Adapted from the lab compendium Snowflake Yeast Protocols — Ratcliff Lab / MuLTEE (V2).
- Follow the 5-day re-acclimation protocol (Culturing & settling selection › Daily growth and settling selection and Freezer stocks › Reviving strains from frozen stock).
- On the final day, transfer 100 μL of culture into fresh YPD and incubate for 12 h.
- Stain with Calcofluor White (5 μM final), 30 min in the dark.
- Image at ×40 objective, UV excitation of the blue fluorescent cell wall stain (Nikon Ti-E system).
Image analysis pipeline (cell segmentation): zipped folder containing pipeline is attached — Cellpose-based segmentation of individual cells, measures aspect ratio (AR) directly along with area, major/minor axis, circularity, and solidity.