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Microfluorimetric analysis of spatial and temporal patterns of immobilized cell growth
Author(s) -
Kuhn Robert H.,
Peretti Steven W.,
Ollis David F.
Publication year - 1991
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.260380404
Subject(s) - biological system , dna , biophysics , chemistry , cell , cell growth , microfluidics , chromatography , biology , nanotechnology , biochemistry , materials science
Pronounced spatial nonuniformities in cell density, physiology, and activity frequently arise within densely packed immobilized cell supports. For a more fundamental understanding of immobilized cell phenomena, we have developed high‐resolution microfluorimetric procedures to analyze local variations in both immobilized cell loading and growth rate. Fluorescent staining of total cellular DNA provides a measure of local biomass density. Actively growing (DNA synthesizing) cells are marked by pulse‐labeling newly synthesized DNA with the thymine analog, bromouracil. An immunofluorescent technique allows subsequent detection of spatial variations in DNA synthesis rates. These procedures enable the influence of mass‐transfer limitations and other immobilization effects on cell distribution and activity to be readily quantified. We demonstrate this approach through analysis of the patterns of growth of Escherichia coli entrapped within Sr‐alginate gel beads. The experimental techniques are potentially applicable to a variety of other aggregate cell systems.