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A microfluidic imaging chamber for the direct observation of chemotactic transmigration

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Abstract

To study the roles of nonmuscle myosin II (NM-II) during invasive cell migration, microfluidic migration chambers have been designed and fabricated using photo- and soft-lithography microfabrication techniques. The chamber consists of two channels separated by a vertical barrier with multiple bays of pores with widths varying from 6 µm to 16 µm, and lengths varying from 25 µm to 50 µm. The cells are plated in the channel on one side of the barrier while a chemoattractant is flowed through the channel on the other side of the barrier. In these chambers, cells can be observed with transmitted light or fluorescence optics while they chemotax through various sized pores that impose differential mechanical resistance to transmigration. As an initial test of this device, we compared breast-cancer cell chemotactic transmigration through different pore sizes with and without inhibition of NM-II. Two distinct rates were observed as cells attempted to pull their nucleus through the smaller pores, and the faster nuclear transit mode was critically dependent on NM-II motor activity. The ability to monitor cells as they chemotax through pores of different dimensions within a single experimental system provides novel information on how pore size affects cell morphology and migration rate, providing a dramatic improvement of imaging potential relative to other in vitro transmigration systems such as Boyden chambers.

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Acknowledgment

The authors thank Wan-Hsiang Liang for help with SEM. The research is supported by a NIH grant (EB006203) to H. Baskaran and a NIH grant (GM077224) to T. Egelhoff.

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Authors and Affiliations

  1. Department of Chemical Engineering, Case Western Reserve University, 10900 Euclid Ave., Cleveland, OH, 44106, USA

    Harihara Baskaran

  2. Department of Physiology and Biophysics, Case Western Reserve University School of Medicine, 10900 Euclid Ave., Cleveland, OH, 44106, USA

    Mark T. Breckenridge

  3. Department of Cell Biology, Cleveland Clinic Foundation, 9500 Euclid Avenue, Cleveland, OH, 44195, USA

    Mark T. Breckenridge & Thomas T. Egelhoff

Authors
  1. Mark T. Breckenridge
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  2. Thomas T. Egelhoff
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  3. Harihara Baskaran
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Corresponding author

Correspondence to Harihara Baskaran.

Additional information

Thomas T. Egelhoff and Harihara Baskaran contributed equally, and are co-senior authors on this publication.

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Online Resource 1

Three ‘fast’ cells can be seen migrating towards and through a 50 × 6 μm pore (0–4 s). A ‘slow’ cell can be seen extending its leading edge towards the pore starting at ∼3 s, and migrating through the pore between seconds 5–8. Movie displayed at 20 frames sec-1, made from 17 h timelapse experiment, images taken every 5 min (MPG 3468 kb)

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Breckenridge, M.T., Egelhoff, T.T. & Baskaran, H. A microfluidic imaging chamber for the direct observation of chemotactic transmigration. Biomed Microdevices 12, 543–553 (2010). https://doi.org/10.1007/s10544-010-9411-8

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  • DOI: https://doi.org/10.1007/s10544-010-9411-8

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