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Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix

Live-cell Imaging of Migrating Cells Expressing Fluorescently-tagged Proteins in a Three-dimensional Matrix

Published on: December 22, 2011

High resolution multimode light microscopy of cell migration: long-term imaging and analysis.

Torsten Wöllert¹, George M Langford

¹Department of Biology, Life Sciences Complex, Syracuse University, Syracuse, NY, USA.

Methods in Molecular Biology (Clifton, N.J.)

|September 22, 2009

Summary

This study explores cell migration using human oral keratinocytes (OKF6/TERT-2) to understand complex regulation. Advanced microscopy techniques enable long-term observation of dynamic cellular processes during infection.

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Non-invasive Imaging of Leukocyte Homing and Migration in vivo

Non-invasive Imaging of Leukocyte Homing and Migration in vivo

Published on: December 5, 2010

Area of Science:

Cell Biology
Microscopy
Immunology

Background:

Cell migration is crucial for development, healing, and disease, involving cytoskeleton and extracellular matrix dynamics.
Keratinocytes migrate in sheets, responding to stimuli like pathogens, requiring long-term observation for regulatory insights.

Purpose of the Study:

To illustrate complex cell migration patterns and regulation using human oral keratinocytes (OKF6/TERT-2).
To analyze spatio-temporal regulation of the cytoskeleton and signaling pathways during infection using long-term live cell imaging.

Main Methods:

Utilized human oral keratinocytes (OKF6/TERT-2) for studying cell migration.
Employed two multimode digital imaging systems (VEC-DIC and BioStation IM) for long-term, high-resolution time-lapse imaging.
Incorporated wide-field epifluorescence and transmitted light microscopy for detailed motion analysis.

Main Results:

Demonstrated complex migration patterns in keratinocyte sheets responding to Candida albicans.
Observed dynamic changes in cytoskeleton and cell-cell/substrate interactions over extended periods.
Validated the utility of advanced imaging systems for capturing spatio-temporal cellular dynamics.

Conclusions:

Long-term live cell imaging is essential for understanding the complex regulation of cell migration, especially during infection.
The described imaging systems provide high-resolution, stable environments for observing dynamic cellular processes.
This research provides a foundation for further studies into cell migration mechanisms and therapeutic targets.