Abstract
Image processing methods and techniques for high-throughput quantification of dielectrophoretic (DEP) collections onto planar castellated electrode arrays are developed and evaluated. Fluorescence-based dielectrophoretic spectroscopy is an important tool for laboratory investigations of AC electrokinetic properties of nanoparticles. This paper details new, first principle, theoretical and experimental developments of geometric feature recognition techniques that enable quantification of positive dielectrophoretic (pDEP) nanoparticle collections onto castellated arrays. As an alternative to the geometric-based method, novel statistical methods that do not require any information about array features, are also developed using the quantile and standard deviation functions. Data from pDEP collection and release experiments using 200 nm diameter latex nanospheres demonstrates that pDEP quantification using the statistic-based methods yields quantitatively similar results to the geometric-based method. The development of geometric- and statistic-based quantification methods enables high-throughput, supervisor-free image processing tools critical for dielectrophoretic spectroscopy and automated DEP technology development.
Highlights
Nanoparticle technologies have been developing rapidly over the last decade with many practical applications in nano-medicine
Dual-cycle pulsed positive dielectrophoretic (pDEP) collection, a technique recently developed for determining the dielectric properties of nanoparticles,[4,5,6] was used as a method for an experimental example with amplitude modulated (AM) control and probe frequencies, 0.7 MHz and 3.0 MHz, respectively; and we demonstrated agreement between the geometric method and the statistic-based quantile and standard deviation methods to within 15%
Two important advancements in image quantification tools and methods for DEP-driven nanoparticle movement using planar castellated electrode arrays have been reported in detail
Summary
Nanoparticle technologies have been developing rapidly over the last decade with many practical applications in nano-medicine (e.g. diagnostics and drug delivery). Applications of pDEP using planar electrode arrays include measurements of collection rates for characterising cells and their constituents, e.g. DNA, RNA, viruses, and colloidal bioparticles.[7,8,9,10,11,12,13,14,15,16,17] Recently, our groups demonstrated continuously pulsed pDEP as a robust and fast alternative to crossover measurements[10,13,18,19,20] by quantifying initial collection rates where the nanoparticle capture rate is approximately proportional the DEP force.[7,8,9,10,11,12,14,15,16,17] The collection rates were, in turn, fitted to the RF dependent polarizability modelled by the real part of the Clausius-Mossotti function.[4,5,6] DEP video recorded fluorescence microscope dielectrophoretic spectroscopy is becoming an important tool for laboratory investigations of AC electrokinetic nanoparticle behaviour and could advance novel investigations, e.g. probing biological and chemical interactions at the nano-scale.[21,22,23,24]. Quantitative comparisons are made between the two methods of DEP collection experiments using 200 nm latex particles
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