Temporally multiplexed dual-plane imaging of neural activity with four-dimensional precision. (October 2021)

• Record Type: Journal Article
• Authors: Onda, Masanari; Takeuchi, Ryosuke F.; Isobe, Keisuke; Suzuki, Toshiaki; Masaki, Yuji; Morimoto, Nao; Osakada, Fumitaka
• Is Part Of: Neuroscience research. Volume 171(2021)
• Page Start: 9
• Page End: 18
• Publication Date: 2021-10
• Subjects: Multiplane imaging -- Neural activity -- Neural circuit -- Remote focusing -- Spatial light modulator -- Two-photon microscope -- Viral vector; Neurosciences -- Research -- Periodicals; Neurosciences -- Research -- Japan -- Periodicals; Neurology -- Periodicals; Neurosciences -- Periodicals; Neurosciences -- Recherche -- Périodiques; Neurosciences -- Recherche -- Japon -- Périodiques; Neurosciences -- Research; Japan; Periodicals; 612.8
• DOI: 10.1016/j.neures.2021.02.001
• Languages: English
• ISSNs: 0168-0102

Abstract

Highlights: A holographic two-photon microscope was developed for dual-plane imaging. A spatial light modulator generated an additional focal plane. A temporal multiplexing strategy allowed fast imaging of two different focal planes. 4-D imaging was achieved at cellular resolution and physiological timescales. 4-D imaging and analysis will facilitate studies linking structure and function. Abstract: Spatiotemporal patterns of neural activity generate brain functions, such as perception, memory, and behavior. Four-dimensional (4-D: x, y, z, t ) analyses of such neural activity will facilitate understanding of brain functions. However, conventional two-photon microscope systems observe single-plane brain tissue alone at a time with cellular resolution. It faces a trade-off between the spatial resolution in the x -, y -, and z -axes and the temporal resolution by a limited point-by-point scan speed. To overcome this trade-off in 4-D imaging, we developed a holographic two-photon microscope for dual-plane imaging. A spatial light modulator (SLM) provided an additional focal plane at a different depth. Temporal multiplexing of split lasers with an optical chopper allowed fast imaging of two different focal planes. We simultaneously recorded the activities of neurons on layers 2/3 and 5 of the cerebral cortex in awake mice in vivo . The present study demonstrated the proof-of-concept of dual-plane two-photon imaging of neural circuits by using the temporally multiplexed SLM-based microscope. The temporally multiplexed holographic microscope, combined with in vivo labeling with genetically encoded probes, enabled 4-D imaging and analysis of neural activities at cellular resolution and physiological timescales. Large-scale 4-D imaging and analysis will facilitate studies of not only the nervous system but also of various biological systems.