Enhanced MRNA FISH with Compact Quantum Dots

Overview

Journal Nat Commun

Specialty Biology

Date 2018 Oct 28

PMID 30367061

Citations 16

Authors

Yang Liu

Phuong Le

Sung Jun Lim

Liang Ma

Suresh Sarkar

Zhiyuan Han

Stephen J Murphy

Farhad Kosari

George Vasmatzis

John C Cheville

Andrew M Smith

Affiliations

Soon will be listed here.

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.

Citing Articles

Visualization of Individual RNA Molecules by Proximity Ligation-Based Chromogenic In Situ Hybridization Assay.

Xia X, Jiang M, Lin C, Ke R Methods Mol Biol. 2024; 2822:143-156.

PMID: 38907917 DOI: 10.1007/978-1-0716-3918-4_11.

Quantum Dot-Fab' Conjugates as Compact Immunolabels for Microtubule Imaging and Cell Classification.

Kolossov V, Kanakaraju K, Sarkar S, Arogundade O, Kuo C, Mara N ACS Nano. 2024; 18(23):15084-15095.

PMID: 38815170 PMC: 11262708. DOI: 10.1021/acsnano.4c02215.

Single-Molecule Fluorescent In Situ Hybridization (smFISH) for RNA Detection in Bacteria.

Mattioli C, Avraham R Methods Mol Biol. 2024; 2784:3-23.

PMID: 38502475 DOI: 10.1007/978-1-0716-3766-1_1.

Digital and Absolute Assays for Low Abundance Molecular Biomarkers.

Kuo C, Smith A Acc Chem Res. 2023; 56(9):1031-1042.

PMID: 37068158 PMC: 10163903. DOI: 10.1021/acs.accounts.3c00030.

Structural Design of Multidentate Copolymers as Compact Quantum Dot Coatings for Live-Cell Single-Particle Imaging.

Han Z, Vaidya R, Arogundade O, Ma L, Zahid M, Sarkar S Chem Mater. 2023; 34(10):4621-4632.

PMID: 36968145 PMC: 10038122. DOI: 10.1021/acs.chemmater.2c00498.

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