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Fingerprint‐to‐CH stretch continuously tunable high spectral resolution stimulated Raman scattering microscope
Author(s) -
Laptenok Sergey P.,
Rajamanickam Vijayakumar P.,
Genchi Luca,
Monfort Tual,
Lee Yeonwoo,
Patel Imran I.,
Bertoncini Andrea,
Liberale Carlo
Publication year - 2019
Publication title -
journal of biophotonics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.877
H-Index - 66
eISSN - 1864-0648
pISSN - 1864-063X
DOI - 10.1002/jbio.201900028
Subject(s) - chemical imaging , hyperspectral imaging , raman spectroscopy , microscope , microscopy , spectral resolution , raman scattering , materials science , spectral imaging , resolution (logic) , spectral signature , raman microscope , optics , optical microscope , fingerprint (computing) , imaging spectroscopy , image resolution , scanning electron microscope , spectral line , remote sensing , computer science , physics , artificial intelligence , astronomy , geology
Stimulated Raman scattering (SRS) microscopy is a label‐free method generating images based on chemical contrast within samples, and has already shown its great potential for high‐sensitivity and fast imaging of biological specimens. The capability of SRS to collect molecular vibrational signatures in bio‐samples, coupled with the availability of powerful statistical analysis methods, allows quantitative chemical imaging of live cells with sub‐cellular resolution. This application has substantially driven the development of new SRS microscopy platforms. Indeed, in recent years, there has been a constant effort on devising configurations able to rapidly collect Raman spectra from samples over a wide vibrational spectral range, as needed for quantitative analysis by using chemometric methods. In this paper, an SRS microscope which exploits spectral shaping by a narrowband and rapidly tunable acousto‐optical tunable filter (AOTF) is presented. This microscope enables spectral scanning from the Raman fingerprint region to the Carbon‐Hydrogen (CH)‐stretch region without any modification of the optical setup. Moreover, it features also a high enough spectral resolution to allow resolving Raman peaks in the crowded fingerprint region. Finally, application of the developed SRS microscope to broadband hyperspectral imaging of biological samples over a large spectral range from 800 to 3600 cm −1 , is demonstrated.