NDLI: Microfluidics-integrated time-lapse imaging for analysis of cellular dynamics

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A stretching device for imaging real-time molecular dynamics of live cells adhering to elastic membranes on inverted microscopes during the entire process of the stretch

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Integrative Biology Vol - 2 Issue - 4

Integrative Biology Vol - 2 Issue - 2-3

Integrative Biology Vol -1

Microfluidics-integrated time-lapse imaging for analysis of cellular dynamics

Content Provider	Royal Society of Chemistry (RSC)
Author	Albrecht, Dirk R. Bhatia, Sangeeta N. Mendelson, Avital Shah, Jagesh V. Resnikoff, Joshua Underhill, Gregory H.
Copyright Year	2010
Abstract	An understanding of the mechanisms regulating cellular responses has recently been augmented by innovations enabling the observation of phenotypes at high spatio-temporal resolution. Technologies such as microfluidics have sought to expand the throughput of these methods, although assimilation with advanced imaging strategies has been limited. Here, we describe the pairing of high resolution time-lapse imaging with microfluidic multiplexing for the analysis of cellular dynamics, utilizing a design selected for facile fabrication and operation, and integration with microscopy instrumentation. This modular, medium-throughput platform enables the long-term imaging of living cells at high numerical aperture (via oil immersion) by using a conserved 96-well, ∼6 × 5 mm2 imaging area with a variable input/output channel design chosen for the number of cell types and microenvironments under investigation. In the validation of this system, we examined fundamental features of cell cycle progression, including mitotic kinetics and spindle orientation dynamics, through the high-resolution parallel analysis of model cell lines subjected to anti-mitotic agents. We additionally explored the self-renewal kinetics of mouse embryonic stem cells, and demonstrate the ability to dynamically assess and manipulate stem cell proliferation, detect rare cell events, and measure extended time-scale correlations. We achieved an experimental throughput of >900 cells/experiment, each observed at >40× magnification for up to 120 h. Overall, these studies illustrate the capacity to probe cellular functions and yield dynamic information in time and space through the integration of a simple, modular, microfluidics-based imaging platform.
Starting Page	278
Ending Page	287
Page Count	10
File Format	HTM / HTML PDF
ISSN	17579694
Volume Number	2
Issue Number	5-6
Journal	Integrative Biology
DOI	10.1039/b923699f
Language	English
Publisher	Royal Society of Chemistry
Access Restriction	Open
Subject Keyword	Microfluidics Time-lapse photography Microscopy Numerical aperture Oil immersion Cell cycle Laboratory mouse Stem cell
Content Type	Text
Resource Type	Article
Subject	Medicine Biochemistry Biophysics

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