Counting Growth Factors in Single Cells with Infrared Quantum Dots to Measure Discrete Stimulation Distributions

Overview

Journal Nat Commun

Specialty Biology

Date 2019 Feb 24

PMID 30796217

Citations 9

Authors

Phuong Le

Sung Jun Lim

Brian C Baculis

Hee Jung Chung

Kristopher A Kilian

Andrew M Smith

Affiliations

Soon will be listed here.

Abstract

The distribution of single-cell properties across a population of cells can be measured using diverse tools, but no technology directly quantifies the biochemical stimulation events regulating these properties. Here we report digital counting of growth factors in single cells using fluorescent quantum dots and calibrated three-dimensional deconvolution microscopy (QDC-3DM) to reveal physiologically relevant cell stimulation distributions. We calibrate the fluorescence intensities of individual compact quantum dots labeled with epidermal growth factor (EGF) and demonstrate the necessity of near-infrared emission to overcome intrinsic cellular autofluoresence at the single-molecule level. When applied to human triple-negative breast cancer cells, we observe proportionality between stimulation and both receptor internalization and inhibitor response, reflecting stimulation heterogeneity contributions to intrinsic variability. We anticipate that QDC-3DM can be applied to analyze any peptidic ligand to reveal single-cell correlations between external stimulation and phenotypic variability, cell fate, and drug response.

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