Optimization of yeast cell cycle analysis and morphological characterization by multispectral imaging flow cytometry
Open Access
- 8 July 2008
- journal article
- research article
- Published by Wiley in Cytometry Part A
- Vol. 73A (9), 825-833
- https://doi.org/10.1002/cyto.a.20609
Abstract
Budding yeast Saccharoymyces cerevisiae is a powerful model system for analyzing eukaryotic cell cycle regulation. Yeast cell cycle analysis is typically performed by visual analysis or flow cytometry, and both have limitations in the scope and accuracy of data obtained. This study demonstrates how multispectral imaging flow cytometry (MIFC) provides precise quantitation of cell cycle distribution and morphological phenotypes of yeast cells in flow. Cell cycle analysis of wild‐type yeast, nap1Δ, and yeast overexpressing NAP1, was performed visually, by flow cytometry and by MIFC. Quantitative morphological analysis employed measurements of cellular length, thickness, and aspect ratio in an algorithm to calculate a novel feature, bud length. MIFC demonstrated reliable quantification of the yeast cell cycle compared to morphological and flow cytometric analyses. By employing this technique, we observed both the G2/M delay and elongated buds previously described in the nap1Δ strain. Using MIFC, we demonstrate that overexpression of NAP1 causes elongated buds yet only a minor disruption in the cell cycle. The different effects of NAP1 expression level on cell cycle and morphology suggests that these phenotypes are independent. Unlike conventional yeast flow cytometry, MIFC generates complete cell cycle profiles and concurrently offers multiple parameters for morphological analysis. © 2008 International Society for Advancement of CytometryKeywords
This publication has 55 references indexed in Scilit:
- A Dense Poly(Ethylene Glycol) Coating Improves Penetration of Large Polymeric Nanoparticles Within Brain TissueScience Translational Medicine, 2012
- Antibiotic Eluting Chitosan Glycerophosphate Implant in the Setting of Acute Bacterial Sinusitis: A Rabbit ModelAmerican Journal of Rhinology and Allergy, 2010
- Levodopa delivery systems: advancements in delivery of the gold standardExpert Opinion on Drug Delivery, 2010
- Advances in the treatment of neurodegenerative disorders employing nanotechnologyAnnals of the New York Academy of Sciences, 2009
- Computational Methods for Estimation of Cell Cycle Phase Distributions of Yeast CellsEURASIP Journal on Bioinformatics and Systems Biology, 2007
- Sensitivity measurement and compensation in spectral imagingCytometry Part A, 2006
- Distinguishing modes of cell death using the ImageStream® multispectral imaging flow cytometerCytometry Part A, 2004
- A model for the computer analysis of synchronous DNA distributions obtained by flow cytometryCytometry, 1980
- Cell cycle phase expansion in nitrogen-limited cultures of Saccharomyces cerevisiae.The Journal of cell biology, 1980
- Thymidine Kinase: Evidence for its Absence from Neurospora crassa and Some Other Micro-organisms, and the Relevance of This to the Specific Labelling of Deoxyribonucleic AcidJournal of General Microbiology, 1968