Epi-illumination SPIM for volumetric imaging with high spatial-temporal resolution

Bin Yang; Xingye Chen; Yina Wang; Siyu Feng; Veronica Pessino; Nico Stuurman; Nathan H Cho; Karen W Cheng; Samuel J Lord; Linfeng Xu; Dan Xie; R Dyche Mullins; Manuel D Leonetti; Bo Huang

doi:10.1038/s41592-019-0401-3

Epi-illumination SPIM for volumetric imaging with high spatial-temporal resolution

Nat Methods. 2019 Jun;16(6):501-504. doi: 10.1038/s41592-019-0401-3. Epub 2019 May 6.

Authors

Bin Yang¹, Xingye Chen^{1

2}, Yina Wang¹, Siyu Feng³, Veronica Pessino^{1

4}, Nico Stuurman^{5

6}, Nathan H Cho⁷, Karen W Cheng⁵, Samuel J Lord^{5

6}, Linfeng Xu⁸, Dan Xie¹, R Dyche Mullins^{5

6}, Manuel D Leonetti⁷, Bo Huang^{9

10

11}

Affiliations

¹ Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA, USA.
² Department of Automation, Tsinghua University, Beijing, China.
³ The UC Berkeley-UCSF Graduate Program in Bioengineering, University of California, San Francisco, San Francisco, CA, USA.
⁴ Biophysics Graduate Program, University of California, San Francisco, San Francisco, CA, USA.
⁵ Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA, USA.
⁶ Howard Hughes Medical Institute, San Francisco, CA, USA.
⁷ Chan Zuckerberg Biohub, San Francisco, CA, USA.
⁸ Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA, USA.
⁹ Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA, USA. bo.huang@ucsf.edu.
¹⁰ Chan Zuckerberg Biohub, San Francisco, CA, USA. bo.huang@ucsf.edu.
¹¹ Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA, USA. bo.huang@ucsf.edu.

Abstract

We designed an epi-illumination SPIM system that uses a single objective and has a sample interface identical to that of an inverted fluorescence microscope with no additional reflection elements. It achieves subcellular resolution and single-molecule sensitivity, and is compatible with common biological sample holders, including multi-well plates. We demonstrated multicolor fast volumetric imaging, single-molecule localization microscopy, parallel imaging of 16 cell lines and parallel recording of cellular responses to perturbations.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Animals
Drosophila / metabolism*
HEK293 Cells
Humans
Image Processing, Computer-Assisted / methods*
Imaging, Three-Dimensional / methods*
Lighting / instrumentation*
Microscopy, Fluorescence / methods*
Molecular Imaging / methods*
Single-Cell Analysis / methods*
Spatio-Temporal Analysis

Abstract

Publication types

MeSH terms

Grants and funding