Team:EPF-Lausanne/Protocol/TrypanBlue

From 2012.igem.org

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=== Sampling ===
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{| class="wikitable" style="text-align: center; color: black;"
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|Cell Density
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|Dilution
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|PBS
 +
|Cells
 +
|Trypan Blue
 +
|-
 +
|1-2
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|4
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|100
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|50
 +
|50
 +
|-
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|2-4.5
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|8
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|125
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|25
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|50
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|-
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|4.5-7
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|12
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|120
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|15
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|45
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|-
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|>7
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|16
 +
|137.5
 +
|12.5
 +
|50
 +
|-
 +
|}

Revision as of 16:40, 16 September 2012

Protocol: Trypan Blue Method


Sampling
Cell density (10^6 ml) Dilution PBS (µl) Cells (µl) Trypan Blue (µl)
1-2 4 100 50 50
2-4.5 8 125 50
4.5-7 12 120 15 45
>7 16 137.5 12.5 50


Sampling

Cell Density Dilution PBS Cells Trypan Blue
1-2 4 100 50 50
2-4.5 8 125 25 50
4.5-7 12 120 15 45
>7 16 137.5 12.5 50


  1. Take xµL ofPBS in 96 well plate
  2. Add required volume of cell culture. Mix once
  3. Bring the plate back to microscope, add trypan blue to the PBS + Cell mixture just before counting the sample. 
     Trypan Blue is toxic to cells. If left too long with trypan blue, even healthy cells will die.


Calculation of LCD :

LCD = Cell Count/ ( 100* 4) * Dilution

Tips :

  1. Mix cells before sampling
  2. Take cell sample from top of the liquid
  3. Mix trepan blue into the PBS + cel solution slowly and well before loading


Wiki Links

Important pages

Pages

Retrieved from "http://2012.igem.org/Team:EPF-Lausanne/Protocol/TrypanBlue"