Significance: The reprojection setup typical of oblique plane microscopy (OPM) limits the effective aperture of the imaging system, and therefore its efficiency and resolution. Large aperture system is only possible through the use of custom specialized optics. A full-aperture OPM made with off the shelf components would both improve the performance of the method and encourage its widespread adoption. Aim: To prove the feasibility of an OPM without a conventional reprojection setup, retaining the full aperture of the primary objective employed. Approach: A deformable lens based remote focusing setup synchronized with the rolling shutter of a complementary metal-oxide semiconductor detector is used instead of a traditional reprojection system. Results: The system was tested on microbeads, prepared slides, and zebrafish embryos. Resolution and pixel throughput were superior to conventional OPM with cropped apertures, and comparable with OPM implementations with custom made optical components. Conclusions: An easily reproducible approach to OPM imaging is presented, eliminating the conventional reprojection setup and exploiting the full aperture of the employed objective.

Full-aperture extended-depth oblique plane microscopy through dynamic remote focusing / P. Pozzi, V. Balan, A. Candeo, A. Brix, A.S. Pistocchi, C. D'Andrea, G. Valentini, A. Bassi. - In: JOURNAL OF BIOMEDICAL OPTICS. - ISSN 1560-2281. - 29:3(2024 Mar 29), pp. 036502.1-036502.12. [10.1117/1.JBO.29.3.036502]

Full-aperture extended-depth oblique plane microscopy through dynamic remote focusing

A. Brix;A.S. Pistocchi;
2024

Abstract

Significance: The reprojection setup typical of oblique plane microscopy (OPM) limits the effective aperture of the imaging system, and therefore its efficiency and resolution. Large aperture system is only possible through the use of custom specialized optics. A full-aperture OPM made with off the shelf components would both improve the performance of the method and encourage its widespread adoption. Aim: To prove the feasibility of an OPM without a conventional reprojection setup, retaining the full aperture of the primary objective employed. Approach: A deformable lens based remote focusing setup synchronized with the rolling shutter of a complementary metal-oxide semiconductor detector is used instead of a traditional reprojection system. Results: The system was tested on microbeads, prepared slides, and zebrafish embryos. Resolution and pixel throughput were superior to conventional OPM with cropped apertures, and comparable with OPM implementations with custom made optical components. Conclusions: An easily reproducible approach to OPM imaging is presented, eliminating the conventional reprojection setup and exploiting the full aperture of the employed objective.
deformable lenses; fluorescence; light-sheet; microscopy; oblique plane microscopy; remote focusing;
Settore BIO/13 - Biologia Applicata
   The Integrated Initiative of European Laser Research Infrastructures
   LASERLAB-EUROPE
   European Commission
   Horizon 2020 Framework Programme
   871124
29-mar-2024
21-mar-2024
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/1043552
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