Abstract: |
Live-cell analysis of individual cells is a challenging endeavor due to the small volume of a single cell, cell motility and the complexity of cellular systems.
Novel methodologies and means for analyzing dynamic states of individual cells/cell groups which enable monitoring alteration in cell behavior and function over time in a complex tissue environment, and assessing functional changes at single-element level are presented.
The platforms are based on various types of arrays made of high-quality transparent miniature femtoliter-, picoliter- and nanoliter-volume vessels, for investigating living cells at molecular, cellular and multicellular levels, respectively.
Each pico/nano-liter chamber is designed to hold a single cell or cell-aggregate without tethering. The technology is extremely flexible, enabling use of a spectrum of biomaterials and control over dimension, shape, configuration and distribution of the microstructures, and thus, facilitating adaptation of the arrays to various cell types and use of both label-free and fluorescence detection methods to determine live-cell status in real time.
Practical applications of the platforms include: generation and drug screening of 3D multicellular cancer spheroids, multiplexed analysis of cancer stem cells and tumor heterogeneity, measurement of biochemical processes in live cells, followed by cell content analysis of the same cells, cryopreservation of individual cells, single cell analysis of cell transfection and protein translocation.
The methodology, together with quantitative image analysis at single-element resolution, may be applicable in personalized cancer treatment, including multiplex ex-vivo analysis of heterogeneous patient-derived specimens, and for detection and evaluation of drug sensitivity of specific cell phenotypes.
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