Microscopy raw and processed data to measure cell size, cell death and multi-nucleated ratio in yeast cells.
The data contains the raw data folder and a processed folder. The raw data is divided into Bright Field experiments and DAPI. The BrightField Microscopy images were taken on the strains bem1::KanMx pGal-Cdc42-sfGFP,bem1bem3::CloNAT pGal-Cdc42-sfGFP,pGal-Cdc42-sfGFP, and WT. The goal was to measure the cell sizes as a proxy of the polarization state for those strains in specific galactose concentrations taken by the population growth results. We use three galactose concentrations : 0%, 0.06%, and 0.1%. %=w/v
The processed folder contains an Excel file with three sheets: cell radii ratio, cell death radio,, and multinucleated cells ratio with the post-processed relevant variables from the raw data.
All microscopy was performed with a Nikon Eclipse Ti-E inverted microscope with an oil immersion 60x objective,1.40 of numerical aperture, and a refractive index of 1.51. The software used for data collection was Nis-Elements Ar (https://www.microscope.healthcare.nikon.com/products/software/nis-elements/nis-elements-advanced-research) version 4.51.
Cells that do not polarize are predicted to have nuclear divisions but no cellular division, and therefore, we hypothesize that cells with polarization defects are more likely to be multinucleate. We measured the percentage of multinucleated cells for the different genetic backgrounds and galactose concentrations using DAPI staining combined with fluorescence microscopy. The DAPI staining protocol was taken from doi: dx.doi.org/10.17504/protocols.io.eigbcbw. Samples were imaged in a cover slip right after the DAPI staining. We used a Nikon Eclipse Ti-E inverted microscope with an oil immersion 60x objective and 1.40 numerical aperture. Its refractive index is 1.51. We imaged in two channels, namely, Brightfield and DAPI Fluorescence. The emission wavelength was 450 nm, and the excitation wavelength was 395 nm. The exposure time was 70ms, and the laser power was 1\%. The software used for data collection was Nis-Elements Ar (https://www.microscope.healthcare.nikon.com/products/software/nis-elements/nis-elements-advanced-research) version 4.51. Data analysis was performed with Cell Counter[https://imagej.nih.gov/ij/plugins/cell-counter.html ] plugin from ImageJ, version 1.53t, which requires Java 1.8.0_172(64-bit).
Each cell was labeled to count the total number of cells in each frame. Further, each cell with two or more nuclei and the dead cells were differently labeled.
- 2023-07-17 first online
- 2023-09-13 published, posted