DMMC Course: TECHNIQUES & STRATEGIES IN MOLECULAR MEDICINE

1130-1215 Thursday 13 December 2007, Panoz institute, LTEE2, TCD

High Content Analysis of nanoparticle/cell interactions

Dr Yuri Volkov (Institute of Molecular Medicine, TCD)

Fluorescent organic tags have represented one of the major tools in the arsenal of researchers working in the biomedical sciences for more that two decades. The progress in development of new fast and efficient research and diagnostic methods is largely dependent on the availability of fluorescent probes with desired cell receptor- and organelle specificity and optimised experimental protocols for their utilization.

A unique opportunity to generate a wide spectrum of such probes suitable for applications in living cells is offered by semiconductor quantum dots (QDs). As fluorescent probes QDs have several advantages over organic dyes, including wide absorption profiles, tunable emission spectra, and superior photostability. QDs have been shown to readily distribute across animal cells, tissues and organs. Today, QDs with different physico-chemical properties and functionalities are readily available worldwide. However, further exploitation of QDs in biomedical studies has been hindered by the absence of adequate technological platforms capable of performing multi-parametric quantitative analysis of individual responses in specific cell types.

Recent years have witnessed a rapid progress in the development of novel methods permitting high-resolution visualisation of cell receptor dynamics and intracellular biochemical processes utilizing fluorescent probes. Among these, high content screening (HCS) technology allows to perform analysis of molecular interactions in individual cells and their populations at the sub-cellular level under physiological conditions. This technology not only facilitates development of a better understanding of the true functionality of target molecules in the living cells, but it can also promote designing of highly informative screens for novel therapeutic drugs, including inhibitors at small molecule and gene therapy level applicable in inflammation and cancer.

Here we will discuss the data demonstrating the influence of such factors as QDs size, charge and selective functionalisation on their membrane and subcellular localisation specificity and present an overview of advantages and hurdles on the way of merging nanotechnology and high content cell analysis.