Quantifying anisotropic solute transport in protein crystals using 3-D laser scanning confocal microscopy visualization

A. Cvetkovic, A. J.J. Straathof, D. N. Hanlon, S. Van Der Zwaag, R. Krishna, L. A.M. Van Der Wielen

Research output: Contribution to journalArticlepeer-review

Abstract

The diffusion of a solute, fluorescein into lysozyme protein crystals has been studied by confocal laser scanning microscopy (CLSM). Confocal laser scanning microscopy makes it possible to non-invasively obtain high-resolution three-dimensional (3-D) images of spatial distribution of fluorescein in lysozyme crystals at various time steps. Confocal laser scanning microscopy gives the fluorescence intensity profiles across horizontal planes at several depths of the crystal representing the concentration profiles during diffusion into the crystal. These intensity profiles were fitted with an anisotropic model to determine the diffusivity tensor. Effective diffusion coefficients obtained range from 6.2 × 10-15 to 120 × 10 -15 m2/s depending on the lysozyme crystal morphology. The diffusion process is found to be anisotropic, and the level of anisotropy depends on the crystal morphology. The packing of the protein molecules in the crystal seems to be the major factor that determines the anisotropy.

Original languageEnglish
Pages (from-to)389-398
Number of pages10
JournalBiotechnology and Bioengineering
Volume86
Issue number4
DOIs
Publication statusPublished - 20 May 2004
Externally publishedYes

Keywords

  • Anisotropy
  • CLSM
  • Diffusion
  • Protein crystals

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