Abstract

.Significance: Coherent Raman scattering (CRS) microscopy is an optical imaging technique with capabilities that could benefit a broad range of biomedical research studies.Aim: We reflect on the birth, rapid rise, and inescapable growing pains of the technique and look back on nearly four decades of developments to examine where the CRS imaging approach might be headed in the next decade to come.Approach: We provide a brief historical account of CRS microscopy, followed by a discussion of the challenges to disseminate the technique to a larger audience. We then highlight recent progress in expanding the capabilities of the CRS microscope and assess its current appeal as a practical imaging tool.Results: New developments in Raman tagging have improved the specificity and sensitivity of the CRS technique. In addition, technical advances have led to CRS microscopes that can capture hyperspectral data cubes at practical acquisition times. These improvements have broadened the application space of the technique.Conclusion: The technical performance of the CRS microscope has improved dramatically since its inception, but these advances have not yet translated into a substantial user base beyond a strong core of enthusiasts. Nonetheless, new developments are poised to move the unique capabilities of the technique into the hands of more users.

Highlights

  • New developments are poised to move the unique capabilities of the technique into the hands of more users

  • It was recognized that the Coherent Raman scattering (CRS) signal levels could “exceed those produced in spontaneous Raman processes”[3] and that spurious fluorescence contributions could be largely avoided, an application space in which the attributes of CRS microscopy made a clear difference stayed out of sight

  • Even after substantial improvements to the CRS microscope were reported in 1999,4 experts in the field of optical microscopy continued to be skeptical about the prospects of the stillnascent CRS technology.[5]

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Summary

Results

New developments in Raman tagging have improved the specificity and sensitivity of the CRS technique. Technical advances have led to CRS microscopes that can capture hyperspectral data cubes at practical acquisition times. These improvements have broadened the application space of the technique

Conclusion
New Imaging Technique is Born
Growing Pains and a Renewed Focus
Specificity
Sensitivity
Taking Stock and Looking Ahead
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