Abstract
In order to automate measurements of cell concentration and viability in a suspended animal cell culture, we have developed an in situ microscopic image analysis system with an effective cell recognition algorithm. With a small amount of sample, this system can measure the cell density rapidly and aseptically. In addition, it can measure a cell size histogram including cell debris small particle distribution. These small particles have been found to be related to the viability of the mouse-mouse hybridoma STK1 cell line. By using cell debris small particle density as an indicator of cell viability, the developed system provides non-destructive viability monitoring without trypan blue staining.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
References
Bradbury S (1979) Microscopical Image Analysis: Problems and Approaches. J. Microsc. 115: 137–150.
Broise Ddl, Noiseux M, Lemieux R & Massie B (1991) Long-Term Perfusion Culture of Hybridoma: A “Grow or Die” Cell Cycle System. Biotechnol. Bioeng. 38: 781–787.
Brooks RF & Shields R (1985) Cell Growth, Cell Division and Cell Size Homeostasis in Swiss 3T3 Cells. Exp. Cell Res. 156: 1–6.
Chuck AS & Palsson BO (1992) Population Balance between Producing and Nonproducing Hybridoma Clones Is very Sensitive to Serum Level, State of Inoculum, and Medium Composition. Biotechnol. Bioeng. 39: 354–360.
Frame KK & Hu W-S (1990) Cell Volume Measurement as an Estimation of Mammalian Cell Biomass. Biotechnol. Bioeng. 36: 191–197.
Goebel NK, Kuehn R & Flickinger MC (1990) Methods for Determination of Growth-Rate-Dependent Changes in Hybridoma Volume, Shape and Surface Structure during Continuous Recycle. Cytotechnol. 4: 45–57.
Livne A, Grinstein S & Rothstein A (1987) Volume-Regulating Behavior of Human Platelets. J. Cell. Physiol. 131: 354–363.
Miller SJO, Henrotte M & Miller AOA (1986) Growth of Animal Cells on Microbeads. I. In situ Estimation of Numbers. Biotechnol. Bioeng. 28: 1466–1473.
Pons M-N, Wagner A, Vivier H & Marc A (1992) Application of Quantitative Image Analysis to a Mammalian Cell Line Grown on Microcarriers. Biotechnol. Bioeng. 40: 187–193.
Resau JH & Trup BF (1988) Cell Injury, Differentiation, and Regeneration in Expiant, Organ, and Cell Culture Models. In: Advances in Cell Culture. Vol. 6 (pp. 261–289) Academic Press, Inc., London.
Rudt S, Blunk T & Muller RH (1992) Quantification of Cell Subpopulations, Fractions of Dead Cells and Debris in Cell Suspensions by Laser Difractometry. Pharm. Ind. 54: 966–969.
Sen S, Srienc F & Hu W-S (1989) Distinct Volume Distribution of Viable and Non-Viable Hybridoma Cells: A Flow Cytometric Study. Cytotechnol. 2: 85–94.
Shields R, Brooks RF, Riddle PN, Capellaro DF & Delia D (1978) Cell Size, Cell Cycle and Transition Probability in Mouse Fibroblasts. Cell 15: 469–474.
Tucker KG, Chalder S, Al-Rubeai M, Thomas CR (1994) Measurement of Hybridoma Cell Number, Viability, and Morphology Using Fully Automated Image Analysis. Enzyme Microb. Technol. 16: 29–35.
Wheatley DN, Inglis MS & Foster MA (1987) Hydration, Volume Changes and Nuclear Magnetic Resonance Proton Relaxation Times of HeLa S-3 Cells in M-Phase and the Subsequent Cell Cycle. J. Cell Sci. 88: 13–23.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1994 Springer Science+Business Media Dordrecht
About this chapter
Cite this chapter
Maruhashi, F., Murakami, S., Baba, K. (1994). Automated monitoring of cell concentration and viability using an image analysis system. In: Buckland, B.C., Aunins, J.G., Bibila, T.A., Hu, WS., Robinson, D.K., Zhou, W. (eds) Cell Culture Engineering IV. Current Applications of Cell Culture Engineering, vol 1. Springer, Dordrecht. https://doi.org/10.1007/978-94-011-0257-5_31
Download citation
DOI: https://doi.org/10.1007/978-94-011-0257-5_31
Publisher Name: Springer, Dordrecht
Print ISBN: 978-94-010-4114-0
Online ISBN: 978-94-011-0257-5
eBook Packages: Springer Book Archive