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KMID : 0880220120500030419
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Journal of Microbiology
2012 Volume.50 No. 3 p.419 ~ p.425
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Evaluation of the cell growth of mycobacteria using Mycobacterium smegmatis mc2 155 as a representative species
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Jorge A. Gonzalez-y-Merchand
Jorge F. Cerna-Cortes
Sandra Rivera-Gutierrez
Addy C. Helguera-Repetto
Rosalina Guadarrama-Medina
Ruben Zaragoza-Contreras
Leopoldo Santos-Argumedo
Robert A. Cox
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Abstract
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The study of the in vitro cell growth of mycobacteria still remains a fastidious, difficult, and time-consuming procedure. In addition, assessing mycobacterial growth in the laboratory is often complicated by cell aggregation and slow growth-rate. We now report that the use of a stainless steel spring in the culture led to an absence of large cell clumps, to a decrease of dead cells in the exponential phase and to growth of a more homogeneous population of large cells. We also report that flow cytometry is a rapid, simple and reliable approach to monitor mycobacterial cell growth and viability. Here, we monitored Mycobacterium smegmatis cellular growth by optical density, dry cell mass, and colony forming units; in addition, viability, cell size and granularity profiles were analyzed by flow cytometry, and cell morphology by electron microscopy. Cultures monitored by flow cytometry may lead to a better understanding of the physiology of mycobacteria. Moreover, this methodology may aid in characterizing the cell growth of other fastidious species of microorganisms.
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KEYWORD
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mycobacterial growth, mycobacterial physiology, Mycobacterium smegmatis, flow cytometry
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