An automatic monitoring system for epithelial cell height

This paper describes an automatic method to measure cell height (h) of epithelia grown as monolayers on transparent filter supports. Tissues are mounted in an Ussing-type chamber enabling solution exchange on both sides. The apical and basal side of the epithelial cells are marked with fluorescent b...

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Bibliographic Details
Published in:Pflügers Archiv 1993-10, Vol.425 (1-2), p.164-171
Main Authors: VAN DRIESSCHE, W, DE SMET, P, RASKIN, G
Format: Article
Language:English
Subjects:
Autoanalysis - methods
Biological and medical sciences
Cell Line
Cell Membrane Permeability
Cell Size
Epithelial Cells
Fluorescent Dyes
Fundamental and applied biological sciences. Psychology
Glycerol - pharmacology
Hypotonic Solutions
Image Processing, Computer-Assisted
Kidney - cytology
Kinetics
Microspheres
Sucrose - pharmacology
Vertebrates: urinary system
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Summary:This paper describes an automatic method to measure cell height (h) of epithelia grown as monolayers on transparent filter supports. Tissues are mounted in an Ussing-type chamber enabling solution exchange on both sides. The apical and basal side of the epithelial cells are marked with fluorescent beads. The image of the fluospheres is captured with a video camera and processed by a computer-based video imaging system. One basal reference bead in a gelatin layer on the filter support and up to three beads attached at the apical surface are used to monitor changes in cell height of three cells simultaneously. The focusing of the microbeads is done automatically by moving the objective with a piezoelectric device mounted on the nosepiece of the microscope. The algorithm for locating the bead is based on the changes in fluorescent light intensity emitted by the fluospheres. The method has an accuracy higher than 0.1 micron and a time resolution as low as 6 s if measurements are restricted to one bead at the apical side. The method was tested on artificial model systems and used to measure volume changes in renal cultured epithelia (A6) after exposing the serosal surface to hypotonic solutions and replacing cell-impermeable sucrose by an organic compound (glycerol) with a smaller reflection coefficient. Serosal hypotonicity elicited a rapid volume increase followed by regulatory volume decrease, whereas the organic compound replacement caused a steady increase in cell volume.
ISSN:0031-6768
1432-2013
DOI:10.1007/BF00374517