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A hybrid open-top light-sheet microscope for versatile multi-scale imaging of cleared tissues

Abstract

Light-sheet microscopy has emerged as the preferred means for high-throughput volumetric imaging of cleared tissues. However, there is a need for a flexible system that can address imaging applications with varied requirements in terms of resolution, sample size, tissue-clearing protocol, and transparent sample-holder material. Here, we present a ‘hybrid’ system that combines a unique non-orthogonal dual-objective and conventional (orthogonal) open-top light-sheet (OTLS) architecture for versatile multi-scale volumetric imaging. We demonstrate efficient screening and targeted sub-micrometer imaging of sparse axons within an intact, cleared mouse brain. The same system enables high-throughput automated imaging of multiple specimens, as spotlighted by a quantitative multi-scale analysis of brain metastases. Compared with existing academic and commercial light-sheet microscopy systems, our hybrid OTLS system provides a unique combination of versatility and performance necessary to satisfy the diverse requirements of a growing number of cleared-tissue imaging applications.

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Fig. 1: Hybrid OTLS microscopy.
Fig. 2: Fast meso-scale screening and targeted sub-micrometer imaging in cleared tissues.
Fig. 3: Multi-scale OTLS microscopy for quantitative analysis of brain metastases.

Data availability

The customized ZEMAX files, CAD files, list of components, and a summary of all datasets and the associated imaging parameters are available as Supplementary Data. Due to the large size of the imaging datasets collected within this manuscript, the datasets are not available in a public repository and are available from the authors upon request.

Code availability

The simulation codes used to model the lateral and axial resolution of the various microscope architectures is available on GitHub. The acquisition software code for the microscope is available from the authors upon request.

References

  1. Chung, K. et al. Structural and molecular interrogation of intact biological systems. Nature 497, 332–337 (2013).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  2. Tanaka, N. et al. Whole-tissue biopsy phenotyping of three-dimensional tumours reveals patterns of cancer heterogeneity. Nat. Biomed. Eng. 1, 796 (2017).

    CAS  PubMed  Article  Google Scholar 

  3. Richardson, D. S. & Lichtman, J. W. Clarifying tissue clearing. Cell 162, 246–257 (2015).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  4. Pan, C. et al. Shrinkage-mediated imaging of entire organs and organisms using uDISCO. Nat. Methods 13, 859–867 (2016).

    CAS  PubMed  Article  Google Scholar 

  5. Renier, N. et al. iDISCO: a simple, rapid method to immunolabel large tissue samples for volume imaging. Cell 159, 896–910 (2014).

    CAS  PubMed  Article  Google Scholar 

  6. Tainaka, K. et al. Chemical landscape for tissue clearing based on hydrophilic reagents. Cell Rep. 24, 2196–2210.e9 (2018).

    CAS  PubMed  Article  Google Scholar 

  7. Susaki, E. A. et al. Advanced CUBIC protocols for whole-brain and whole-body clearing and imaging. Nat. Protoc. 10, 1709–1727 (2015).

    CAS  PubMed  Article  Google Scholar 

  8. Chung, K. & Deisseroth, K. CLARITY for mapping the nervous system. Nat. Methods 10, 508–513 (2013).

    CAS  PubMed  Article  Google Scholar 

  9. Susaki, E. A. et al. Versatile whole-organ/body staining and imaging based on electrolyte-gel properties of biological tissues. Nat. Commun. 11, 1982 (2020).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  10. Jing, D. et al. Tissue clearing of both hard and soft tissue organs with the PEGASOS method. Cell Res. 28, 803–818 (2018).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  11. Power, R. M. & Huisken, J. A guide to light-sheet fluorescence microscopy for multiscale imaging. Nat. Methods 14, 360 (2017).

    CAS  PubMed  Article  Google Scholar 

  12. Huisken, J. & Stainier, D. Y. R. Selective plane illumination microscopy techniques in developmental biology Development 136, 1963–1975 (2009).

  13. Dodt, H.-U. et al. Ultramicroscopy: three-dimensional visualization of neuronal networks in the whole mouse brain. Nat. Methods 4, 331–336 (2007).

    CAS  PubMed  Article  Google Scholar 

  14. Migliori, B. et al. Light sheet theta microscopy for rapid high-resolution imaging of large biological samples. BMC Biol. 16, 57 (2018).

    PubMed  PubMed Central  Article  CAS  Google Scholar 

  15. Tomer, R. et al. Advanced CLARITY for rapid and high-resolution imaging of intact tissues. Nat. Protoc. 9, 1682–1697 (2014).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  16. Voigt, F. F. et al. The mesoSPIM initiative: open-source light-sheet microscopes for imaging cleared tissue. Nat. Methods 16, 1105–1108 (2019).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  17. Chakraborty, T. et al. Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution. Nat. Methods 16, 1109–1113 (2019).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  18. Chen, Y. et al. A versatile tiling light sheet microscope for imaging of cleared tissues. Cell Rep. 33, 108349 (2020).

  19. Kumar, A. et al. Dual-view plane illumination microscopy for rapid and spatially isotropic imaging. Nat. Protoc. 9, 2555–2573 (2014).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  20. Strnad, P. et al. Inverted light-sheet microscope for imaging mouse pre-implantation development. Nat. Methods 13, 139–142 (2015).

    PubMed  Article  CAS  Google Scholar 

  21. McGorty, R. et al. Open-top selective plane illumination microscope for conventionally mounted specimens. Opt. Express 23, 16142–16153 (2015).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  22. McGorty, R., Xie, D. & Huang, B. High-NA open-top selective-plane illumination microscopy for biological imaging. Opt. Express 25, 17798–17810 (2017).

    PubMed  PubMed Central  Article  Google Scholar 

  23. Glaser, A. K. et al. Light-sheet microscopy for slide-free non-destructive pathology of large clinical specimens. Nat. Biomed. Eng. 1, 0084 (2017).

    PubMed  PubMed Central  Article  Google Scholar 

  24. Glaser, A. K. et al. Multi-immersion open-top light-sheet microscope for high-throughput imaging of cleared tissues. Nat. Commun. 10, 1–8 (2019).

    Article  Google Scholar 

  25. Barner, L. A. et al. Solid immersion meniscus lens (SIMlens) for open-top light-sheet microscopy. Opt. Lett. 44, 4451–4454 (2019).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  26. Barner, L. A. et al. Multi-resolution open-top light-sheet microscopy to enable efficient 3D pathology workflows. Biomed. Opt. Express 11, 6605–6619 (2020).

    PubMed  PubMed Central  Article  Google Scholar 

  27. Botcherby, E. J. et al. An optical technique for remote focusing in microscopy. Opt. Commun. 281, 880–887 (2008).

    CAS  Article  Google Scholar 

  28. Dunsby, C. Optically sectioned imaging by oblique plane microscopy. Opt. Express 16, 20306–20316 (2008).

    CAS  PubMed  Article  Google Scholar 

  29. Voleti, V. et al. Real-time volumetric microscopy of in vivo dynamics and large-scale samples with SCAPE 2.0. Nat. Methods 16, 1054–1062 (2019).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  30. Bouchard, M. B. et al. Swept confocally-aligned planar excitation (SCAPE) microscopy for high-speed volumetric imaging of behaving organisms. Nat. Photonics 9, 113–119 (2015).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  31. Yang, B. et al. Epi-illumination SPIM for volumetric imaging with high spatial-temporal resolution. Nat. Methods 16, 501–504 (2019).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  32. Millett-Sikking, A. et al. High NA single-objective light-sheet. https://andrewgyork.github.io/high_na_single_objective_lightsheet/index.html (2019).

  33. Kumar, M. et al. Integrated one- and two-photon scanned oblique plane illumination (SOPi) microscopy for rapid volumetric imaging. Opt. Express 26, 13027–13041 (2018).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  34. Hoffmann, M. & Judkewitz, B. Diffractive oblique plane microscopy. Optica 6, 5 (2019).

    Article  Google Scholar 

  35. Sapoznik, E. et al. A versatile oblique plane microscope for large-scale and high-resolution imaging of subcellular dynamics. eLife 9, e57681 (2020).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  36. Yang, B. et al. DaXi-high-resolution, large imaging volume and multi-view single-objective light-sheet microscopy. Nat. Methods 19, 461–469 (2022).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  37. Li, T. et al. Axial plane optical microscopy. Sci. Rep. 4, 7253 (2014).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  38. Kim, J. et al. Oblique-plane single-molecule localization microscopy for tissues and small intact animals. Nat. Methods 16, 853–857 (2019).

    CAS  PubMed  Article  Google Scholar 

  39. Bishop, K. W., Glaser, A. K. & Liu, J. T. C. Performance tradeoffs for single- and dual-objective open-top light-sheet microscope designs: a simulation-based analysis. Biomed. Opt. Express 11, 4627–4650 (2020).

    PubMed  PubMed Central  Article  Google Scholar 

  40. Economo, M. N. et al. A platform for brain-wide imaging and reconstruction of individual neurons. eLife 5, e10566 (2016).

    PubMed  PubMed Central  Article  Google Scholar 

  41. Winnubst, J. et al. Reconstruction of 1,000 projection neurons reveals new cell types and organization of long-range connectivity in the mouse brain. Cell 179, 268–281.e13 (2019).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  42. Kubota, S. I. et al. Whole-body profiling of cancer metastasis with single-cell resolution. Cell Rep. 20, 236–250 (2017).

    CAS  PubMed  Article  Google Scholar 

  43. Planchon, T. A. et al. Rapid three-dimensional isotropic imaging of living cells using Bessel beam plane illumination. Nat. Methods 8, 417–423 (2011).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  44. Dean, K. et al. Deconvolution-free subcellular imaging with axially swept light sheet microscopy. Biophys. J. 108, 2807–2815 (2015).

  45. Sparks, H. et al. Dual-view oblique plane microscopy (dOPM). Biomed. Opt. Express 11, 7204–7220 (2020).

    Article  Google Scholar 

  46. Keller, P. J. et al. Fast, high-contrast imaging of animal development with scanned light sheet-based structured-illumination microscopy. Nat. Methods 7, 637–642 (2010).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  47. Yueqian, Z. & Herbert, G. Systematic design of microscope objectives. Part I: system review and analysis. Adv. Opt. Technol. 8, 313–347 (2019).

    Article  Google Scholar 

  48. Hörl, D. et al. BigStitcher: reconstructing high-resolution image datasets of cleared and expanded samples. Nat. Methods 16, 870–874 (2019).

    PubMed  Article  CAS  Google Scholar 

  49. Schindelin, J. et al. Fiji: an open-source platform for biological-image analysis. Nat. Methods 9, 676–682 (2012).

    CAS  PubMed  Article  Google Scholar 

  50. Balazs, B., et al. A real-time compression library for microscopy images. Preprint at bioRxiv https://doi.org/10.1101/164624 (2017).

  51. Chan, K. Y. et al. Engineered AAVs for efficient noninvasive gene delivery to the central and peripheral nervous systems. Nat. Neurosci. 20, 1172–1179 (2017).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  52. Chi, J. et al. Three-dimensional adipose tissue imaging reveals regional variation in beige fat biogenesis and PRDM16-dependent sympathetic neurite density. Cell Metab. 27, 226–236.e3 (2018).

    CAS  PubMed  Article  Google Scholar 

  53. Ehata, S. et al. Transforming growth factor-β promotes survival of mammary carcinoma cells through induction of antiapoptotic transcription factor DEC1. Cancer Res. 67, 9694 (2007).

    CAS  PubMed  Article  Google Scholar 

  54. Nishida, J. et al. Epigenetic remodelling shapes inflammatory renal cancer and neutrophil-dependent metastasis. Nat. Cell Biol. 22, 465–475 (2020).

    CAS  PubMed  Article  Google Scholar 

  55. Miyoshi, H., Takahashi, M., Gage, F. H. & Verma, I. M. Stable and efficient gene transfer into the retina using an HIV-based lentiviral vector. Proc. Natl Acad. Sci. USA 94, 10319–10323 (1997).

    CAS  PubMed  PubMed Central  Article  Google Scholar 

  56. Matsumoto, K. et al. Advanced CUBIC tissue clearing for whole-organ cell profiling. Nat. Protoc. 14, 3506–3537 (2019).

    CAS  PubMed  Article  Google Scholar 

Download references

Acknowledgements

We would like to thank A. York and A. Millet-Sikking for discussions regarding oblique planar microscopy, remote focus imaging, and alignment procedures. We would also like to thank J. Daniels for discussions and his development of the multi-immersion objective, C. Shimizu (RIKEN BDR) for the support of preparing CUBIC-cleared and stained specimens, and K. Miyazono (The University of Tokyo) for the discussion and support of brain metastasis experiments. This work was funded in part by the National Institutes of Health (NIH) K99 CA240681 (A.K.G.), R01CA244170 (J.T.C.L.), R01EB031002 (J.T.C.L.), R01GM079712 (T.I.), R01DK107436 (L.X.), R01DK092202 (L.X.), R01MH117820 and R01NS104949 (R.C.R.); Department of Defense (DoD) Prostate Cancer Research Program (PCRP) W81XWH-18-10358 (J.T.C.L. and L.D.T.), W81XWH-19-1-0589 (N.P.R.), W81XWH-20-1-0039 (X.W.), and Prostate Cancer Young Investigator Award (N.P.R.); National Science Foundation (NSF) Graduate Research Fellowship DGE-1762114 (L.A.B. and K.W.B.); NSF 1934292 HDR: I-DIRSE-FW (J.T.C.L. and R.B.S.); Washington Research Foundation Postdoctoral Fellowship (C.R.S.); Science and Technology Platform Program for Advanced Biological Medicine by the Japan Agency for Medical Research and Development (AMED) JP21am0401011 (H.R.U.), ERATO by Japan Science and Technology Agency (JST) JPMJER2001 (H.R.U.), HFSP Research Grant Program RGP0019/2018 (H.R.U.), AMED-PRIME JP21gm6210027 (E.A.S.), Grants-in-Aid for Scientific Research on Innovative Areas (JSPS KAKENHI grant) 17H06328 (E.A.S.), Grants-in-Aid for Scientific Research on Innovative Areas (JSPS KAKENHI grant) 20K1612 (S.I.K.). Work in the Murawala laboratory is supported by grants from NIH-COBRE (5P20GM104318-08) and DFG (429469366). Work in the Dodt laboratory is supported by grants FWF P31263-B26 and WWTF CS19-019.

Author information

Authors and Affiliations

Authors

Contributions

A.K.G. and J.T.C.L. conceived of and designed the microscope system. P.R.N. provided feedback on the system design and its potential applications. A.K. G., K.W.B., R.B.S., and G.G. performed simulations of the microscope. A.K.G. fabricated the microscope system with help from L.A.B.E.A.S. and H.R.U. provided and prepared the immunostained CUBIC-cleared mouse brains. E.A.S., S.I.K., and H.R.U. prepared and provided the metastatic mouse brains. J.C. and K.S. prepared and provided the mouse brain. P.B., E.T., and R.C.R. provided the mouse brain with preparation by A.K.G. The human brain slice was prepared by A.K.G.H.L. and L.A.G.L. provided quantitative analysis of the high-resolution images. Y.Y. and H.Z. provided and prepared the PEGASOS-cleared mouse brain. E.K.N., B.J.B., and J.S. provided and prepared the SHIELD-cleared mouse embryos. H.H., N.P.R., and L.D.T. provided and prepared the ECi-cleared human prostate tissue. J.J.W., R.S., E.S., C.R.S., and M.Y.G. provided and prepared the Ce3D-cleared mouse lymph node. X.W. and L.X. provided and prepared the iDISCO-cleared mouse prostate. A.K.H. and T.I. provided and prepared the ClearSee-processed Arabidopsis plant and M.P., P.M., and H.U.D. provided and prepared the DEEP-Clear-processed Axolotl. A.K.G. and J.T.C.L. led the writing of the manuscript. All authors contributed to the manuscript.

Corresponding authors

Correspondence to Adam K. Glaser or Jonathan T. C. Liu.

Ethics declarations

Competing interests

A.K.G., N.P.R., L.D.T., and J.T.C.L. are co-founders and shareholders of Alpenglow Biosciences. L.A.G.L. and H.L. are employees of Leica Microsystems, maker of the Aivia software.

Peer review

Peer review information

Nature Methods thanks Peter Santi, Per Uhlen, and Fabian Voigt for their contribution to the peer review of this work. Rita Strack was the primary editor on this article and managed its editorial process and peer review in collaboration with the rest of the editorial team.

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Extended data

Extended Data Fig. 1 Regions of interests from metastatic brain specimens.

All n = 34 metastatic colonies collected from the metastatic brain specimens (3 brains from OS-RC-2 cancer cell line, 3 brains from MDA-MB-231 cancer cell line). All scale bars represent 10 μm.

Extended Data Fig. 2 Multi-scale 3D pathology of human prostate tissue.

(a) Fast meso-scale screening results for a multi-cm-sized piece of human prostate tissue stained with TO-PRO-3 Iodide (nuclear) and Eosin. Representative high-resolution regions of interests for two different foci of cancer are shown in (b) and (c). The insets demonstrate the ability to clearly resolve sub-nuclear features in cancer nuclei. Scale bar lengths are as follows: (a) 1 cm, (b) and (c) 10 μm. The imaging data in (a-c) was acquired from in a single experiment, which was not repeated.

Extended Data Fig. 3 Assessment of 3D cell proliferation assays with iDISCO.

(a) Screening of an intact mouse prostate cleared using iDISCO, labeled with TOPRO3 Iodide (nuclear) and an EduClick cell-proliferation marker. A higher-resolution region of interest focused on a prostate gland is shown in (b). An additional zoomed-in view showing the ability to resolve individual proliferating nuclei (c). Scale bars lengths are as follows: (a) 1 mm, (b) 200 μm, and (c) 100 μm. The imaging data in (a-c) was acquired from in a single experiment, which was not repeated.

Extended Data Fig. 4 Imaging of non-rodent and non-human tissues.

(a) Hybrid open-top light-sheet imaging of a ClearSee-cleared Arabidopsis specimen. (b) Higher-resolution imaging of the Arabidopsis root. (c) Meso-scale imaging of a large multi-cm Axolotl cleared with DEEP-Clear. Scale bars lengths are as follows: (a) 1 mm, (b) 100 μm, and (c) 1 cm. The imaging data in (a-c) was acquired from in a single experiment, which was not repeated.

Extended Data Fig. 5 Imaging of immunostained and endogenously fluorescent CUBIC-cleared mouse brains.

Hybrid OTLS imaging of whole mouse brains. (a-c) with endogenously preserved GCaMP6s fluorescence, and immunostained with (d) anti-ChAT antibody + SYTOX-G or (e) anti-Parvalbumin (PV) antibody + SYTOX-G by CUBIC-HistoVision. Scale bar lengths are as follows: (a-b) 1 mm, (c) 10 μm, (d-e) 2 mm. The imaging data in (a-e) was acquired from in a single experiment, which was not repeated.

Extended Data Fig. 6 Large-scale imaging of thick human tissues.

(a) Slab of human brain tissue after CUBIC clearing. A mouse brain is shown for size comparison. (b) ODO imaging results of the entire 3-mm thick brain slab. Autofluorescence is shown in black and white, and the amyloid small molecule stain (pFTAA) is shown in green. (c) Zoom-in views of an amyloid-rich region reveal perivascular accumulation. Scale bar lengths are as follows: (b) 1 cm. The imaging data in (a-c) was acquired from in a single experiment, which was not repeated.

Extended Data Fig. 7 NODO imaging quality versus imaging depth in a CUBIC-cleared mouse brain.

(a) Multiple two-channel regions of interest (ROIs) were acquired from a CUBIC-cleared mouse brain at various imaging depths (z = 1–8 mm). (b-e) xy views for ROIs at various depths are shown. The corresponding xz views of each ROI are shown in (f-i). The high-magnification insets show fine sub-nuclear details and demonstrate that there is minimal degradation in image quality as a function of depth. All scalebars denote 10 µm. The imaging data in (b-i) was acquired from in a single experiment, which was not repeated.

Extended Data Fig. 8 NODO imaging quality versus imaging depth in a PEGASOS-cleared mouse brain.

(a) A single ROI was acquired from a PEGASOS-cleared mouse brain. (b) xz view of the ~5 mm deep ROI, demonstrating near-consistent image quality versus depth. (c-g) xy views for ROIs at various depths are shown. The high-magnification insets show fine sub-nuclear details and demonstrate that there is minimal degradation in image quality as a function of depth. Scale bar lengths are as follows: (b) 100 µm, (c-g) 10 µm. The imaging data in (b-g) was acquired from in a single experiment, which was not repeated.

Extended Data Fig. 9 NODO imaging quality versus imaging depth in an ECi-cleared mouse brain.

(a) Multiple regions of interest (ROIs) were acquired from an ECi-cleared mouse brain at various imaging depths (z = 0–5 mm). (b-f) xy views for ROIs at various depths are shown. The corresponding xz views of each ROI are shown in (g-k). The high-magnification insets show fine sub-nuclear details and demonstrate that there is minimal degradation in image quality as a function of depth. All scalebars denote 10 µm. The imaging data in (b-k) was acquired from in a single experiment, which was not repeated.

Supplementary information

Supplementary Information

Supplementary Notes 1–6, Supplementary Methods, Supplementary Tables 1–3, Supplementary Figures 1–25, References

Reporting Summary

Supplementary Video 1

Live video showing assembly and operation of the hybrid open-top light-sheet microscope.

Supplementary Video 2

ODO and NODO imaging results from a CUBIC-cleared mouse brain stained with SYTOX Green and αSMA.

Supplementary Video 3

Sparse axonal imaging results from an ECI-cleared Slc17a7-Cre mouse brain.

Supplementary Video 4

Multi-scale imaging of MDA-MB-231 and OS-RC-2 metastatic lesions in CUBIC-cleared mouse brains.

Supplementary Video 5

ODO and NODO imaging results from a PEGASOS-cleared Thy1-EGFP mouse brain.

Supplementary Video 6

Multi-channel 3D imaging of a Ce3D-cleared mouse lymph node.

Supplementary Video 7

Assessment of 3D cell proliferation in an iDISCO-cleared mouse prostate.

Supplementary Video 8

ODO imaging results from CUBIC-HV cleared and stained mouse brains.

Supplementary Video 9

Imaging results from a CUBIC-cleared Slc17a7-IRES2-Cre;Ai14 mouse brain.

Supplementary Video 10

Imaging results from a CUBIC-cleared Chat-IRES-Cre;Ai162 mouse brain.

Supplementary Video 11

Large-scale imaging of a thick CUBIC-cleared human brain slab.

Supplementary Data

ZEMAX files for the hybrid open-top light-sheet microscope

Supplementary Data

CAD files for the hybrid open-top light-sheet microscope

Supplementary Data

Parts list for the hybrid open-top light-sheet microscope

Supplementary Data

Summary of experimental imaging parameters for datasets

Supplementary Data

Specifications of existing immersion objective lenses

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Glaser, A.K., Bishop, K.W., Barner, L.A. et al. A hybrid open-top light-sheet microscope for versatile multi-scale imaging of cleared tissues. Nat Methods 19, 613–619 (2022). https://doi.org/10.1038/s41592-022-01468-5

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