학술논문
Mesoscopic oblique plane microscopy with a diffractive light-sheet for large-scale 4D cellular resolution imaging.
Document Type
Academic Journal
Author
Shao W; Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21231, USA.; Department of Ophthalmology, Johns Hopkins University, Baltimore, Maryland 21231, USA.; Chang M; Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21231, USA.; Emmerich K; Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21231, USA.; Kanold PO; Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21231, USA.; Mumm JS; Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21231, USA.; Yi J; Department of Biomedical Engineering, Johns Hopkins University, Baltimore, Maryland 21231, USA.; Department of Ophthalmology, Johns Hopkins University, Baltimore, Maryland 21231, USA.
Source
Publisher: Optica Publishing Group Country of Publication: United States NLM ID: 101643595 Publication Model: Print-Electronic Cited Medium: Print ISSN: 2334-2536 (Print) Linking ISSN: 23342536 NLM ISO Abbreviation: Optica Subsets: PubMed not MEDLINE
Subject
Language
English
ISSN
2334-2536
Abstract
Fundamental understanding of large-scale dynamic connectivity within a living organism requires volumetric imaging over a large field of view (FOV) at biologically relevant speed and resolution. However, most microscopy methods make trade-offs between FOV and axial resolution, making it challenging to observe highly dynamic processes at cellular resolution in 3D across mesoscopic scales (e.g., whole zebrafish larva). To overcome this limitation, we have developed mesoscopic oblique plane microscopy (Meso-OPM) with a diffractive light sheet. By augmenting the illumination angle of the light sheet with a transmission grating, we improved the axial resolution approximately sixfold over existing methods and approximately twofold beyond the diffraction limitation of the primary objective lens. We demonstrated a FOV up to 5.4 mm × 3.3 mm with resolution of 2.5 μm × 3 μm × 6 μm, allowing volumetric imaging of 3D cellular structures with a single scan. Applying Meso-OPM for in vivo imaging of zebrafish larvae, we report here in toto whole-body volumetric recordings of neuronal activity at 2 Hz volume rate and whole-body volumetric recordings of blood flow dynamics at 5 Hz with 3D cellular resolution.
Competing Interests: Disclosures. All other authors declare they have no competing interests. We acknowledge the Ahrens Lab of the Howard Hughes Medical Institute’s Janelia Research Campus for providing the jGCaMP7s expressing Zebrafish larvae.
Competing Interests: Disclosures. All other authors declare they have no competing interests. We acknowledge the Ahrens Lab of the Howard Hughes Medical Institute’s Janelia Research Campus for providing the jGCaMP7s expressing Zebrafish larvae.