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

From ZhangLabWiki
Jump to navigation Jump to search
>Sam Chiang
(Created page with '='''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…')
 
>Sam Chiang
Line 19: Line 19:


==E.coli cell microscopy analysis==
==E.coli cell microscopy analysis==
 
[[File:dapi_stained_dish-bf1.png|400px]]
 
*Challenges:




*Next:
*Next:

Revision as of 22:49, 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:Dapi stained dish-bf1.png


  • Next: