400px]]
  [[File:041211_dapi_stained_dish-bf1.png|400px]] [[File:041211_dapi_stained_dish-dapi200.png|400px]]
   
   
  [[File:dapi_stained_ecoli-bf1.png|400px]] [[File:dapi_stained_ecoli-dapi100.png|400px]]
  [[File:041211_dapi_stained_ecoli-bf1.png|400px]] [[File:041211_dapi_stained_ecoli-dapi100.png|400px]]
    
    
  [[File:no_cells_dish-bf1.png|400px]] [[File:no_cells_dish-dapi200.png|400px]]
  [[File:041211_no_cells_dish-bf1.png|400px]] [[File:041211_no_cells_dish-dapi200.png|400px]]
   
   
  [[File:no_cells_slide-bf1.png|400px]] [[File:no_cells_slide-gfp500.png|400px]]
  [[File:041211_no_cells_slide-bf1.png|400px]] [[File:041211_no_cells_slide-gfp500.png|400px]]




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Revision as of 22:52, 12 April 2011

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:041211 dapi stained dish-bf1.png File:041211 dapi stained dish-dapi200.png

File:041211 dapi stained ecoli-bf1.png File:041211 dapi stained ecoli-dapi100.png
 
File:041211 no cells dish-bf1.png File:041211 no cells dish-dapi200.png

File:041211 no cells slide-bf1.png File:041211 no cells slide-gfp500.png


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