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Quantifying anisotropic solute transport in protein crystals using 3‐D laser scanning confocal microscopy visualization
Author(s) -
Cvetkovic A.,
Straathof A. J. J.,
Hanlon D. N.,
van der Zwaag S.,
Krishna R.,
van der Wielen L. A. M.
Publication year - 2004
Publication title -
biotechnology and bioengineering
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.136
H-Index - 189
eISSN - 1097-0290
pISSN - 0006-3592
DOI - 10.1002/bit.20067
Subject(s) - confocal , anisotropy , confocal laser scanning microscopy , confocal microscopy , visualization , laser scanning , laser microscopy , microscopy , laser , materials science , optics , biophysics , chemistry , biology , physics , computer science , data mining
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 m 2 /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. © 2004 Wiley Periodicals, Inc.