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Updated: Jul 9, 2026
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Simultaneous Brightfield, Fluorescence, and Optical Coherence Tomographic Imaging of Contracting Cardiac Trabeculae Ex Vivo
Published on: October 2, 2021
Combined scanning optical coherence and two-photon-excited fluorescence microscopy
Optics Letters
|December 13, 2007
Summary
We developed a novel method for simultaneous optical coherence microscopy (OCM) and two-photon-excited (TPE) fluorescence imaging. This technique achieves rapid, submicrometer resolution imaging of biological samples, including live embryos.
Area of Science:
- Biomedical Optics
- Microscopy
- Cell Biology
Background:
- Simultaneous imaging modalities offer enhanced biological insight.
- Optical coherence microscopy (OCM) provides structural information.
- Two-photon-excited (TPE) fluorescence microscopy visualizes molecular and cellular functions.
Purpose of the Study:
- To demonstrate simultaneous imaging using OCM and TPE fluorescence microscopy.
- To achieve high-resolution, rapid en-face imaging.
- To validate the technique in live biological samples.
Main Methods:
- A mode-locked Ti:sapphire laser was used for simultaneous OCM and TPE excitation.
- The laser was focused and scanned in three dimensions through a fixed sample.
- Backscattered light (OCM) and fluorescence light (TPE) were independently detected.
Main Results:
- Both OCM and TPE modes achieved rapid en-face imaging with submicrometer resolution.
- OCM demonstrated high sensitivity (>130 dB) at 100-kHz pixel rates.
- Simultaneous imaging of cell nuclei in live Drosophila embryos was successfully achieved.
Conclusions:
- Simultaneous OCM and TPE microscopy is a viable technique for high-resolution biological imaging.
- This dual-modality approach provides complementary structural and functional information.
- The method is suitable for studying dynamic processes in live biological systems.
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