Abstract
A prototype of a newly developed apparatus for measuring cell growth characteristics of suspension cells in micro titre plates over a period of time was examined. Fully automated non-invasive cell counts in small volume cultivation vessels, e.g. 96 well plates, were performed with the Cellscreen system by Innovatis AG, Germany. The system automatically generates microscopic images of suspension cells which had sedimented on the base of the well plate. The total cell number and cell geometry was analysed without staining or sampling using the Cedex image recognition technology. Thus, time course studies of cell growth with the identical culture became possible. Basic parameters like the measurement range, the minimum number of images which were required for statistically reliable results, as well as the influence of the measurement itself and the effect of evaporation in 96 well plates on cell proliferation were determined. A comparison with standard methods including the influence of the cultured volume per well (25 μl to 200 μl) on cell growth was performed. Furthermore, the toxic substances ammonia, lactate and butyrate were used to show that the Cellscreen system is able to detect even the slightest changes in the specific growth rate.
Similar content being viewed by others
References
Berridge MV & Tan AS (1993) Characterization of the cellular reduction of 3-(4,5-dimethylthiazol-2-yl)2,5-diphenyltetrazolium bromide (MTT): Subcellular localization, substrate dependence, and involvement of mitochondrial electron transport in MTT reduction. Arch Biochem Biophys 303: 474-482.
Büntemeyer H, Lütkemeyer D & Lehmann J (1991) Optimization of serum-free fermentation processes for antibody production. Cytotechnology 5: 57-67.
Glauner B (1991) Viabilitätskontrolle in der Zellkulturtechnik. BioTec3 5: 58-61.
Gudermann F, Ziemeck P & Lehmann J (1996) CeDeX: Ein integriertes Analysesystem zur optischen Zelldichtebestimmung. Bioscope 4: 46-51.
Gudermann F, Ziemeck P & Lehmann J (1997) CeDeX: Automated Cell Density Determination. In: MJT Carrondo, B Griffith and JLP Moreira (eds) Animal Cell Technology from Vaccines to Genetic Medicine, Kluwer Academic Publishers, Dordrecht, pp. 301-307.
Jäger V, Lehmann J & Friedl P (1988) Serum-free growth medium for the cultivation of a wide spectrum of mammalian cells in stirred bioreactors. Cytotechnology 1: 119-129.
Mosman T (1983) Rapid colorimetric assay for cellular growth and survival: Application to proliferation and cytotoxicity sssays. J Immunol Meth 65: 53-63.
Tennant JR (1964) Evaluation of the trypan blue technique for determination of cell viability. Transplantation 2: 685-694.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Brinkmann, M., Lütkemeyer, D., Gudermann, F. et al. New technologies for automated cell counting based on optical image analysis `The Cellscreen'. Cytotechnology 38, 119–127 (2002). https://doi.org/10.1023/A:1021118501866
Issue Date:
DOI: https://doi.org/10.1023/A:1021118501866