Work

Quantitative analysis of protein translocations by microfluidic total internal reflection fluorescence flow cytometry

Year: 2010

Type: article

Abstract: Protein translocation, or the change in a protein's location between different subcellular compartments, is a critical process by which intracellular proteins carry out their cellular functions. Aberrant translocation events contribute to various diseases ranging from metabolic disorders to cancer. In this study, we demonstrate the use of a newly developed single-cell tool, microfluidic total internal reflection fluorescence flow cytometry (TIRF-FC), for detecting both cytosol to plasma membrane and cytosol to nucleus translocations using the tyrosine kinase Syk and the transcription factor NF-κB as models. This technique detects fluorescent molecules at the plasma membrane and in the membrane-proximal cytosol in single cells. We were able to record quantitatively changes in the fluorescence density in the evanescent field associated with these translocation processes for large cell populations with single cell resolution. We envision that TIRF-FC will provide a new approach to explore the molecular biology and clinical relevance of protein translocations.

Source: Lab on a Chip

DOI: 10.1039/c0lc00131g

Authors: Jun Wang, Bei Fei, Robert L. Geahlen, Chang Lu

Institutions: Purdue University West Lafayette, Virginia Tech

Language: English

Topic: Advanced Fluorescence Microscopy Techniques

Subfield: Biophysics

Field: Biochemistry, Genetics and Molecular Biology

Domain: Life Sciences

SDG: Good health and well-being

Open Access status: green

Funders: National Science Foundation, National Institutes of Health, National Cancer Institute, Division of Chemical, Bioengineering, Environmental, and Transport Systems