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Single Cell Transfection in Chick Embryos

Single Cell Transfection in Chick Embryos

Published on: September 25, 2010

Counting low-copy number proteins in a single cell.

Bo Huang¹, Hongkai Wu, Devaki Bhaya

¹Department of Chemistry, Stanford University, Stanford, CA 94305-5080, USA.

Science (New York, N.Y.)

|January 6, 2007

Summary

This study introduces a microfluidic device for single-cell protein analysis. It enables precise quantification of protein levels in individual cells, revealing differences in cellular protein expression.

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Area of Science:

Biotechnology
Analytical Chemistry
Cell Biology

Background:

Single-cell analysis is crucial for understanding cellular heterogeneity.
Existing methods often lack the sensitivity or throughput for comprehensive single-cell protein quantification.

Purpose of the Study:

To develop and validate a microfluidic device for comprehensive single-cell protein analysis.
To enable high-efficiency single-molecule fluorescence counting for protein quantification.

Main Methods:

Design and implementation of a microfluidic device for cell manipulation, lysis, labeling, separation, and quantification.
Utilizing generic fluorescent-antibody binding for protein labeling.
Employing cylindrical optics for high-efficiency (approx. 60%) molecule counting in micrometer channels.

Main Results:

Successful quantification of beta2 adrenergic receptors in insect (SF9) cells.
Analysis of phycobiliprotein content in individual cyanobacterial (Synechococcus sp. PCC 7942) cells.
Observed significant differences in specific complex levels in cyanobacteria grown under nitrogen-depleted conditions.

Conclusions:

The developed microfluidic device offers a powerful platform for single-cell protein analysis.
The device demonstrates high efficiency in molecule counting, enabling sensitive detection.
The findings highlight the utility of the device for studying cellular responses to environmental changes.