Sack: Cell Harvesting for Electrophysiology: Difference between revisions
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(New page: Cell Plating for Electrophysiology<br> Jon Sack Feb 3, 2011<br> Ken Eum April 12, 2011 (Edit)<br> Notes: Steps should be done (not necessary if cell stock is only being used for plating c...) |
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Cell Harvesting for Electrophysiology<br> | |||
Ken Eum July 18, 2012 (edit)<br> | |||
Procedures (in 35mm dish): | Notes: Steps should be done (not necessary if cell stock is only being used for harvesting cells for electrophysiology) inside of a Biological Safety Cabinet in Sterile conditions. | ||
# Check under microscope to make sure that cells are | |||
# Carefully remove (aspirate) the solution from the | Procedures (in 35mm dish): | ||
# Rinse with | # Check under microscope to make sure that cells are ~80% confluent | ||
# Remove (aspirate) the | # Carefully remove (aspirate) the solution from the 35mm dish containing the cells | ||
# Add | # Rinse with Versene ~ 2 mL | ||
# Remove (aspirate) the Versene solution | |||
# Add 2 mL of Versene to detach cells | |||
# | # Wait ~1-2 minutes and scrape off cells with a cell scraper | ||
# | # Transfer the suspended cells into a 15mL conical tube | ||
# Add | # Add 3mL of CHO-SFM – II + 25mM HEPES + 1% Pen/Strep into the 15mL conical tube with the suspended cells. | ||
# Spin at 500rcf for 2 minutes | |||
# | # Aspirate out the supernatant | ||
# | # Resuspend with 2mL of CHO-SFM-II + 25mM HEPES | ||
# | # Transfer the suspended cells into a 2mL tube | ||
Latest revision as of 16:10, 15 January 2013
Cell Harvesting for Electrophysiology Notes: Steps should be done (not necessary if cell stock is only being used for harvesting cells for electrophysiology) inside of a Biological Safety Cabinet in Sterile conditions. Procedures (in 35mm dish):
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