Broadband stimulated Raman imaging based on multi-channel lock-in detection for spectral histopathology
Spontaneous Raman microscopy reveals the chemical composition of a sample in a label-free and non-invasive fashion by directly measuring the vibrational spectra of molecules. However, its extremely low cross section prevents its application to fast imaging. Stimulated Raman scattering (SRS) amplifie...
| Main Authors: | , , , , , , , , , , , , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
AIP Publishing LLC
2022-07-01
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| Series: | APL Photonics |
| Online Access: | http://dx.doi.org/10.1063/5.0093946 |
| _version_ | 1828151716297572352 |
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| author | Alejandro De la Cadena Federico Vernuccio Andrea Ragni Giuseppe Sciortino Renzo Vanna Carino Ferrante Natalia Pediconi Carlo Valensise Luca Genchi Sergey P. Laptenok Andrea Doni Marco Erreni Tullio Scopigno Carlo Liberale Giorgio Ferrari Marco Sampietro Giulio Cerullo Dario Polli |
| author_facet | Alejandro De la Cadena Federico Vernuccio Andrea Ragni Giuseppe Sciortino Renzo Vanna Carino Ferrante Natalia Pediconi Carlo Valensise Luca Genchi Sergey P. Laptenok Andrea Doni Marco Erreni Tullio Scopigno Carlo Liberale Giorgio Ferrari Marco Sampietro Giulio Cerullo Dario Polli |
| author_sort | Alejandro De la Cadena |
| collection | DOAJ |
| description | Spontaneous Raman microscopy reveals the chemical composition of a sample in a label-free and non-invasive fashion by directly measuring the vibrational spectra of molecules. However, its extremely low cross section prevents its application to fast imaging. Stimulated Raman scattering (SRS) amplifies the signal by several orders of magnitude thanks to the coherent nature of the nonlinear process, thus unlocking high-speed microscopy applications that provide analytical information to elucidate biochemical mechanisms with subcellular resolution. Nevertheless, in its standard implementation, narrowband SRS provides images at only one frequency at a time, which is not sufficient to distinguish constituents with overlapping Raman bands. Here, we report a broadband SRS microscope equipped with a home-built multichannel lock-in amplifier simultaneously measuring the SRS signal at 32 frequencies with integration time down to 44 µs, allowing for detailed, high spatial resolution mapping of spectrally congested samples. We demonstrate the capability of our microscope to differentiate the chemical constituents of heterogeneous samples by measuring the relative concentrations of different fatty acids in cultured hepatocytes at the single lipid droplet level and by differentiating tumor from peritumoral tissue in a preclinical mouse model of fibrosarcoma. |
| first_indexed | 2024-04-11T22:00:37Z |
| format | Article |
| id | doaj.art-b717adcfe216410fb66c53f294e9b26d |
| institution | Directory Open Access Journal |
| issn | 2378-0967 |
| language | English |
| last_indexed | 2024-04-11T22:00:37Z |
| publishDate | 2022-07-01 |
| publisher | AIP Publishing LLC |
| record_format | Article |
| series | APL Photonics |
| spelling | doaj.art-b717adcfe216410fb66c53f294e9b26d2022-12-22T04:00:56ZengAIP Publishing LLCAPL Photonics2378-09672022-07-0177076104076104-1210.1063/5.0093946Broadband stimulated Raman imaging based on multi-channel lock-in detection for spectral histopathologyAlejandro De la Cadena0Federico Vernuccio1Andrea Ragni2Giuseppe Sciortino3Renzo Vanna4Carino Ferrante5Natalia Pediconi6Carlo Valensise7Luca Genchi8Sergey P. Laptenok9Andrea Doni10Marco Erreni11Tullio Scopigno12Carlo Liberale13Giorgio Ferrari14Marco Sampietro15Giulio Cerullo16Dario Polli17Physics Department, Politecnico di Milano, Milan, ItalyPhysics Department, Politecnico di Milano, Milan, ItalyElectronics, Information, and Bioengineering Department, Politecnico di Milano, ItalyElectronics, Information, and Bioengineering Department, Politecnico di Milano, ItalyInstitute for Photonics and Nanotechnologies, CNR (IFN-CNR), Milan, ItalyPhysics Department, Universitá di Roma “La Sapienza,” Roma, ItalyPhysics Department, Universitá di Roma “La Sapienza,” Roma, ItalyEnrico Fermi Research Center, Roma, ItalyBiological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology (KAUST), 23955 Thuwal, Saudi ArabiaBiological and Environmental Science and Engineering Division, King Abdullah University of Science and Technology (KAUST), 23955 Thuwal, Saudi ArabiaUnit of Advanced Optical Microscopy, IRCCS Humanitas Research Hospital, Milan, ItalyUnit of Advanced Optical Microscopy, IRCCS Humanitas Research Hospital, Milan, ItalyPhysics Department, Universitá di Roma “La Sapienza,” Roma, ItalyComputer, Electrical and Mathematical Sciences Division, King Abdullah University of Science and Technology (KAUST), 23955 Thuwal, Saudi ArabiaElectronics, Information, and Bioengineering Department, Politecnico di Milano, ItalyElectronics, Information, and Bioengineering Department, Politecnico di Milano, ItalyPhysics Department, Politecnico di Milano, Milan, ItalyPhysics Department, Politecnico di Milano, Milan, ItalySpontaneous Raman microscopy reveals the chemical composition of a sample in a label-free and non-invasive fashion by directly measuring the vibrational spectra of molecules. However, its extremely low cross section prevents its application to fast imaging. Stimulated Raman scattering (SRS) amplifies the signal by several orders of magnitude thanks to the coherent nature of the nonlinear process, thus unlocking high-speed microscopy applications that provide analytical information to elucidate biochemical mechanisms with subcellular resolution. Nevertheless, in its standard implementation, narrowband SRS provides images at only one frequency at a time, which is not sufficient to distinguish constituents with overlapping Raman bands. Here, we report a broadband SRS microscope equipped with a home-built multichannel lock-in amplifier simultaneously measuring the SRS signal at 32 frequencies with integration time down to 44 µs, allowing for detailed, high spatial resolution mapping of spectrally congested samples. We demonstrate the capability of our microscope to differentiate the chemical constituents of heterogeneous samples by measuring the relative concentrations of different fatty acids in cultured hepatocytes at the single lipid droplet level and by differentiating tumor from peritumoral tissue in a preclinical mouse model of fibrosarcoma.http://dx.doi.org/10.1063/5.0093946 |
| spellingShingle | Alejandro De la Cadena Federico Vernuccio Andrea Ragni Giuseppe Sciortino Renzo Vanna Carino Ferrante Natalia Pediconi Carlo Valensise Luca Genchi Sergey P. Laptenok Andrea Doni Marco Erreni Tullio Scopigno Carlo Liberale Giorgio Ferrari Marco Sampietro Giulio Cerullo Dario Polli Broadband stimulated Raman imaging based on multi-channel lock-in detection for spectral histopathology APL Photonics |
| title | Broadband stimulated Raman imaging based on multi-channel lock-in detection for spectral histopathology |
| title_full | Broadband stimulated Raman imaging based on multi-channel lock-in detection for spectral histopathology |
| title_fullStr | Broadband stimulated Raman imaging based on multi-channel lock-in detection for spectral histopathology |
| title_full_unstemmed | Broadband stimulated Raman imaging based on multi-channel lock-in detection for spectral histopathology |
| title_short | Broadband stimulated Raman imaging based on multi-channel lock-in detection for spectral histopathology |
| title_sort | broadband stimulated raman imaging based on multi channel lock in detection for spectral histopathology |
| url | http://dx.doi.org/10.1063/5.0093946 |
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