PUBLICATION
Real-time volumetric microscopy of in vivo dynamics and large-scale samples with SCAPE 2.0
- Authors
- Voleti, V., Patel, K.B., Li, W., Perez Campos, C., Bharadwaj, S., Yu, H., Ford, C., Casper, M.J., Yan, R.W., Liang, W., Wen, C., Kimura, K.D., Targoff, K.L., Hillman, E.M.C.
- ID
- ZDB-PUB-190929-14
- Date
- 2019
- Source
- Nature Methods 16: 1054-1062 (Journal)
- Registered Authors
- Targoff, Kimara
- Keywords
- none
- MeSH Terms
-
- Animals
- Caenorhabditis elegans/metabolism
- Calcium/metabolism
- Heart/embryology
- Heart/physiology
- Microscopy/methods*
- Photons
- Zebrafish/embryology
- Zebrafish/metabolism
- Zebrafish/physiology
- PubMed
- 31562489 Full text @ Nat. Methods
Citation
Voleti, V., Patel, K.B., Li, W., Perez Campos, C., Bharadwaj, S., Yu, H., Ford, C., Casper, M.J., Yan, R.W., Liang, W., Wen, C., Kimura, K.D., Targoff, K.L., Hillman, E.M.C. (2019) Real-time volumetric microscopy of in vivo dynamics and large-scale samples with SCAPE 2.0. Nature Methods. 16:1054-1062.
Abstract
The limited per-pixel bandwidth of most microscopy methods requires compromises between field of view, sampling density and imaging speed. This limitation constrains studies involving complex motion or fast cellular signaling, and presents a major bottleneck for high-throughput structural imaging. Here, we combine high-speed intensified camera technology with a versatile, reconfigurable and dramatically improved Swept, Confocally Aligned Planar Excitation (SCAPE) microscope design that can achieve high-resolution volumetric imaging at over 300 volumes per second and over 1.2 GHz pixel rates. We demonstrate near-isotropic sampling in freely moving Caenorhabditis elegans, and analyze real-time blood flow and calcium dynamics in the beating zebrafish heart. The same system also permits high-throughput structural imaging of mounted, intact, cleared and expanded samples. SCAPE 2.0's significantly lower photodamage compared to point-scanning techniques is also confirmed. Our results demonstrate that SCAPE 2.0 is a powerful, yet accessible imaging platform for myriad emerging high-speed dynamic and high-throughput volumetric microscopy applications.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping