Concanavalin A mounting protocol
Introduction
The following protocol explains how to use the temperature controller CherryTemp when yeast cells are mounted on lectin coated slides. Yeast cells are deposited on lectin coated coverslips and our dedicated spacers allow to simply place the CherryTemp chip on top. Lectin helps to reduce cell movement for optimal imaging and CherryTemp allows for an accurate temperature control without disturbing cells.
No shear stress and an easy protocol for rapid mounting and imaging of yeasts.
Step-by-step guidelines:
Step 1: Pipet a drop of lectin (Concanavalin A, Con A) on a high-resolution glass coverslip.
Step 2: Let the liquid evaporate under the hood.
Step 3: Top the coverslip with our dedicated spacer.
Step 4: Drop the yeast sample (up to 15µl) centred in the spacer hole. Concentration of cells is recommended to achieve an optimal cell density.
Step 5:
Top with the CherryTemp chip (engraved arrows facing up, glass surface and microfluidic chamber facing the sample).
Step 6:
Place the mounted slide on either an upright or an inverted configuration by simply flipping it.
References
Methods adapted from P. Tran lab (Institut Curie) and from our user’s feedback.
