Abstract

Fluorescence in situ hybridization (FISH) is the primary technology used to image and count mRNA in single cells, but applications of the technique are limited by photophysical shortcomings of organic dyes. Inorganic quantum dots (QDs) can overcome these problems but years of development have not yielded viable QD-FISH probes. Here we report that macromolecular size thresholds limit mRNA labeling in cells, and that a new generation of compact QDs produces accurate mRNA counts. Compared with dyes, compact QD probes provide exceptional photostability and more robust transcript quantification due to enhanced brightness. New spectrally engineered QDs also allow quantification of multiple distinct mRNA transcripts at the single-molecule level in individual cells. We expect that QD-FISH will particularly benefit high-resolution gene expression studies in three dimensional biological specimens for which quantification and multiplexing are major challenges.

Details

Title
Enhanced mRNA FISH with compact quantum dots
Author
Liu, Yang 1 ; Le, Phuong 1 ; Lim, Sung Jun 2 ; Ma, Liang 3   VIAFID ORCID Logo  ; Sarkar, Suresh 1 ; Han, Zhiyuan 3 ; Murphy, Stephen J 4   VIAFID ORCID Logo  ; Kosari, Farhad 4 ; Vasmatzis, George 4 ; Cheville, John C 5 ; Smith, Andrew M 6   VIAFID ORCID Logo 

1  Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, Urbana, IL, USA 
2  Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Intelligent Devices and Systems Research Group, DGIST, Hyeonpung, Daegu, Republic of Korea 
3  Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA 
4  Biomarker Discovery Program, Center of Individualized Medicine, Mayo Clinic, Rochester, MN, USA 
5  Biomarker Discovery Program, Center of Individualized Medicine, Mayo Clinic, Rochester, MN, USA; Department of Laboratory Medicine and Pathology, Mayo Clinic, Rochester, MN, USA 
6  Department of Bioengineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Micro and Nanotechnology Laboratory, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Department of Materials Science and Engineering, University of Illinois at Urbana-Champaign, Urbana, IL, USA; Carle Illinois College of Medicine, Urbana, IL, USA 
Pages
1-8
Publication year
2018
Publication date
Oct 2018
Publisher
Nature Publishing Group
e-ISSN
20411723
Source type
Scholarly Journal
Language of publication
English
DOI
https://doi.org/10.1038/s41467-018-06740-x
ProQuest document ID
2125648361
Copyright
© 2018. This work is published under http://creativecommons.org/licenses/by/4.0/ (the “License”). Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.