A portable microfluidic fluorescence spectrometer device for γ-H2AX-based biological dosimetry

Following a radiological incident the rapid identification of those individuals exposed to critically high radiation doses is important for initial triage and medical treatment. It has been previously demonstrated that scoring of radiation-induced foci of the phosphorylated histone γ-H2AX, which form at the sites of DNA double-strand breaks, may be used to determine radiation exposure levels from blood samples. Although faster than the ‘gold standard’ dicentric assay, foci scoring is still impractical in a field situation where large numbers of people may need to be screened. To deal with such a situation, an inexpensive portable device with high throughput capacity is desirable. Here we describe a portable microfluidic fluorescence spectrometer device which passes a suspension of γ-H2AX immunofluorescence-stained lymphocytes through a focused 488 nm laser beam in a microfluidic chamber and records emission spectra over the range 495–725 nm. The recorded emission spectra are spectrally unmixed into their constituent parts from which radiation exposure levels are determined. Proof of principle is demonstrated using cultured lymphoblastoid cells, exposed to X-ray doses between 0 and 8 Gy. With the current prototype setup it takes approximately 6 min to acquire and analyse 10,000 spectra. Further effort is required to fully develop this approach into a portable triage tool that could be used to help classify people into appropriate treatment categories based on radiation exposure levels.

[1]  Zbigniew Darzynkiewicz,et al.  Cytometric assessment of DNA damage in relation to cell cycle phase and apoptosis , 2005, Cell proliferation.

[2]  K. Nakachi,et al.  Short‐term culture and γH2AX flow cytometry determine differences in individual radiosensitivity in human peripheral T lymphocytes , 2007, Environmental and molecular mutagenesis.

[3]  Michael Uder,et al.  In vivo formation and repair of DNA double-strand breaks after computed tomography examinations. , 2005, Proceedings of the National Academy of Sciences of the United States of America.

[4]  S. Gasser,et al.  Crosstalk between histone modifications during the DNA damage response. , 2009, Trends in cell biology.

[5]  William F. Blakely,et al.  The Use of Gamma-H2AX as a Biodosimeter for Total-Body Radiation Exposure in Non-Human Primates , 2010, PloS one.

[6]  Jian Zhang,et al.  THE RABIT: A RAPID AUTOMATED BIODOSIMETRY TOOL FOR RADIOLOGICAL TRIAGE , 2010, Health physics.

[7]  E. Rogakou,et al.  Megabase Chromatin Domains Involved in DNA Double-Strand Breaks in Vivo , 1999, The Journal of cell biology.

[8]  Ruth C Wilkins,et al.  The response of gamma-H2AX in human lymphocytes and lymphocytes subsets measured in whole blood cultures , 2009, International journal of radiation biology.

[9]  Borivoj Vojnovic,et al.  Applications of cost-effective spectral imaging microscopy in cancer research , 2003 .

[10]  Vicky Goh,et al.  Leukocyte DNA damage after multi-detector row CT: a quantitative biomarker of low-level radiation exposure. , 2007, Radiology.

[11]  Kai Rothkamm,et al.  Evidence for a lack of DNA double-strand break repair in human cells exposed to very low x-ray doses , 2003, Proceedings of the National Academy of Sciences of the United States of America.

[12]  E. Rogakou,et al.  Quantitative Detection of 125IdU-Induced DNA Double-Strand Breaks with γ-H2AX Antibody , 2002 .

[13]  O. Hammarsten,et al.  An optimized method for detecting gamma-H2AX in blood cells reveals a significant interindividual variation in the gamma-H2AX response among humans , 2007, Nucleic acids research.

[14]  E. Rogakou,et al.  Quantitative detection of (125)IdU-induced DNA double-strand breaks with gamma-H2AX antibody. , 2002, Radiation research.

[15]  Kai Rothkamm,et al.  Candidate protein biomarkers as rapid indicators of radiation exposure , 2011 .