Document Detail


Inline characterization of cell concentration and cell volume in agitated bioreactors using in situ microscopy: application to volume variation induced by osmotic stress.
MedLine Citation:
PMID:  11857283     Owner:  NLM     Status:  MEDLINE    
Abstract/OtherAbstract:
A new in situ microscope (ISM) was developed and tested to perform in-line monitoring of average cell volume and cell concentration in agitated cultures subjected to osmotic stress. The ISM is directly immersed into the agitated broth in a bioreactor and generates still images of cells by using pulsed luminescent diode illumination and a virtual probe volume defined by depth of focus. This technique allows the acquisition of microscopic still images without mechanical sampling techniques. The front end of the sensor fits into a standard 25-mm port and it can be steam sterilized together with the bioreactor. The automatic image evaluation generates signals of the cell concentration and the average cell volume with a time resolution of a few minutes per data point (if a 200 MHz PC is used). Without the need for evaluation, the images can be acquired and stored at a rate of one image per 0.6 s. Hansenula anomala was cultivated as batch fermentation and monitored inline with the ISM. The ISM signal of the cell concentration agreed well with referential growth curves that were obtained from counting with a hemocytometer. The ISM signal of the average cell volume shows a gradual volume reduction as a result of the aging of the culture, and it monitors an abrupt and strong cell contraction if osmotic shocks are generated in the bioreactor. Systematic in vitro studies of osmotic shocks were performed by applying the ISM to agitated culture samples of H. anomala. The volume signal of H. anomala during osmotic shocks showed a very fast cell contraction within less than a second. Within half an hour after the shocks, no signal drifts were observed, which would indicate volume restoration. These findings suggest that the ISM volume signal can be used as an inline indicator of osmotic stress in cell cultures.
Authors:
V Camisard; J -P Brienne; H Baussart; J Hammann; H Suhr
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Publication Detail:
Type:  Journal Article; Research Support, Non-U.S. Gov't    
Journal Detail:
Title:  Biotechnology and bioengineering     Volume:  78     ISSN:  0006-3592     ISO Abbreviation:  Biotechnol. Bioeng.     Publication Date:  2002 Apr 
Date Detail:
Created Date:  2002-02-21     Completed Date:  2002-09-06     Revised Date:  2006-11-15    
Medline Journal Info:
Nlm Unique ID:  7502021     Medline TA:  Biotechnol Bioeng     Country:  United States    
Other Details:
Languages:  eng     Pagination:  73-80     Citation Subset:  IM    
Copyright Information:
Copyright 2002 Wiley Periodicals, Inc. Biotechnol Bioeng 78: 73-80, 2002; DOI 10.1002/bit.10178
Affiliation:
Laboratoire d'Automatique et d'Informatique de Lille/Automatique & Biotechnologies Upres-A 8021, France.
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MeSH Terms
Descriptor/Qualifier:
Bioreactors*
Cell Count / methods*
Cell Division
Cells, Cultured / cytology
Image Processing, Computer-Assisted / instrumentation,  methods*
Microscopy, Video / instrumentation*,  methods*
Osmotic Pressure
Pichia / cytology,  growth & development
Reference Standards
Sensitivity and Specificity
Stress, Mechanical
Comments/Corrections
Erratum In:
Biotechnol Bioeng 2002 May 5;78(3):352

From MEDLINE®/PubMed®, a database of the U.S. National Library of Medicine


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