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

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 biolo...

Full description

Saved in:
Bibliographic Details
Published inNature methods Vol. 16; no. 6; pp. 501 - 504
Main Authors Yang, Bin, Chen, Xingye, Wang, Yina, Feng, Siyu, Pessino, Veronica, Stuurman, Nico, Cho, Nathan H, Cheng, Karen W, Lord, Samuel J, Xu, Linfeng, Xie, Dan, Mullins, R Dyche, Leonetti, Manuel D, Huang, Bo
Format Journal Article
LanguageEnglish
Published United States Nature Publishing Group 01.06.2019
Subjects
Animals
Aperture
Bioengineering
Biophysics
Cameras
Cell lines
Design
Drosophila - metabolism
Efficiency
Fluorescence
Fluorescence microscopy
HEK293 Cells
Humans
Illumination
Image Processing, Computer-Assisted - methods
Imaging
Imaging, Three-Dimensional - methods
Light
Lighting - instrumentation
Localization
Methods
Microscopy
Microscopy, Fluorescence - methods
Molecular Imaging - methods
Objectives
Recording
Sample holders
Single-Cell Analysis - methods
Spatio-Temporal Analysis
Temporal resolution
Online AccessGet full text

Cover

More Information
Summary: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.
Bibliography:ObjectType-Article-1
SourceType-Scholarly Journals-1
ObjectType-Feature-2
content type line 23
B.Y. conceived, designed and built the microscope and performed the simulations and the experiments. B.Y., Y.W., S.F., V.P., N.C. and K.C. prepared the cell samples. B.Y. and N.S. implemented the Micro-Manager software for device control and wrote a custom script for fast data acquisition. B.Y. and X.C. built the system with a Ti-E microscope stand, automated the microscope and performed the long-term parallel imaging. L.X. drew the 3D solid model. B.Y., X.C., Y.W., D.X., S.J.L. and B.H. analyzed the data. R.D.M. supervised K.C. in sample preparation. M.D.L. supervised N.C. in sample preparation. B.H supervised the project. B.Y and B.H wrote the manuscript with input from all authors.
Author contributions
ISSN:1548-7091
1548-7105
DOI:10.1038/s41592-019-0401-3