Method for High-Resolution Nanoscopy of Whole Cells and Tissues

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Researchers at Purdue University have developed an imaging method for enhanced resolution of 3D biological structures. Single-molecule localization microscopy is a powerful tool in visualizing organelle structures, interactions, and protein functions in biological research. However, whole-cell and tissue specimens challenge the achievable resolution and depth of nanoscopy methods. As imaging depth increases, photons emitted by fluorescent probes are scattered and aberrated, resulting in image artifacts and rapidly deteriorating resolution. The Purdue researchers propose a method of point spread function retrieval for three-dimensional nanoscopy. The system involves multiple components, including a 3D nanoscopy imaging device, an encoder that uses the images to provide point spread functions (PSFs), and a processor that receives PSFs and provides 3D locations of the molecules in a system. The processor uses an in situ PSF retrieval algorithm (INSPR) that iteratively forms 3D PSF stacks for single molecule patterns until the detected and retrieved PSFs match. This system provides limit-achieving precision and uncompromised fidelity through whole cells and tissues.

Related Publications: Three-Dimensional Single-Molecule Localization Microscopy in Whole-Cell and Tissue Specimens. Sheng Liu, Hyun Huh, Sang-Hyuk Lee, Fang Huang. Annual Review of Biomedical Engineering 2020 22:1, 155-184.

Technology Validation: In a test with immuno-labeled TOM20 proteins, the INSPR method successfully imaged the surface contour of each mitochondrial protein complex with high resolution. Compared to an in vitro method, INSPR results had 8 nm higher precision in the lateral direction and 21 nm higher precision in the axial direction.

- Allows imaging of whole cells and tissues
- Allows imaging far from coverslip

- Imaging of 3D biological structures
Jan 18, 2022
United States

Aug 6, 2020
PCT-Gov. Funding

Aug 6, 2019
United States
Purdue Office of Technology Commercialization
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