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Multimodal Nonlinear Hyperspectral Chemical Imaging Using Line-Scanning Vibrational Sum-Frequency Generation Microscopy
Published on: December 1, 2023
Microscope enabling multimodality imaging, angle-resolved scattering, and scattering spectroscopy
W J Cottrell1, J D Wilson, T H Foster
1Institute of Optics and Departments of Physics and of Imaging Sciences, University of Rochester, Rochester, NY 14642, USA.
Optics Letters
|August 19, 2007
Summary
We developed a versatile microscope system for simultaneous imaging and light scattering spectroscopy. This advanced tool aids in studying subcellular biophysics and cell behavior using light scattering and fluorescence techniques.
Area of Science:
- Biophysics
- Optical Microscopy
- Spectroscopy
Background:
- Multimodal optical microscopy is crucial for understanding cellular processes.
- Integrating diverse imaging and spectroscopy techniques into a single platform presents significant technical challenges.
- Characterizing subcellular structures requires high-resolution imaging and sensitive spectroscopic analysis.
Purpose of the Study:
- To design and build a multifunctional imaging and scattering spectroscopy system.
- To enable co-registered acquisition of various optical signals from the same field of view.
- To provide a platform for investigating the biophysical implications of subcellular light scattering in conjunction with fluorescence microscopy.
Main Methods:
- A commercial inverted microscope was adapted with a custom fiber-based illumination system.
- The system integrates brightfield, Fourier-filtered darkfield, and fluorescence imaging.
- Monochromatic angle-resolved scattering and white-light wavelength-resolved scattering spectroscopy were implemented.
Main Results:
- The system successfully integrates multiple imaging and spectroscopy modalities.
- Experimental verification using fluorescent microspheres confirmed the performance of each data acquisition mode.
- Co-registered data acquisition from the same field of view was achieved.
Conclusions:
- The developed multifunctional instrument offers a powerful platform for advanced cell biology research.
- It enables simultaneous investigation of subcellular light scattering and fluorescence.
- This integrated approach facilitates studies on the biophysical properties of adherent cells.
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