Light sheet theta microscopy for rapid high-resolution imaging of large biological samples
BMC biology, 2018•Springer
Background Advances in tissue clearing and molecular labeling methods are enabling
unprecedented optical access to large intact biological systems. These developments fuel
the need for high-speed microscopy approaches to image large samples quantitatively and
at high resolution. While light sheet microscopy (LSM), with its high planar imaging speed
and low photo-bleaching, can be effective, scaling up to larger imaging volumes has been
hindered by the use of orthogonal light sheet illumination. Results To address this …
unprecedented optical access to large intact biological systems. These developments fuel
the need for high-speed microscopy approaches to image large samples quantitatively and
at high resolution. While light sheet microscopy (LSM), with its high planar imaging speed
and low photo-bleaching, can be effective, scaling up to larger imaging volumes has been
hindered by the use of orthogonal light sheet illumination. Results To address this …
Background
Advances in tissue clearing and molecular labeling methods are enabling unprecedented optical access to large intact biological systems. These developments fuel the need for high-speed microscopy approaches to image large samples quantitatively and at high resolution. While light sheet microscopy (LSM), with its high planar imaging speed and low photo-bleaching, can be effective, scaling up to larger imaging volumes has been hindered by the use of orthogonal light sheet illumination.
Results
To address this fundamental limitation, we have developed light sheet theta microscopy (LSTM), which uniformly illuminates samples from the same side as the detection objective, thereby eliminating limits on lateral dimensions without sacrificing the imaging resolution, depth, and speed. We present a detailed characterization of LSTM, and demonstrate its complementary advantages over LSM for rapid high-resolution quantitative imaging of large intact samples with high uniform quality.
Conclusions
The reported LSTM approach is a significant step for the rapid high-resolution quantitative mapping of the structure and function of very large biological systems, such as a clarified thick coronal slab of human brain and uniformly expanded tissues, and also for rapid volumetric calcium imaging of highly motile animals, such as Hydra, undergoing non-isomorphic body shape changes.
Springer
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