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Comparative Physiology @ Guelph

Cellular Lights GFP-Tubulin Transduction on Silicon Strips

Protocol: Cellular Lights GFP-Tubulin Transduction on Silicon Strips

  • Materials
  • Prepare Cells
  • Transduction
  • D-PBS w/o Ca2+ and Mg2+
  • Cellular Lights GFP-Tubulin reagent
  • BacMam Enhancer
  • Dimethylsulfoxide (DMSO)
  • Compatible Cell Line (see manufacturers list)
  • Silicon Strip with 1 cm2 well
  • Seed cells in silicon wells ~24 hours prior to transduction at a density that results in a 70-80% confluent layer
  • Prepare 250 μL of Cellular Lights transduction solution by combining 90 μL of Cellular Lights reagent with 160 μL of D-PBS
  • Aspirate regular growth medium from the prepared cells and add diluted Cellular Lights transduction solution to the cells. Incubate the cells at room temperature in the dark for 2-4 hours with gentle rocking or shaking.
  • Aspirate Cellular Lights transduction solution from the plated cells and replace with appropriate culture medium with 1X BacMam Enhancer (1μL/mL). Incubate for 2 hours at 37°C and 5% CO2.
  • Aspirate enhancer solution and replace with regular cell growth medium. Incubate cells for >16 hours to allow for expression of the protein.
  • Transfer silicon strip to custom cell stretching device and mount on microscope.

Based on manufacturer’s protocol found at:

Written by Zac Robinson, Dec 2010