Microfluorimetric analysis of spatial and temporal patterns of immobilized cell growth.

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 Bioeng

authors

Kuhn RH,Peretti SW,Ollis DF

doi

10.1002/bit.260380404

subject

Has Abstract

pub_date

1991-08-05 00:00:00

pages

340-52

issue

4

eissn

0006-3592

issn

1097-0290

journal_volume

38

pub_type

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