Abstract:
: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.
journal_name
Biotechnol Bioengjournal_title
Biotechnology and bioengineeringauthors
Kuhn RH,Peretti SW,Ollis DFdoi
10.1002/bit.260380404subject
Has Abstractpub_date
1991-08-05 00:00:00pages
340-52issue
4eissn
0006-3592issn
1097-0290journal_volume
38pub_type
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journal_title:Biotechnology and bioengineering
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