Introduction
The following protocol explains how to use the temperature controller CherryTemp with the Therma Flow chip for medium perfusion when yeast cells are mounted on lectin coated slides. Yeast cells are deposited on lectin coated coverslips and the Therma Flow chip is placed on top. Lectin helps to reduce cell movement for optimal imaging and CherryTemp allows for an accurate temperature control without disturbing cells. The Therma Flow chip allows for replacement of medium and drug injection.
Step-by-step guidelines:
Step 1: Pipet a drop of lectin (Concanavalin A, Con A) on a high-resolution 24x60mm glass coverslip.
Step 2: Let the liquid evaporate under the hood. Rinse with water and dry.
Step 3: Place the glass coverslip on the CherryTemp dedicated stage insert. Remove the sticker from the Therma Flow chip and paste it on the coated glass coverslip. The insert helps to keep coverslip and Therma Flow well aligned.
Step 4: Once theTherma Flow chip sticks to the glass coverslip (engraved arrows and luers facing up, glass surface and microfluidic chamber facing the coverslip) gently press with your fingers for proper attachment.
Step 5: Pipette the yeast sample in the inlet port. Prior concentration of cells by centrifuge spin is recommended to achieve an optimal cell density. Troubleshoot optimal density before pipetting.
Step 6: Use a syringe to vacuum medium and make yeast spread through the channel on the glass coverslip. Yeast will bind to the lectin-coated coverslip.
