Axially Swept Light-Sheet Microscopy (ASLM)

Only two light-sheet microscopy techniques successfully decouple the trade-off between axial resolution and field of view: multiview deconvolution and ASLM. In the former, the sample is imaged from multiple directions (via sample rotation or by reversing the illumination and detection paths) and the complementary views computationally fused to create a high-resolution, large field of view image. Thus, the amount of data produced, which can be immense to begin with, is effectively tripled. In contrast, ASLM, a technology invented by Dean in 2015, uses aberration-free remote focusing to sweep a tightly focused Gaussian beam through the sample in its propagation direction synchronously with the rolling shutter of a CMOS camera. Here, the axial position of the illumination beam is deterministically controlled by translating the position of a piezo or voice coil mounted mirror that is optically conjugate to the specimen at the focal plane of a remote focusing objective. Importantly, ASLM does not require any computational post-processing, and delivers a large field of view (100 x 100 x 100μm) and the highest axial resolution (~400 nm) in LSFM in the absence of deconvolution or super-resolution techniques (e.g., structured illumination, stimulated emission, or localization).

Publications

  1. Zhou FY, Weems A, Gihana GM, Chen B, Chang BJ, Driscoll M, Danuser G. Surface-guided computing to analyze subcellular morphology and membrane-associated signals in 3D. bioRxiv [Preprint]. 2023 Apr 20:2023.04.12.536640. doi: 10.1101/2023.04.12.536640. PMID: 37131779; PMCID: PMC10153113.

  2. Weems AD, Welf ES, Driscoll MK, Zhou FY, Mazloom-Farsibaf H, Chang BJ, Murali VS, Gihana GM, Weiss BG, Chi J, Rajendran D, Dean KM, Fiolka R, Danuser G. Blebs promote cell survival by assembling oncogenic signalling hubs. Nature. 2023 Mar;615(7952):517-525. doi: 10.1038/s41586-023-05758-6. Epub 2023 Mar 1. PubMed PMID: 36859545.

  3. Dean KM, Chakraborty T, Daetwyler S, Lin J, Garrelts G, M'Saad O, Mekbib HT, Voigt FF, Schaettin M, Stoeckli ET, Helmchen F, Bewersdorf J, Fiolka R. Isotropic imaging across spatial scales with axially swept light-sheet microscopy. Nat Protoc. 2022 Sep;17(9):2025-2053. doi: 10.1038/s41596-022-00706-6. Epub 2022 Jul 13. Review. PubMed PMID: 35831614; PubMed Central PMCID: PMC10111370.

  4. Liu H, Hiremath C, Patterson Q, Vora S, Shang Z, Jamieson AR, Fiolka R, Dean KM, Dellinger MT, Marciano DK. Heterozygous Mutation of Vegfr3 Reduces Renal Lymphatics without Renal Dysfunction. J Am Soc Nephrol. 2021 Dec 1;32(12):3099-3113. doi: 10.1681/ASN.2021010061. Epub 2021 Dec 1. PubMed PMID: 34551997; PubMed Central PMCID: PMC8638391.

  5. Welf ES, Miles CE, Huh J, Sapoznik E, Chi J, Driscoll MK, Isogai T, Noh J, Weems AD, Pohlkamp T, Dean K, Fiolka R, Mogilner A, Danuser G. Actin-Membrane Release Initiates Cell Protrusions. Dev Cell. 2020 Dec 21;55(6):723-736.e8. doi: 10.1016/j.devcel.2020.11.024. Epub 2020 Dec 11. PubMed PMID: 33308479; PubMed Central PMCID: PMC7908823.

  6. Chakraborty T, Chen B, Daetwyler S, Chang BJ, Vanderpoorten O, Sapoznik E, Kaminski CF, Knowles TPJ, Dean KM, Fiolka R. Converting lateral scanning into axial focusing to speed up three-dimensional microscopy. Light Sci Appl. 2020;9:165. doi: 10.1038/s41377-020-00401-9. eCollection 2020. PubMed PMID: 33024553; PubMed Central PMCID: PMC7501866.

  7. Chakraborty T, Driscoll MK, Jeffery E, Murphy MM, Roudot P, Chang BJ, Vora S, Wong WM, Nielson CD, Zhang H, Zhemkov V, Hiremath C, De La Cruz ED, Yi Y, Bezprozvanny I, Zhao H, Tomer R, Heintzmann R, Meeks JP, Marciano DK, Morrison SJ, Danuser G, Dean KM, Fiolka R. Light-sheet microscopy of cleared tissues with isotropic, subcellular resolution. Nat Methods. 2019 Nov;16(11):1109-1113. doi: 10.1038/s41592-019-0615-4. Epub 2019 Oct 31. PubMed PMID: 31673159; PubMed Central PMCID: PMC6924633.

  8. Driscoll MK, Welf ES, Jamieson AR, Dean KM, Isogai T, Fiolka R, Danuser G. Robust and automated detection of subcellular morphological motifs in 3D microscopy images. Nat Methods. 2019 Oct;16(10):1037-1044. doi: 10.1038/s41592-019-0539-z. Epub 2019 Sep 9. PubMed PMID: 31501548; PubMed Central PMCID: PMC7238333.

  9. Renkawitz J, Kopf A, Stopp J, de Vries I, Driscoll MK, Merrin J, Hauschild R, Welf ES, Danuser G, Fiolka R, Sixt M. Nuclear positioning facilitates amoeboid migration along the path of least resistance. Nature. 2019 Apr;568(7753):546-550. doi: 10.1038/s41586-019-1087-5. Epub 2019 Apr 3. PMID: 30944468; PMCID: PMC7217284.

  10. Mohan AS, Dean KM, Isogai T, Kasitinon SY, Murali VS, Roudot P, Groisman A, Reed DK, Welf ES, Han SJ, Noh J, Danuser G. Enhanced Dendritic Actin Network Formation in Extended Lamellipodia Drives Proliferation in Growth-Challenged Rac1(P29S) Melanoma Cells. Dev Cell. 2019 May 6;49(3):444-460.e9. doi: 10.1016/j.devcel.2019.04.007. PubMed PMID: 31063759; PubMed Central PMCID: PMC6760970.

  11. Chu J, Oh Y, Sens A, Ataie N, Dana H, Macklin JJ, Laviv T, Welf ES, Dean KM, Zhang F, Kim BB, Tang CT, Hu M, Baird MA, Davidson MW, Kay MA, Fiolka R, Yasuda R, Kim DS, Ng HL, Lin MZ. A bright cyan-excitable orange fluorescent protein facilitates dual-emission microscopy and enhances bioluminescence imaging in vivo. Nat Biotechnol. 2016 Jul;34(7):760-7. doi: 10.1038/nbt.3550. Epub 2016 May 30. PubMed PMID: 27240196; PubMed Central PMCID: PMC4942401.

  12. Dean KM, Roudot P, Welf ES, Danuser G, Fiolka R. Deconvolution-free Subcellular Imaging with Axially Swept Light Sheet Microscopy. Biophys J. 2015 Jun 16;108(12):2807-15. doi: 10.1016/j.bpj.2015.05.013. PubMed PMID: 26083920; PubMed Central PMCID: PMC4472079.