Phenotypic differences between interfertile Chlamydomonas species - Cell Wall Analysis

This repository contains raw .TIF image data of Chlamydomonas reinhardtii (cc-124) and Chlamydomonas smithii (cc-1373) stained with Calcofluor White with Evan's blue, fixed in 4% PFA, and imaged on a spinning disk confocal microscope. The protocol to generate this data is described in the associated publication, "Phenotypic differences between interfertile Chlamydomonas species". 

Cells grown in TAP media were collected through centrifugation at 2,000 RPM for 1 minute. Cells were suspended in TAP media with the noted concentration of Calcofluor White (CFW) stain (Sigma-Aldrich, SKU 18909) for 10 minutes while rotating at ambient temperature. Afterward, cells were washed twice in fresh TAP media. For initial protocol development, cells were immediately imaged after washes. For the datasets quantified in this pub, cells were fixed in freshly made 4% paraformaldehyde in PBS for 15 minutes at ambient temperature while protected from light. Fixed cells were washed twice in PBS and stored at 4 C until imaged. 

We acquired micrographs with a Yokogawa CSU W1-SoRa scanner unit attached to a Nikon Ti2-E confocal microscope. We used a Plan Apo λ 40× air objective to collect images using standard DAPI channel settings. We acquired images at the medial focal plane of the cells in the field of view.

File Name Description: 
[well position]_[strain number]_[Treatment]_[prep]_[microscope]_[secondary magnification]_[image number]

The code to analyze these images is available on GitHub