Integrating SHG and two-color STED microscopy for super-resolved imaging of cell-matrix interactions in myocardium

Zhao Zhang; Adam Baker; Hongming Fan; William Richardson; Thomas K. Borg; Bruce Z. Gao; Tong Ye

doi:10.1117/12.2610374

2 March 2022 Integrating SHG and two-color STED microscopy for super-resolved imaging of cell-matrix interactions in myocardium

Zhao Zhang, Adam Baker, Hongming Fan, William Richardson, Thomas K. Borg, Bruce Z. Gao, Tong Ye

Author Affiliations +

Proceedings Volume 11967, Single Molecule Spectroscopy and Superresolution Imaging XV; 1196708 (2022) https://doi.org/10.1117/12.2610374
Event: SPIE BiOS, 2022, San Francisco, California, United States

Abstract

Understanding cardiomyocyte-extracellular matrix (ECM) interactions at the molecular level is essential for deeper insights into their mechanical signaling function for cardiac development, homeostasis and remodeling. We report a lab-built microscope integrating two-color STED microscopy with second harmonic generation (SHG) microscopy to investigate the detailed architecture of cardiomyocyte-ECM interactions in murine myocardium at a subdiffractive level. SHG microscopy is used to locate possible interaction sites at the cell-ECM interface through the intrinsic SHG signal generated by collagen assemblies and myosin filaments. Two-color STED microscopy is used to obtain a subdiffractive view of proteins at sites of interest registered by SHG microscopy. Because large field-of-view (FOV) STED microscopy is still challenging, with photobleaching often a major concern, imaging only SHG-registered sites is advantageous. Further, using intrinsic contrast in the study reduces the number of biomarkers for fluorescent staining and thereby the number of detection channels for fluorescent imaging, simplifying sample preparation procedures and STED microscopy architectures. For purpose of demonstration, we show images of immunostained type I collagen, type Ⅳ collagen and laminin as ECM structures of interest in rat ventricular sections without counterstaining.

Conference Presentation

Citation Download Citation

Zhao Zhang, Adam Baker, Hongming Fan, William Richardson, Thomas K. Borg, Bruce Z. Gao, and Tong Ye "Integrating SHG and two-color STED microscopy for super-resolved imaging of cell-matrix interactions in myocardium", Proc. SPIE 11967, Single Molecule Spectroscopy and Superresolution Imaging XV, 1196708 (2 March 2022); https://doi.org/10.1117/12.2610374

ACCESS THE FULL ARTICLE

INSTITUTIONAL
Select your institution to access the SPIE Digital Library.

SELECT YOUR INSTITUTION

PERSONAL
Sign in with your SPIE account to access your personal subscriptions or to use specific features such as save to my library, sign up for alerts, save searches, etc.

PERSONAL SIGN IN

No SPIE Account? Create one

PURCHASE THIS CONTENT

SUBSCRIBE TO DIGITAL LIBRARY

50 downloads per 1-year subscription

Members: $195

Non-members: $335 ADD TO CART

25 downloads per 1 - year subscription

Members: $145

Non-members: $250 ADD TO CART

PURCHASE SINGLE ARTICLE

Includes PDF, HTML & Video, when available