Optical sectioning in multi-foci Raman hyperspectral imaging

Liao, Zhiyu; Sinjab, Faris; Elsheikha, Hany M.; Notingher, Ioan

Abstract

In this study, we compared the depth-discrimination and speed performance of multi-foci Raman hyperspectral imaging with the reference standard of a single laser point confocal Raman mapping. A liquid crystal spatial light modulator (LC-SLM) was employed for the generation of multi-foci laser beams, and a digital micromirror device (DMD) was used as a software-configurable reflective pinhole array. The patterns of the laser-foci and pinhole array can be rapidly changed without requiring any hardware alterations. Confocal patterns with different distance-to-size ratios were tested and compared. After optimisation of the laser foci pattern, we demonstrated the feasibility of multi-foci Raman hyperspectral microscopy for recording depth-resolved molecular maps of biological cells (Acanthamoeba castellanii trophozoites). Micrometric depth-discrimination and short acquisition times (20 minutes for single plane confocal image) was achieved.

Citation

Liao, Z., Sinjab, F., Elsheikha, H. M., & Notingher, I. (2018). Optical sectioning in multi-foci Raman hyperspectral imaging. Journal of Raman Spectroscopy, 49(10), 1660-1667. https://doi.org/10.1002/jrs.5450

• Journal Article Type: Article
• Acceptance Date: Jun 20, 2018
• Online Publication Date: Jul 18, 2018
• Publication Date: 2018-10
• Deposit Date: Jul 5, 2018
• Publicly Available Date: Jul 18, 2018
• Journal: Journal of Raman Spectroscopy
• Print ISSN: 0377-0486
• Electronic ISSN: 1097-4555
• Publisher: Wiley
• Peer Reviewed: Peer Reviewed
• Volume: 49
• Issue: 10
• Pages: 1660-1667
• DOI: https://doi.org/10.1002/jrs.5450
• Keywords: Confocal Raman imaging; optical sectioning; multi-beam; cross-talk; single cell imaging
• Public URL: https://nottingham-repository.worktribe.com/output/947176
• Publisher URL: https://onlinelibrary.wiley.com/doi/full/10.1002/jrs.5450
• Contract Date: Jul 5, 2018