Format

Send to

Choose Destination
See comment in PubMed Commons below
BMC Res Notes. 2011 Sep 9;4:340. doi: 10.1186/1756-0500-4-340.

The use of HaloTag-based technology in flow and laser scanning cytometry analysis of live and fixed cells.

Author information

1
Immune Disease Institute and Program in Cellular and Molecular Medicine, Children's Hospital Boston and Harvard Medical School, Boston, MA, USA. barteneva@idi.harvard.edu.

Abstract

BACKGROUND:

Combining the technologies of protein tag labeling and optical microscopy allows sensitive analysis of protein function in cells.

FINDINGS:

Here, we describe development of applications using protein tag technology (HaloTag (HT)-based) for flow and laser scanning cytometry (LSC). Cell lines, expressing recombinant surface β1-integrin-HT and HT-p65 fusion protein, and a CD4 T cell line (Jurkat) infected with human immunodeficiency virus type 1 (HIV-1) reporter virus expressing the unfused HT (HIV-1Lai-Halo), were stained with different HT ligands and successfully detected by flow cytometers equipped with 488 and 561 nm lasers as well as a laser scanning cytometer (equipped with 488 and 405 nm lasers) alone or combined with cell cycle and viability markers.

CONCLUSIONS:

Use of HT technology for cytometric applications has advantages over its use in microscopy as it allows for the statistical measurement of protein expression levels in individual cells within a heterogeneous cell population in combination with cell cycle analysis. Another advantage is the ability of the HaloTag to withstand long fixation and high concentration of fixative, which can be useful in research of infectious agents like HIV and/or mycobacteria.

PubMed Commons home

PubMed Commons

0 comments
How to join PubMed Commons

    Supplemental Content

    Full text links

    Icon for BioMed Central Icon for PubMed Central
    Loading ...
    Support Center