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Cortical Actin Flow in T Cells Quantified by Spatio-temporal Image Correlation Spectroscopy of Structured Illumination Microscopy Data
Published on: December 17, 2015
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Epi-illumination SPIM for volumetric imaging with high spatial-temporal resolution.
Bin Yang1, Xingye Chen1,2, Yina Wang1
1Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA, USA.
Nature Methods
|May 8, 2019
Summary
We developed a new light-sheet microscopy system for faster, higher-resolution imaging. This advanced microscopy technique achieves single-molecule sensitivity and is compatible with standard lab equipment, enabling detailed cellular studies.
Area of Science:
- Biophysics
- Microscopy
- Cell Biology
Background:
- Traditional microscopy techniques face limitations in speed, resolution, and sample compatibility for advanced cellular studies.
- There is a need for microscopy systems that offer high resolution and sensitivity while integrating seamlessly with existing laboratory workflows.
Purpose of the Study:
- To design and validate a novel epi-illumination single-plane illumination microscopy (SPIM) system.
- To achieve subcellular resolution and single-molecule sensitivity with a simplified optical setup.
- To demonstrate the system's compatibility with standard biological sample holders and its capability for advanced imaging applications.
Main Methods:
- Development of an epi-illumination SPIM system utilizing a single objective lens.
- Integration of a sample interface identical to inverted fluorescence microscopes, avoiding additional reflection elements.
- Implementation of multicolor fast volumetric imaging, single-molecule localization microscopy (SMLM), and parallel imaging capabilities.
Main Results:
- The developed SPIM system achieved subcellular resolution and single-molecule sensitivity.
- Demonstrated multicolor fast volumetric imaging and single-molecule localization microscopy.
- Successfully performed parallel imaging of 16 cell lines and recorded cellular responses to perturbations.
Conclusions:
- The novel epi-illumination SPIM system offers a versatile and powerful platform for advanced biological imaging.
- Its compatibility with common sample holders and simplified design enhance its applicability in diverse research settings.
- The system enables high-throughput and high-resolution cellular analysis, advancing cell biology and biophysics research.
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