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Proceedings Paper

Applications of quantitative time lapse holographic imaging to the development of complex pharmaceutical nano formulations
Author(s): Ed Luther; Livia Mendes; Jiayi Pan; Daniel Costa; Can Sarisozen; Vladimir Torchilin
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Paper Abstract

We rely on in vitro cellular cultures to evaluate the effects of the components of multifunctional nano-based formulations under development. We employ an incubator-adapted, label-free holographic imaging cytometer HoloMonitor M4® (Phase Holographic Imaging, Lund, Sweden) to obtain multi-day time-lapse sequences at 5- minute intervals. An automated stage allows hand-free acquisition of multiple fields of view.

Our system is based on the Mach-Zehnder interferometry principle to create interference patterns which are deconvolved to produce images of the optical thickness of the field of view. These images are automatically segmented resulting in a full complement of quantitative morphological features, such as optical volume, thickness, and area amongst many others. Precise XY cell locations and the time of acquisition are also recorded.

Visualization is best achieved by novel 4-Dimensional plots, where XY position is plotted overtime time (Z-directions) and cell-thickness is coded as color or gray scale brightness. Fundamental events of interest, i.e., cells undergoing mitosis or mitotic dysfunction, cell death, cell-to-cell interactions, motility are discernable. We use both 2D and 3D models of the tumor microenvironment.

We report our new analysis method to track feature changes over time based on a 4-sample version of the Kolmogorov-Smirnov test. Feature A is compared to Control A, and Feature B is compared to Control B to give a 2D probability plot of the feature changes over time. As a result, we efficiently obtain vectors quantifying feature changes over time in various sample conditions, i.e., changing compound concentrations or multi-compound combinations.

Paper Details

Date Published: 23 February 2018
PDF: 18 pages
Proc. SPIE 10503, Quantitative Phase Imaging IV, 105032G (23 February 2018); doi: 10.1117/12.2288504
Show Author Affiliations
Ed Luther, Northeastern Univ. (United States)
Livia Mendes, Northeastern Univ. (United States)
Jiayi Pan, Northeastern Univ. (United States)
Daniel Costa, Northeastern Univ. (United States)
Can Sarisozen, Northeastern Univ. (United States)
Vladimir Torchilin, Northeastern Univ. (United States)

Published in SPIE Proceedings Vol. 10503:
Quantitative Phase Imaging IV
Gabriel Popescu; YongKeun Park, Editor(s)

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