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Updated: Jun 13, 2026
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Nanotopology of Cell Adhesion upon Variable-Angle Total Internal Reflection Fluorescence Microscopy (VA-TIRFM)
Published on: October 2, 2012
Live cell adhesion assay with attenuated total reflection infrared spectroscopy.
Martin Schmidt1, Tobias Wolfram, Monika Rumpler
1Institute for Molecular Biophysics, Bar Harbor, Maine 04609, USA. schmidtm@bessy.de
Biointerphases
|April 23, 2010
Summary
Attenuated total reflection (ATR) infrared (IR) spectroscopy dynamically monitors rainbow trout cells. This technique reveals changes in cell morphology and coverage during adhesion and biochemical responses, proving useful for live cell adhesion assays.
Area of Science:
- Biophysics
- Cell Biology
- Spectroscopy
Background:
- Live cell analysis requires non-invasive techniques.
- Understanding cell adhesion is crucial in biological research.
- Attenuated total reflection (ATR) infrared (IR) spectroscopy offers label-free molecular information.
Purpose of the Study:
- To investigate the applicability of ATR IR spectroscopy for real-time monitoring of live fish fibroblast cells.
- To analyze dynamic changes in cell morphology and surface coverage during adhesion and response to biochemical stimuli.
Main Methods:
- Utilized rainbow trout fibroblast cells (RTG-P1) cultured on a diamond surface.
- Employed attenuated total reflection (ATR) infrared (IR) spectroscopy for dynamic spectral recording.
- Applied biochemical agents (trypsin and cytochalasin D) to induce cellular responses.
Main Results:
- Observed distinct changes in IR spectra correlated with cell adsorption and spreading.
- Demonstrated that spectral variations reflect alterations in cell morphology and surface coverage.
- Successfully monitored dynamic cellular responses to trypsin and cytochalasin D in real-time.
Conclusions:
- ATR IR spectroscopy is a viable technique for label-free, real-time monitoring of live cell adhesion.
- The method provides insights into cell morphology and surface interactions.
- This approach holds significant potential for live cell adhesion assays in various biological contexts.
Related Concept Videos
01:13Attenuated Total Reflectance (ATR) Infrared Spectroscopy: Overview
Attenuated total reflectance (ATR) infrared spectroscopy is a powerful analytical technique used to study the composition of materials. It is widely employed in chemistry, materials science, forensic science, and other fields where sample characterization is required. ATR has several advantages over traditional transmission IR spectroscopy, including the requirement of little to no sample preparation and the ability to analyze a wide range of samples.
The ATR process begins by directing a beam...
The ATR process begins by directing a beam...
01:05Total Internal Reflection Fluorescence Microscopy
Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.