Whole-brain optical access in a small adult vertebrate with two- and three-photon microscopy

Akbari, Najva; Tatarsky, Rose L.; Kolkman, Kristine E.; Fetcho, Joseph R.; Bass, Andrew H.; Xu, Chris

doi:10.1016/j.isci.2022.105191

Cited by 16 publications

(13 citation statements)

References 41 publications

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“…(2023). Non‐invasive multiphoton microscopy further enables long‐term studies over days to weeks in the same animal, as we showed earlier for D. dracula (Akbari et al., 2022). The ability of THG microscopy to resolve myelinated structures, blood vessels, and neuronal cell bodies throughout the brain without the need for any biomarkers could allow unbiased identification of changes in brain regions (e.g., telencephalon, optic tectum, and cerebellum) that exhibit increased myelination during critical developmental timepoints (Makinodan et al., 2012) reflecting enhanced connectivity of neuronal networks that underlie adult behaviors (Halbach & Nixdorf‐Bergweiler, 2004; Hamaide et al., 2017).…”

Section: Discussionmentioning

confidence: 54%

“…Imaging methods were largely adopted from previous studies (Akbari et al., 2022; Chow et al., 2020). The amplitude of the THG signal is strongly dependent on the numerical aperture, laser repetition rate, and pulse length (Cheng et al., 2014).…”

Section: Methodsmentioning

confidence: 99%

“…We recently demonstrated THG imaging of vasculature at all depths of the adult brain in species within the genus Danionella (Akbari et al, 2022;Chow et al, 2020), which are a miniature, translucent teleost fish closely related to zebrafish (Figure 1) (Britz et al, 2009;Conway et al, 2021;Hoffmann et al, 2023;Lam, 2022;Lindemann et al, 2022;Penalva et al, 2018;Schulze et al, 2018). This included visualizing brain vasculature in the same brain region and individual at different time points over nearly 2 months (Akbari et al, 2022). Here, we demonstrate label-free mapping with THG microscopy of neuronal populations, individual neurons, and fiber bundles in multiple regions of an entire adult Danionella brain.…”

mentioning

confidence: 99%

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Label‐free, whole‐brain in vivo mapping in an adult vertebrate with third harmonic generation microscopy

Akbari,

Tatarsky,

Kolkman

et al. 2024

J of Comparative Neurology

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Comprehensive understanding of interconnected networks within the brain requires access to high resolution information within large field of views and over time. Currently, methods that enable mapping structural changes of the entire brain in vivo are extremely limited. Third harmonic generation (THG) can resolve myelinated structures, blood vessels, and cell bodies throughout the brain without the need for any exogenous labeling. Together with deep penetration of long wavelengths, this enables in vivo brain‐mapping of large fractions of the brain in small animals and over time. Here, we demonstrate that THG microscopy allows non‐invasive label‐free mapping of the entire brain of an adult vertebrate, Danionella dracula, which is a miniature species of cyprinid fish. We show this capability in multiple brain regions and in particular the identification of major commissural fiber bundles in the midbrain and the hindbrain. These features provide readily discernable landmarks for navigation and identification of regional‐specific neuronal groups and even single neurons during in vivo experiments. We further show how this label‐free technique can easily be coupled with fluorescence microscopy and used as a comparative tool for studies of other species with similar body features to Danionella, such as zebrafish (Danio rerio) and tetras (Trochilocharax ornatus). This new evidence, building on previous studies, demonstrates how small size and relative transparency, combined with the unique capabilities of THG microscopy, can enable label‐free access to the entire adult vertebrate brain.

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Section: Discussionmentioning

confidence: 54%

Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

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Label‐free, whole‐brain in vivo mapping in an adult vertebrate with third harmonic generation microscopy

Akbari,

Tatarsky,

Kolkman

et al. 2024

J of Comparative Neurology

Self Cite

View full text Add to dashboard Cite

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“…Combined with cutting-edge optical technologies, such as adaptive excitation sources (AES) ( Li et al, 2020 ), adaptive optics (AO) ( Rodríguez et al, 2021 ; Streich et al, 2021 ; Qin et al, 2022 ; Sinefeld et al, 2022 ), and Bessel beam ( Chen et al, 2018 ; Rodríguez et al, 2018 ), 3PM has made significant advances in imaging performance. In application scenarios, increasing research groups have demonstrated the capabilities of 3PM imaging for probing neural structural and functional dynamics beyond the depth of 2PM, including the applications of 3PM in imaging rodents ( Horton et al, 2013 ; Ouzounov et al, 2017 ; Wang T. et al, 2018 ; Liu et al, 2019a ; Weisenburger et al, 2019 ; Klioutchnikov et al, 2020 , 2022 ; Wang et al, 2021 ; Choe et al, 2022 ), fishes ( Chow et al, 2020 ; Rodríguez et al, 2021 ; Akbari et al, 2022a ), flies ( Tao et al, 2017 ; Hsu et al, 2019 ; Aragon et al, 2022 ), and other kinds of tissues ( He et al, 2019 ; Yildirim et al, 2020 ; Wang et al, 2021 ).…”

Section: Introductionmentioning

confidence: 99%

Three-photon excited fluorescence imaging in neuroscience: From principles to applications

Xiao

Deng²,

Zhao³

et al. 2023

Front. Neurosci.

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The development of three-photon microscopy (3PM) has greatly expanded the capability of imaging deep within biological tissues, enabling neuroscientists to visualize the structure and activity of neuronal populations with greater depth than two-photon imaging. In this review, we outline the history and physical principles of 3PM technology. We cover the current techniques for improving the performance of 3PM. Furthermore, we summarize the imaging applications of 3PM for various brain regions and species. Finally, we discuss the future of 3PM applications for neuroscience.

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“…To improve the imaging depth, an effective strategy is using longer excitation wavelength to reduce the attenuation of excitation light through tissues. [1,2,16,25,26] Typically, four "tissue optical windows" are recognized and proved to be potential for deep-brain vascular imaging: NIR-I (800 nm window, 650-950 nm), [14,[25][26][27] NIR-II (1300 nm window, 1100-1350 nm), [6,[27][28][29][30] NIR-III (1700 nm window, 1600-1840 nm), [1,2,[31][32][33][34] and NIR-IV (2200 nm window, 2100-2300 nm). [35][36][37][38] Using 775 nm excitation indicated that the maximum 2PF imaging depth of brain blood vessels is ≈650 μm below the mouse brain surface, which is mainly limited by the SBR.…”

Section: Introductionmentioning

confidence: 99%

De Novo Design of Aggregation‐Induced Emission Luminogen for Three‐Photon Fluorescence Imaging of Subcortical Structures Excited at Both NIR‐III and NIR‐IV Windows

Tong

Zhong

et al. 2023

Adv Funct Materials

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Three‐photon fluorescence (3PF) imaging excited at 1700 nm window is an enabling technology for visualizing deep brain structures and dynamics. Recently, the 2200 nm window has emerged as the longest excitation window suitable for deep‐brain 3PF imaging. Bright fluorescent probes lay the material basis for deep‐brain 3PF imaging. Among various fluorescent probes, aggregation‐induced emission luminogens (AIEgens) have great potential in 3PF imaging excited at the 1700 nm window in vivo. However, to the best of knowledge, there is no AIEgens applicable to 3PF imaging excited at both the 1700 and 2200 nm windows. To readily fill this gap, here this study designs and synthesizes a novel AIEgen, namely TPE‐DPTT‐ICP, which generates bright 3PF signals excited at both 1700 and 2200 nm. The accordingly fabricated TPE‐DPTT‐ICP nanoparticles (NPs) possess excellent water dispersibility, colloidal stability, biocompatibility, photostability and large 3P action cross section, key to in vivo imaging. In mouse brain in vivo, TPE‐DPTT‐ICP NPs enable deep‐brain 3PF imaging of subcortical structures excited at both the two windows, reaching depths of 1640 and 880 µm below the brain surface, respectively. TPE‐DPTT‐ICP NPs are thus a versatile material simultaneously catering to the need at two infrared optical windows with deep tissue penetration.

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Whole-brain optical access in a small adult vertebrate with two- and three-photon microscopy

Cited by 16 publications

References 41 publications

Label‐free, whole‐brain in vivo mapping in an adult vertebrate with third harmonic generation microscopy

Label‐free, whole‐brain in vivo mapping in an adult vertebrate with third harmonic generation microscopy

Three-photon excited fluorescence imaging in neuroscience: From principles to applications

De Novo Design of Aggregation‐Induced Emission Luminogen for Three‐Photon Fluorescence Imaging of Subcortical Structures Excited at Both NIR‐III and NIR‐IV Windows

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