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DNA Microarrays

DNA Microarrays

Microarrays are high-throughput and relatively inexpensive assays that can be automated to analyze large quantities of data at a time. They are used in genome-wide studies to compare gene or protein expression under two varied conditions, such as healthy and diseased states. Microarrays consist of glass or silica slides on which probe molecules are covalently attached through surface functionalization. Most commonly, the slides are prepared through the chemisorption of silanes to silica...

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Real Time RT-PCR

Real-time reverse transcription-polymerase chain reaction, or Real-time RT-PCR, is an analytical tool used to determine the expression level of target genes. The method involves converting mRNA to complementary DNA with the help of an enzyme known as reverse transcriptase, followed by the PCR amplification of the cDNA. These two processes can be performed simultaneously in a single tube or separately as a two-step reaction.
The real-time quantification of the number of amplified products is...

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Related Experiment Video

Updated: May 1, 2026

Counting Proteins in Single Cells with Addressable Droplet Microarrays

Counting Proteins in Single Cells with Addressable Droplet Microarrays

Published on: July 6, 2018

Absolute quantification of protein copy number using a single-molecule-sensitive microarray.

Edward Burgin¹, Ali Salehi-Reyhani, Michael Barclay

¹Single Cell Proteomics Project, Institute of Chemical Biology, Imperial College London, London, SW7 2AZ, UK. d.klug@imperial.ac.uk.

|March 29, 2014

Summary

This summary is machine-generated.

This study introduces a microfluidic microarray for precise protein quantification using single molecule detection. The developed assays enable calibration-free measurement of protein copy numbers in solution and at the single-cell level.

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Area of Science:

Biotechnology
Analytical Chemistry
Molecular Biology

Background:

Accurate protein quantification is crucial for biological research and diagnostics.
Existing methods often require extensive calibration or lack sensitivity for low-abundance proteins.

Purpose of the Study:

To develop and validate a microfluidic microarray system for absolute protein quantification.
To demonstrate calibration-free detection protocols for enhanced usability.
To determine protein copy numbers at the single-cell level.

Main Methods:

Utilized a microfluidic microarray platform with single molecule detection capabilities.
Developed and tested two protein detection assays: one for EGFP protein and one antibody sandwich assay for p53.
Validated the system for absolute quantification of protein copy number in solution and within cancer cell lines.

Main Results:

Achieved a limit of detection of <30 EGFP proteins and ~21 p53 proteins.
Demonstrated a dynamic range of >5 orders of magnitude for EGFP and >3 orders of magnitude for p53.
Successfully applied the protocols for retrospective calibration to determine absolute protein copy number at the single-cell level in human cancer cell lines.

Conclusions:

The microfluidic microarray system enables accurate and sensitive absolute protein quantification.
Calibration-free protocols enhance the practicality and accessibility of the technology.
This method provides a powerful tool for single-cell protein analysis in cancer research.