Continuous and long-term volume measurements with a commercial Coulter counter

• Published in: PloS one Vol. 7; no. 1; p. e29866
• Main Authors: Bryan, Andrea K, Engler, Alex, Gulati, Amneet, Manalis, Scott R
• Format: Journal Article
• Language: English
• Published: United States Public Library of Science 17.01.2012; Public Library of Science (PLoS)
• Subjects: Adaptation; Algorithms; Animals; Aperture; Apoptosis; Baking yeast; Bioengineering; Biology; Cancer; Cell Biology - instrumentation; Cell culture; Cell Cycle; Cell Line, Tumor; Cell size; Control stability; Data processing; Deoxyribonucleic acid; DNA; Electrolytes; Engineering; Flow cytometry; G1 Phase; Growth rate; Homeostasis; Kinases; Medical research; Mice; Models, Biological; Population; Saccharomyces cerevisiae; Saccharomyces cerevisiae - classification; Saccharomyces cerevisiae - cytology; Saccharomyces cerevisiae - growth & development; Software; Species Specificity; Temperature control; Time Factors; United States > US; Massachusetts
• ISSN: 1932-6203; 1932-6203
• DOI: 10.1371/journal.pone.0029866

We demonstrate a method to enhance the time resolution of a commercial Coulter counter and enable continuous and long-term cell size measurements for growth rate analyses essential to understanding basic cellular processes, such as cell size regulation and cell cycle progression. Our simple modifications to a commercial Coulter counter create controllable cell culture conditions within the sample compartment and combine temperature control with necessary adaptations to achieve measurement stability over several hours. We also wrote custom software, detailed here, to analyze instrument data files collected by either this continuous method or standard, periodic sampling. We use the continuous method to measure the growth rate of yeast in G1 during a prolonged arrest and, in different samples, the dependency of growth rate on cell size and cell cycle position in arrested and proliferating cells. We also quantify with high time resolution the response of mouse lymphoblast cell culture to drug treatment. This method provides a technique for continuous measurement of cell size that is applicable to a large variety of cell types and greatly expands the set of analysis tools available for the Coulter counter.