SunRISE: long-term imaging of individual mRNA molecules in living cells

Mapping Intimacies ◽

10.1101/2020.10.05.326660 ◽

2020 ◽

Author(s):

Yue Guo ◽

Robin E. C. Lee

Keyword(s):

Phase Separation ◽

Single Cell ◽

Cell Imaging ◽

Living Cells ◽

Mrna Detection ◽

Central Dogma ◽

Single Cell Imaging ◽

Cell Fixation ◽

Limited Sensitivity

AbstractSingle-cell imaging of individual mRNAs has revealed core mechanisms of the central dogma. However, most approaches require cell fixation or have limited sensitivity for live-cell applications. Here, we describe SunRISE (SunTag-based Reporter for Imaging Signal Enriched mRNA), a computationally and experimentally optimized approach for unambiguous single-mRNA detection in living cells. We demonstrate SunRISE with long-term epifluorescence imaging, using translational stress to track mRNA phase separation and recovery from cytosolic droplets.

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Long-term single-cell imaging of mammalian stem cells

Nature Methods ◽

10.1038/nmeth.1577 ◽

2011 ◽

Vol 8 (S4) ◽

pp. S30-S35 ◽

Cited By ~ 121

Author(s):

Timm Schroeder

Keyword(s):

Stem Cells ◽

Single Cell ◽

Cell Imaging ◽

Single Cell Imaging

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Faculty Opinions recommendation of Microglia turnover with aging and in an Alzheimer's model via long-term in vivo single-cell imaging.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.729490031.793538321 ◽

2017 ◽

Author(s):

David Attwell ◽

Christian Madry

Keyword(s):

Single Cell ◽

Cell Imaging ◽

Single Cell Imaging

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Microglia turnover with aging and in an Alzheimer's model via long-term in vivo single-cell imaging

Nature Neuroscience ◽

10.1038/nn.4631 ◽

2017 ◽

Vol 20 (10) ◽

pp. 1371-1376 ◽

Cited By ~ 123

Author(s):

Petra Füger ◽

Jasmin K Hefendehl ◽

Karthik Veeraraghavalu ◽

Ann-Christin Wendeln ◽

Christine Schlosser ◽

...

Keyword(s):

Single Cell ◽

Cell Imaging ◽

Single Cell Imaging

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Understanding cell fate control by continuous single-cell quantification

Blood ◽

10.1182/blood-2018-09-835397 ◽

2019 ◽

Vol 133 (13) ◽

pp. 1406-1414 ◽

Cited By ~ 6

Author(s):

Dirk Loeffler ◽

Timm Schroeder

Keyword(s):

Single Cell ◽

Cell Fate ◽

Cell Imaging ◽

Population Based ◽

Molecular Processes ◽

Cell Fate Decisions ◽

Single Cell Imaging

Abstract Cells and the molecular processes underlying their behavior are highly dynamic. Understanding these dynamic biological processes requires noninvasive continuous quantitative single-cell observations, instead of population-based average or single-cell snapshot analysis. Ideally, single-cell dynamics are measured long-term in vivo; however, despite progress in recent years, technical limitations still prevent such studies. On the other hand, in vitro studies have proven to be useful for answering long-standing questions. Although technically still demanding, long-term single-cell imaging and tracking in vitro have become valuable tools to elucidate dynamic molecular processes and mechanisms, especially in rare and heterogeneous populations. Here, we review how continuous quantitative single-cell imaging of hematopoietic cells has been used to solve decades-long controversies. Because aberrant cell fate decisions are at the heart of tissue degeneration and disease, we argue that studying their molecular dynamics using quantitative single-cell imaging will also improve our understanding of these processes and lead to new strategies for therapies.

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