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An image processing application for quantitative cross-correlative microscopy for large cell-populations: a gold nanoparticle radiosensitisation study

Published online by Cambridge University Press:  02 August 2017

Tyron Turnbull ,
Michael Douglass ,
Eva Bezak ,
Benjamin Thierry  and
Ivan Kempson
Tyron Turnbull
Affiliation:
Future Industries Institute, University of South Australia, Mawson Lakes Campus, Mawson Lakes, Adelaide, SA 5095, Australia
Michael Douglass
Affiliation:
Department of Medical Physics, Royal Adelaide Hospital, North Terrace, Adelaide, SA 5000, Australia School of Physical Sciences, University of Adelaide, Adelaide, SA 5005, Australia
Eva Bezak
Affiliation:
School of Physical Sciences, University of Adelaide, Adelaide, SA 5005, Australia International Centre for Allied Health Evidence and Sansom Insitute for Health Research, University of South Australia, City East Campus, North Terrace, Adelaide, SA 5001, Australia
Benjamin Thierry
Affiliation:
Future Industries Institute, University of South Australia, Mawson Lakes Campus, Mawson Lakes, Adelaide, SA 5095, Australia
Ivan Kempson*
Affiliation:
Future Industries Institute, University of South Australia, Mawson Lakes Campus, Mawson Lakes, Adelaide, SA 5095, Australia
*
a)Author to whom correspondence should be addressed. Electronic mail: [email protected]
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Abstract

A robust analysis script was developed in MATLAB for cross-correlative quantification of internalised gold nanoparticle (AuNP) uptake in a large number of individual cells with the corresponding number of DNA double-strand breaks (DSBs) in the same cells. The correlation of inorganic NP content with a biological marker at the single-cell level will aid in the elucidation of mechanisms of NP radiosensitisation. PC-3 cells were co-cultured with AuNPs and irradiated using an iridium-192 source. AuNP uptake was measured using synchrotron X-ray fluorescence (XRF) and DSBs imaged via confocal microscopy. MATLAB 2016a was used to develop a script to cross-correlate the two imaging modalities and quantify both DSBs and internalised AuNP content in the same cell. Various user-defined options written into the script give a high degree of versatility, which can account for a large number of variables in experimental parameters and data acquisition. The analysis procedure is flexible and robust, which gives consistent consideration to the wide spectrum of potential input image/data sets. Quantitative correlative microscopy was achieved with a custom MATLAB script used to correlate γH2AX foci (a marker of DNA DSBs) from confocal microscopy with AuNP content acquired using synchrotron XRF at the single-cell level. The script can be extended to a broad range of multi-modality imaging spectroscopies.

Keywords

radiation damage evaluation methodsimage processingX-ray fluorescence (XRF) systemsimaging spectroscopy
Type
Technical Articles
Information
Powder Diffraction , Volume 32 , Supplement S2 , December 2017 , pp. S33 - S37
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Copyright
Copyright © International Centre for Diffraction Data 2017 

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