Observing the cell in its native state: Imaging subcellular dynamics in multicellular organisms

Tsung Li Liu, Srigokul Upadhyayula, Daniel E. Milkie, Ved Singh, Kai Wang, Ian A. Swinburne, Kishore R. Mosaliganti, Zach M. Collins, Tom W. Hiscock, Jamien Shea, Abraham Q. Kohrman, Taylor N. Medwig, Daphne Dambournet, Ryan Forster, Brian Cunniff, Yuan Ruan, Hanako Yashiro, Steffen Scholpp, Elliot M. Meyerowitz, Dirk HockemeyerDavid G. Drubin, Benjamin L. Martin, David Q. Matus, Minoru Koyama, Sean G. Megason, Tom Kirchhausen, Eric Betzig*

*Corresponding author for this work

Research output: Contribution to journalArticlepeer-review

372 Citations (Scopus)


True physiological imaging of subcellular dynamics requires studying cells within their parent organisms, where all the environmental cues that drive gene expression, and hence the phenotypes that we actually observe, are present. A complete understanding also requires volumetric imaging of the cell and its surroundings at high spatiotemporal resolution, without inducing undue stress on either. We combined lattice light-sheet microscopy with adaptive optics to achieve, across large multicellular volumes, noninvasive aberration-free imaging of subcellular processes, including endocytosis, organelle remodeling during mitosis, and the migration of axons, immune cells, and metastatic cancer cells in vivo. The technology reveals the phenotypic diversity within cells across different organisms and developmental stages and may offer insights into how cells harness their intrinsic variability to adapt to different physiological environments.

Original languageEnglish
Article numbereaaq1392
Number of pages15
Issue number6386
Early online date20 Apr 2018
Publication statusPublished - 20 Apr 2018
Externally publishedYes

Bibliographical note

Acknowledgments: We thank the Shared Resource teams at the Janelia Research Campus for their skill and dedication in specimen handling and preparation, K. Schaefer for the preparation of cancer cells for the xenograph experiments, and the Instrument Design and Fabrication team for their manufacturing expertise. We also gratefully acknowledge the support of the Janelia Visitor Program. S.U. thanks H. Elliott, D. Richmond, and R. Gao for discussions and acknowledges the MATLAB code repository received from the Computational Image Analysis Workshop supported by NIH grant GM103792.

Funding: T.-L.L., D.E.M., V.S., J.S., M.K., E.M.M., and E.B. are funded by the Howard Hughes Medical Institute (HHMI). T.K. and S.U. are funded by grants from Biogen, Ionis Pharmaceuticals, and NIH grant R01GM075252 (to T.K.). S.U. is a Fellow at the Image and Data Analysis core at Harvard Medical School. S.S. is funded by a Living Systems Institute start-up grant, University of Exeter. K.R.M., I.A.S., Z.M.C., T.W.H., and S.G.M. were supported by NIH grant R01DC015478. D.Q.M. is funded by the NIH (5R00CA154870-05 and 1R01GM121597-01). D.Q.M. and B.L.M. are funded by the Carol M. Baldwin Foundation and are Damon Runyon-Rachleff Innovators supported (in part) by the Damon Runyon Cancer Research Foundation (DRR-47-17). B.L.M. is also funded by the NSF (IOS1452928). D.H. is a Pew-Stewart Scholar for Cancer Research supported by the Pew Charitable Trusts and NIH grant R01CA196884. D.D. was supported by a Human Frontier Science Program Fellowship. D.G.D. was supported by NIH grant R35GM118149.

Author contributions: E.B. supervised the project and wrote the manuscript with input from all coauthors. T.-L.L. built the microscope with input from E.B., D.E.M., and K.W. and performed all microscope characterization experiments. D.E.M. created the instrument control software. T.-L.L., V.S., and S.U. acquired all biological data with coauthors. D.D., D.G.D., R.F., and D.H. provided organoids and led related experiments. T.K., S.U., B.C., and S.S. provided transgenic zebrafish and AP2 cells and led related clathrin experiments. Z.M.C., T.W.H., and S.G.M. created the mCardinal-PM transgenic zebrafish. T.K., S.G.M., S.U., and I.A.S. provided zebrafish and led related organelle dynamics and in vivo immune cell experiments. J.S. and M.K. created the Autobow zebrafish, and M.K. led related experiments. A.Q.K., T.N.M., and D.Q.M. provided C. elegans specimens and led related experiments. B.L.M. provided MDA-MB-231 cells and vasculature-labeled zebrafish and led related experiments with D.Q.M. Y.R., H.Y., and E.M.M. provided Arabidopsis specimens and led related experiments. S.U., K.R.M., T.-L.L., and V.S. processed all image data. S.U. performed quantitative analysis of all image data. S.U., T.-L.L., and E.B. produced all figures and movies. Competing interests: Portions of the technology described herein are covered by U.S. Patent 7,894,136 issued to E.B., assigned to Lattice Light of Ashburn, VA, and licensed to Carl Zeiss Microscopy; U.S. Patents 8,711,211 and 9,477,074 issued to E.B., assigned to HHMI, and licensed to Carl Zeiss Microscopy; U.S. Patent application 13/844,405 filed by E.B. and K.W. and assigned to HHMI; and U.S. Patent 9,500,846 issued to E.B. and K.W. and assigned to HHMI.

Data and materials availability: All data needed to evaluate the conclusions in the paper are present in the paper or the supplementary materials. Documentation for construction of a lattice light-sheet microscope can be obtained by execution of a research license agreement with HHMI.


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