Generating controlled molecular gradients in 3D gels

W. J. Rosoff, R. McAllister, M. A. Esrick, G. J. Goodhill, J. S. Urbach

Research output: Contribution to journalArticlepeer-review

44 Scopus citations

Abstract

A new method for producing molecular gradients of arbitrary shape in thin three dimensional gels is described. Patterns are produced on the surface of the gel by printing with a micropump that dispenses small droplets of solution at controlled rates. The molecules in the solution rapidly diffuse into the gel and create a smooth concentration profile that is independent of depth. The pattern is relatively stable for long times, and its evolution can be accurately described by finite element modeling of the diffusion equation. As a demonstration of the method, direct measurements of protein gradients are performed by quantitative fluorescence microscopy. A complementary technique for measuring diffusion coefficients is also presented. This rapid, flexible, contactless approach to gradient generation is ideally suited for cell culture experiments to investigate the role of gradients of diffusible substances in processes such as chemotaxis, morphogenesis, and pattern formation, as well as for high-throughput screening of system responses to a wide range of chemical concentrations.

Original languageEnglish
Pages (from-to)754-759
Number of pages6
JournalBiotechnology and Bioengineering
Volume91
Issue number6
DOIs
StatePublished - Sep 20 2005

Keywords

  • Cell motility assay
  • Collagen gel
  • Diffusible factors
  • Molecular gradients

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