Biomedical Microdevices

, Volume 12, Issue 3, pp 543–553 | Cite as

A microfluidic imaging chamber for the direct observation of chemotactic transmigration

  • Mark T. Breckenridge
  • Thomas T. Egelhoff
  • Harihara Baskaran
Article

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.

Keywords

Chemotaxis Transmigration Nonmuscle myosin II 

Supplementary material

10544_2010_9411_MOESM1_ESM.mpg (3.4 mb)
Online Resource 1Three ‘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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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Mark T. Breckenridge
    • 2
    • 3
  • Thomas T. Egelhoff
    • 3
  • Harihara Baskaran
    • 1
  1. 1.Department of Chemical EngineeringCase Western Reserve UniversityClevelandUSA
  2. 2.Department of Physiology and BiophysicsCase Western Reserve University School of MedicineClevelandUSA
  3. 3.Department of Cell BiologyCleveland Clinic FoundationClevelandUSA

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