Full-Spectrum CARS Microscopy of Cells and Tissues with Ultrashort White-Light Continuum Pulses
Coherent anti-Stokes Raman scattering (CARS) microscopy is an emerging nonlinear vibrational imaging technique that delivers label-free chemical maps of cells and tissues. In narrowband CARS, two spatiotemporally superimposed picosecond pulses, pump and Stokes, illuminate the sample to interrogate a...
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| Veröffentlicht in: | The journal of physical chemistry. B 2023-06, Vol.127 (21), p.4733-4745 |
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| creator | Vernuccio, Federico Vanna, Renzo Ceconello, Chiara Bresci, Arianna Manetti, Francesco Sorrentino, Salvatore Ghislanzoni, Silvia Lambertucci, Flavia Motiño, Omar Martins, Isabelle Kroemer, Guido Bongarzone, Italia Cerullo, Giulio Polli, Dario |
| description | Coherent anti-Stokes Raman scattering (CARS) microscopy is an emerging nonlinear vibrational imaging technique that delivers label-free chemical maps of cells and tissues. In narrowband CARS, two spatiotemporally superimposed picosecond pulses, pump and Stokes, illuminate the sample to interrogate a single vibrational mode. Broadband CARS (BCARS) combines narrowband pump pulses with broadband Stokes pulses to record broad vibrational spectra. Despite recent technological advancements, BCARS microscopes still struggle to image biological samples over the entire Raman-active region (400–3100 cm–1). Here, we demonstrate a robust BCARS platform that answers this need. Our system is based on a femtosecond ytterbium laser at a 1035 nm wavelength and a 2 MHz repetition rate, which delivers high-energy pulses used to produce broadband Stokes pulses by white-light continuum generation in a bulk YAG crystal. Combining such pulses, pre-compressed to sub-20 fs duration, with narrowband pump pulses, we generate a CARS signal with a high ( |
| doi_str_mv | 10.1021/acs.jpcb.3c01443 |
| format | Article |
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In narrowband CARS, two spatiotemporally superimposed picosecond pulses, pump and Stokes, illuminate the sample to interrogate a single vibrational mode. Broadband CARS (BCARS) combines narrowband pump pulses with broadband Stokes pulses to record broad vibrational spectra. Despite recent technological advancements, BCARS microscopes still struggle to image biological samples over the entire Raman-active region (400–3100 cm–1). Here, we demonstrate a robust BCARS platform that answers this need. Our system is based on a femtosecond ytterbium laser at a 1035 nm wavelength and a 2 MHz repetition rate, which delivers high-energy pulses used to produce broadband Stokes pulses by white-light continuum generation in a bulk YAG crystal. Combining such pulses, pre-compressed to sub-20 fs duration, with narrowband pump pulses, we generate a CARS signal with a high (<9 cm–1) spectral resolution in the whole Raman-active window, exploiting both the two-color and three-color excitation mechanisms. Aided by an innovative post-processing pipeline, our microscope allows us to perform high-speed (≈1 ms pixel dwell time) imaging over a large field of view, identifying the main chemical compounds in cancer cells and discriminating tumorous from healthy regions in liver slices of mouse models, paving the way for applications in histopathological settings.</description><identifier>ISSN: 1520-6106</identifier><identifier>EISSN: 1520-5207</identifier><identifier>DOI: 10.1021/acs.jpcb.3c01443</identifier><identifier>PMID: 37195090</identifier><language>eng</language><publisher>United States: American Chemical Society</publisher><subject>Animals ; B: Biophysical and Biochemical Systems and Processes ; Bioengineering ; Biological Physics ; Human health and pathology ; Imaging ; Lasers ; Life Sciences ; Light ; Mice ; Microscopy ; Nonlinear Optical Microscopy ; Physics ; Spectrum Analysis, Raman - methods ; Tissues and Organs</subject><ispartof>The journal of physical chemistry. 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B</addtitle><date>2023-06-01</date><risdate>2023</risdate><volume>127</volume><issue>21</issue><spage>4733</spage><epage>4745</epage><pages>4733-4745</pages><issn>1520-6106</issn><eissn>1520-5207</eissn><abstract>Coherent anti-Stokes Raman scattering (CARS) microscopy is an emerging nonlinear vibrational imaging technique that delivers label-free chemical maps of cells and tissues. In narrowband CARS, two spatiotemporally superimposed picosecond pulses, pump and Stokes, illuminate the sample to interrogate a single vibrational mode. Broadband CARS (BCARS) combines narrowband pump pulses with broadband Stokes pulses to record broad vibrational spectra. Despite recent technological advancements, BCARS microscopes still struggle to image biological samples over the entire Raman-active region (400–3100 cm–1). Here, we demonstrate a robust BCARS platform that answers this need. Our system is based on a femtosecond ytterbium laser at a 1035 nm wavelength and a 2 MHz repetition rate, which delivers high-energy pulses used to produce broadband Stokes pulses by white-light continuum generation in a bulk YAG crystal. Combining such pulses, pre-compressed to sub-20 fs duration, with narrowband pump pulses, we generate a CARS signal with a high (<9 cm–1) spectral resolution in the whole Raman-active window, exploiting both the two-color and three-color excitation mechanisms. Aided by an innovative post-processing pipeline, our microscope allows us to perform high-speed (≈1 ms pixel dwell time) imaging over a large field of view, identifying the main chemical compounds in cancer cells and discriminating tumorous from healthy regions in liver slices of mouse models, paving the way for applications in histopathological settings.</abstract><cop>United States</cop><pub>American Chemical Society</pub><pmid>37195090</pmid><doi>10.1021/acs.jpcb.3c01443</doi><tpages>13</tpages><orcidid>https://orcid.org/0000-0001-9125-5225</orcidid><orcidid>https://orcid.org/0000-0002-9534-2702</orcidid><orcidid>https://orcid.org/0000-0002-6960-5708</orcidid><orcidid>https://orcid.org/0000-0002-4281-7524</orcidid><orcidid>https://orcid.org/0000-0002-9334-4405</orcidid><oa>free_for_read</oa></addata></record> |
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| subjects | Animals B: Biophysical and Biochemical Systems and Processes Bioengineering Biological Physics Human health and pathology Imaging Lasers Life Sciences Light Mice Microscopy Nonlinear Optical Microscopy Physics Spectrum Analysis, Raman - methods Tissues and Organs |
| title | Full-Spectrum CARS Microscopy of Cells and Tissues with Ultrashort White-Light Continuum Pulses |
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