Three‐photon‐resonance‐enhanced third‐harmonic generation for label‐free deep‐brain imaging: In search of a chemical contrast
Abstract
Within the past decade, nonlinear Raman microscopy has earned a well‐deserved status of a gold‐standard technology for chemically selective imaging. Even though second‐ and third‐harmonic microscopy is much less demanding on a laser source and multifrequency beam arrangement, it is increasingly falling behind nonlinear Raman scattering as a method of bioimaging because it offers no mechanism whereby imaging could be made chemically specific. Here, we show, however, that such a mechanism does exist, helping harmonic‐generation microscopy overcome its no‐chemical‐specificity handicap. We demonstrate that, with the laser wavelength tuned to a three‐photon resonance with the Soret band of hemoglobin, third‐harmonic generation provides a chemically specific method for a high‐contrast imaging of red blood cells in a broad class of biological systems, including live brain. Moreover, third‐harmonic generation imaging can be conveniently combined with second‐harmonic microscopy on a compact laser platform, providing, as our experiments on rat brain show, a powerful resource for three‐dimensional, cell‐specific label‐free deep‐brain imaging.
Document Details
- Document Type
- Pub Defense Publication
- Publication Date
- Apr 01, 2019
- Source ID
- 10.1002/jrs.5566
Entities
People
- Aleksandr A. Lanin
- Aleksey Zheltikov
- Andrei B. Fedotov
- Artem S. Chebotarev
- Ilya V Kelmanson
- Matvey S. Pochechuev
- Vsevolod V. Belousov
Organizations
- Institute of Bioorganic Chemistry
- Moscow Institute of Physics and Technology
- Moscow State University
- National Research Centre Kurchatov Institute
- Office of Naval Research
- Robert A. Welch Foundation
- Russian Center for Science Information
- Russian Quantum Center
- Russian Science Foundation