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:

http://probes.invitrogen.com/media/pis/mp10078.pdf

Written by Zac Robinson, Dec 2010