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...
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Published in | Nature methods Vol. 16; no. 6; pp. 501 - 504 |
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Main Authors | , , , , , , , , , , , , , |
Format | Journal Article |
Language | English |
Published |
United States
Nature Publishing Group
01.06.2019
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Subjects | |
Online Access | Get full text |
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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. |
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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 |