Functional proteometrics for cell migration

Feimo Shen, Louis Hodgson, Andrew Rabinovich, Olivier Pertz, Klaus Hahn, Jeffrey H. Price

Research output: Contribution to journalArticle

26 Scopus citations

Abstract

Background: Advances in living cellular fluorescence biosensors and computerized microscopy enable a vision of fully automated high-resolution measurements of the detailed intracellular molecular dynamics directly linked to cellular behaviors. Given the heterogeneity of cell populations, a statistically relevant study of molecular-cellular dynamics is a key motivation for improved automation. Methods: We explored automating computerized, microscope-based data extraction and analyses that monitor cell locomotion, rates of mitoses, and spatiotemporal activities of intracellular proteins via ratiometric fluorescent biosensors in mouse fibroblasts. Novel image processing methods included K-means clustering segmentation preprocessing followed by modified discrete, normalized cross-correlational alignment of two-color images; ratiometric processing for fluorescence resonance energy transfer (FRET) measurements; and intracellular spatial distribution measurements of RhoA GTPase activity. Results: The interdivision time was 19.4 h (mean) ± 6.0 h (SD) (n = 7) for the GFP-histone cells in the two-by-two field that was scanned for 72 h. After registration and ratioing of the cells with the RhoA biosensor, increases in both cell protrusion and retraction were coincident with to increases in RhoA activity. Conclusions: These advances lay the foundation for extracting and correlating measurements characterizing the functional relationships of spatial localization and protein activation with features of cell migration such as velocity, polarization, protrusion, retraction, and mitosis.

Original languageEnglish (US)
Pages (from-to)563-572
Number of pages10
JournalCytometry Part A
Volume69
Issue number7
DOIs
StatePublished - Jul 1 2006
Externally publishedYes

Keywords

  • Cell migration
  • Fluorescence
  • High-throughput microscopy
  • Registration alignment
  • Slide-based cytometry

ASJC Scopus subject areas

  • Pathology and Forensic Medicine
  • Histology
  • Cell Biology

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    Shen, F., Hodgson, L., Rabinovich, A., Pertz, O., Hahn, K., & Price, J. H. (2006). Functional proteometrics for cell migration. Cytometry Part A, 69(7), 563-572. https://doi.org/10.1002/cyto.a.20283