Super-resolution Vibrational Imaging Based on Photoswitchable Raman Probe

Overview

Journal Sci Adv

Specialties Biology
Science

Date 2023 Jun 16

PMID 37327330

Authors

Jingwen Shou

Ayumi Komazawa

Yuusaku Wachi

Minoru Kawatani

Hiroyoshi Fujioka

Spencer John Spratt

Takaha Mizuguchi

Kenichi Oguchi

Hikaru Akaboshi

Fumiaki Obata

Ryo Tachibana

Shun Yasunaga

Yoshio Mita

Yoshihiro Misawa

Ryosuke Kojima

Yasuteru Urano

Mako Kamiya

Yasuyuki Ozeki

Affiliations

Soon will be listed here.

Abstract

Super-resolution vibrational microscopy is promising to increase the degree of multiplexing of nanometer-scale biological imaging because of the narrower spectral linewidth of molecular vibration compared to fluorescence. However, current techniques of super-resolution vibrational microscopy suffer from various limitations including the need for cell fixation, high power loading, or complicated detection schemes. Here, we present reversible saturable optical Raman transitions (RESORT) microscopy, which overcomes these limitations by using photoswitchable stimulated Raman scattering (SRS). We first describe a bright photoswitchable Raman probe (DAE620) and validate its signal activation and depletion characteristics when exposed to low-power (microwatt level) continuous-wave laser light. By harnessing the SRS signal depletion of DAE620 through a donut-shaped beam, we demonstrate super-resolution vibrational imaging of mammalian cells with excellent chemical specificity and spatial resolution beyond the optical diffraction limit. Our results indicate RESORT microscopy to be an effective tool with high potential for multiplexed super-resolution imaging of live cells.

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