Fingerprint-to-CH stretch continuously tunable high spectral resolution stimulated Raman scattering microscope.
Sergey P LaptenokVijayakumar P RajamanickamLuca GenchiTual MonfortYeonwoo LeeImran I PatelAndrea BertonciniCarlo LiberalePublished in: Journal of biophotonics (2019)
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.
Keyphrases
- label free
- optical coherence tomography
- high resolution
- single molecule
- high speed
- raman spectroscopy
- dual energy
- energy transfer
- density functional theory
- mass spectrometry
- computed tomography
- molecular dynamics simulations
- room temperature
- magnetic resonance imaging
- cell death
- deep learning
- dna methylation
- photodynamic therapy
- genome wide
- quality control
- single cell
- risk assessment