Optimization of yeast cell cycle analysis and morphological characterization by multispectral imaging flow cytometry

Area of Science:

• Cell Biology
• Microbiology
• Biophysics

Background:

• Budding yeast Saccharomyces cerevisiae is a key model for eukaryotic cell cycle studies.
• Conventional methods like visual analysis and flow cytometry have limitations in accuracy and scope.
• Accurate cell cycle analysis is crucial for understanding yeast biology and disease.

Purpose of the Study:

• To demonstrate multispectral imaging flow cytometry (MIFC) for precise yeast cell cycle and morphological phenotype quantitation.
• To compare MIFC with traditional visual and flow cytometry methods.
• To investigate the effects of NAP1 gene expression on yeast cell cycle and morphology.

Main Methods:

• Cell cycle analysis of wild-type, nap1Δ, and NAP1-overexpressing yeast.
• Comparison of visual analysis, conventional flow cytometry, and MIFC.
• Quantitative morphological analysis using algorithms for bud length and other features.

Main Results:

• MIFC provided reliable quantification of the yeast cell cycle, surpassing conventional methods.
• Observed G2/M delay and elongated buds in nap1Δ strains, consistent with prior findings.
• NAP1 overexpression led to elongated buds but only minor cell cycle disruption, suggesting independent regulation.

Conclusions:

• MIFC offers a powerful, accurate method for simultaneous cell cycle and morphological profiling in yeast.
• NAP1's impact on cell cycle and morphology appears independent, with varying expression levels yielding distinct phenotypes.
• This technique advances the study of cell cycle regulation and morphological changes in model organisms.