Thin films of Type 1 collagen for cell by cell analysis of morphology and tenascin-C promoter activity.
BACKGROUND
The use of highly reproducible and spatiallyhomogeneous thin film matrices permits automated microscopy and quantitative determination of the response of hundreds of cells in a population. Using thin films of extracellular matrix proteins, we have quantified, on a cell-by-cell basis, phenotypic parameters of cells on different extracellular matrices. We have quantitatively examined the relationship between fibroblast morphology and activation of the promoter for the extracellular matrix protein tenascin-C using a tenascin-C promoter-based GFP reporter construct.


RESULTS
We find that when considering the average response from the population of cells, cell area correlates with tenascin-C promoter activity as has been previously suggested; however cell-by-cell analysis suggests that cell area and promoter activity are not tightly correlated within individual cells.


CONCLUSIONS
This study demonstrates how quantitative cell-by-cell analysis, facilitated by the use of thin films of extracellular matrix proteins, can provide insight into the relationship between phenotypic parameters.
Langenbach
Kurt J
KJ
Biotechnology Division/National Institute of Standards and Technology, Gaithersburg, MD 20899, USA. klangenbach@stmarkdc.org
Elliott
John T
JT
Tona
Alex
A
McDaniel
Dennis
D
Plant
Anne L
AL
D000199:Actins / Q000378:metabolism; D000818:Animals; D002450:Cell Communication; D024042:Collagen Type I / Q000737:chemistry*; D003599:Cytoskeleton / Q000648:ultrastructure; D016326:Extracellular Matrix Proteins / Q000737:chemistry; D005347:Fibroblasts / Q000166:cytology* / Q000378:metabolism*; D005786:Gene Expression Regulation; D049452:Green Fluorescent Proteins; D051379:Mice; D041681:NIH 3T3 Cells; D011401:Promoter Regions, Genetic / Q000502:physiology*; D019063:Tenascin / Q000235:genetics*
D000199:Actins; D024042:Collagen Type I; D016326:Extracellular Matrix Proteins; D019063:Tenascin; D049452:Green Fluorescent Proteins