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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| 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. |
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| AbstractList | 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. Epi-illumination SPIM enables fast, volumetric, high-resolution, subcellular imaging of any sample compatible with a standard inverted fluorescence microscope. We designed an epi-illumination SPIM system which utilizes a single objective and has an identical sample interface as 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 sixteen cell lines and parallel recording of cellular responses to perturbations. 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. |
| Audience | Academic |
| Author | Xu, Linfeng Xie, Dan Cho, Nathan H Feng, Siyu Cheng, Karen W Leonetti, Manuel D Chen, Xingye Wang, Yina Lord, Samuel J Huang, Bo Stuurman, Nico Yang, Bin Mullins, R Dyche Pessino, Veronica |
| AuthorAffiliation | 3. The UC Berkeley-UCSF Graduate Program in Bioengineering, San Francisco, CA 94143, USA 2. Department of Automation, Tsinghua University, Beijing 100084, China 4. Graduate Program of Biophysics, University of California, San Francisco, San Francisco, CA 94143, USA 5. Department of Cellular and Molecular Pharmacology, University of California, San Francisco, 600 16th Street, San Francisco, CA 94143, USA 7. Chan Zuckerberg Biohub, San Francisco, CA 94158, USA 1. Department of Pharmaceutical Chemistry, University of California in San Francisco, San Francisco, CA 94143, USA 6. Howard Hughes Medical Institute, San Francisco, CA 94143 USA 8. Department of Bioengineering and Therapeutic Sciences, University of California in San Francisco, San Francisco, CA 94143, USA 9. Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94143, USA |
| AuthorAffiliation_xml | – name: 2. Department of Automation, Tsinghua University, Beijing 100084, China – name: 8. Department of Bioengineering and Therapeutic Sciences, University of California in San Francisco, San Francisco, CA 94143, USA – name: 3. The UC Berkeley-UCSF Graduate Program in Bioengineering, San Francisco, CA 94143, USA – name: 1. Department of Pharmaceutical Chemistry, University of California in San Francisco, San Francisco, CA 94143, USA – name: 4. Graduate Program of Biophysics, University of California, San Francisco, San Francisco, CA 94143, USA – name: 6. Howard Hughes Medical Institute, San Francisco, CA 94143 USA – name: 5. Department of Cellular and Molecular Pharmacology, University of California, San Francisco, 600 16th Street, San Francisco, CA 94143, USA – name: 7. Chan Zuckerberg Biohub, San Francisco, CA 94158, USA – name: 9. Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA 94143, USA |
| Author_xml | – sequence: 1 givenname: Bin orcidid: 0000-0001-5138-4663 surname: Yang fullname: Yang, Bin organization: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA, USA – sequence: 2 givenname: Xingye surname: Chen fullname: Chen, Xingye organization: Department of Automation, Tsinghua University, Beijing, China – sequence: 3 givenname: Yina surname: Wang fullname: Wang, Yina organization: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA, USA – sequence: 4 givenname: Siyu surname: Feng fullname: Feng, Siyu organization: The UC Berkeley-UCSF Graduate Program in Bioengineering, University of California, San Francisco, San Francisco, CA, USA – sequence: 5 givenname: Veronica surname: Pessino fullname: Pessino, Veronica organization: Biophysics Graduate Program, University of California, San Francisco, San Francisco, CA, USA – sequence: 6 givenname: Nico orcidid: 0000-0002-6179-8613 surname: Stuurman fullname: Stuurman, Nico organization: Howard Hughes Medical Institute, San Francisco, CA, USA – sequence: 7 givenname: Nathan H orcidid: 0000-0002-0110-1343 surname: Cho fullname: Cho, Nathan H organization: Chan Zuckerberg Biohub, San Francisco, CA, USA – sequence: 8 givenname: Karen W surname: Cheng fullname: Cheng, Karen W organization: Department of Cellular and Molecular Pharmacology, University of California, San Francisco, San Francisco, CA, USA – sequence: 9 givenname: Samuel J orcidid: 0000-0002-2785-989X surname: Lord fullname: Lord, Samuel J organization: Howard Hughes Medical Institute, San Francisco, CA, USA – sequence: 10 givenname: Linfeng orcidid: 0000-0002-1358-2455 surname: Xu fullname: Xu, Linfeng organization: Department of Bioengineering and Therapeutic Sciences, University of California, San Francisco, San Francisco, CA, USA – sequence: 11 givenname: Dan surname: Xie fullname: Xie, Dan organization: Department of Pharmaceutical Chemistry, University of California, San Francisco, San Francisco, CA, USA – sequence: 12 givenname: R Dyche surname: Mullins fullname: Mullins, R Dyche organization: Howard Hughes Medical Institute, San Francisco, CA, USA – sequence: 13 givenname: Manuel D surname: Leonetti fullname: Leonetti, Manuel D organization: Chan Zuckerberg Biohub, San Francisco, CA, USA – sequence: 14 givenname: Bo orcidid: 0000-0003-1704-4141 surname: Huang fullname: Huang, Bo email: bo.huang@ucsf.edu, bo.huang@ucsf.edu, bo.huang@ucsf.edu organization: Department of Biochemistry and Biophysics, University of California, San Francisco, San Francisco, CA, USA. bo.huang@ucsf.edu |
| BackLink | https://www.ncbi.nlm.nih.gov/pubmed/31061492$$D View this record in MEDLINE/PubMed |
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| Copyright | COPYRIGHT 2019 Nature Publishing Group The Author(s), under exclusive licence to Springer Nature America, Inc. 2019. |
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| Notes | 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 |
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| Snippet | 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... We designed an epi-illumination SPIM system which utilizes a single objective and has an identical sample interface as an inverted fluorescence microscope with... |
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| SubjectTerms | 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 |
| Title | Epi-illumination SPIM for volumetric imaging with high spatial-temporal resolution |
| URI | https://www.ncbi.nlm.nih.gov/pubmed/31061492 https://www.proquest.com/docview/2232652509/abstract/ https://search.proquest.com/docview/2232133922 https://pubmed.ncbi.nlm.nih.gov/PMC6557432 |
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