Individual and Population Quantitative Analyses of Calcium Flux in T-Cells Activated on Functionalized Material Surfaces
Susan N. Christo A B , Ghafar.T. Sarvestani C , Stefani S. Griesser D , Bryan R. Coad D E , Hans J. Griesser D , Krasimir Vasilev E , Michael P. Brown A F G , Kerrilyn R. Diener A H and John D. Hayball A B H IA Experimental Therapeutics Laboratory, Hanson Institute, Royal Adelaide Hospital, PO Box 14 Rundle Mall, Adelaide, SA 5000, Australia.
B Sansom Institute, University of South Australia, PO Box 2471, Adelaide, SA 5001, Australia.
C Detmold Imaging Core Facility, SA Pathology, PO Box 14, Rundle Mall, Adelaide, SA 5000, Australia.
D Ian Wark Research Institute, University of South Australia, Mawson Lakes Campus, Mawson Lakes, Adelaide, SA 5095, Australia.
E Mawson Institute, University of South Australia, PO Box 2471, Adelaide, SA 5001, Australia.
F Cancer Clinical Trials Unit, Royal Adelaide Hospital, North Terrace, Adelaide, SA 5000, Australia.
G School of Medicine, University of Adelaide, Adelaide, SA 5005, Australia. SA 5005, Australia.
H School of Paediatrics and Reproductive Health, University of Adelaide, Adelaide, SA 5005, Australia.
I Corresponding author. School of Pharmacy and Medical Science, University of South Australia, Frome Road, Adelaide, South Australia, Australia, 5000. Email: john.hayball@unisa.edu.au
Australian Journal of Chemistry 65(1) 45-49 https://doi.org/10.1071/CH11311
Submitted: 25 July 2011 Accepted: 16 October 2011 Published: 23 November 2011
Abstract
We have developed a novel method for activating T-cells on material surfaces that enable individual and population-based analyses of intracellular calcium flux, as a quantitative measure of T-cell receptor engagement. Functionalized material surfaces were created using a plasma-polymerized foundation layer to immobilize stimulatory T-cell ligands, which could induce T-cell receptor-dependent calcium flux in naive T-cells. Real-time confocal microscopic detection and quantification of calcium flux using paired fluorescent ratiometric probes facilitated the tracking and analysis of response profiles of individual T-cells, as well as population analyses using a combination of individual T-cell events. This type of combined analysis cannot be achieved using traditional population-based flow cytometric approaches, and thus provides a logical step towards developing the capacity to assess the magnitude and quality of inherently heterogeneous effector T-cell responses to antigenic challenge.
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