Macromolecular diffusion in the extracellular matrix measured by fluorescence correlation spectroscopy

Diffusion of therapeutic macromolecules through the extracellular matrix of tumor tissue is a crucial step in drug delivery. We use fluorescence correlation spectroscopy (FCS) to measure diffusion of IgG and dextrans ( and ) in solution, 5 gelatin hydrogel, and multicellular spheroids. Gel and spher...

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Bibliographic Details
Published in:Journal of Biomedical Optics 2008-09, Vol.13 (5), p.054040-054049
Main Authors: Reitan, Nina Kristine, Juthajan, Aphirak, Lindmo, Tore, de Lange Davies, Catharina
Format: Article
Language:English
Subjects:
Algorithms
biology
Cell Line
confocal optics
correlation
Correlation analysis
Diffusion
Drug delivery systems
Extracellular Matrix - chemistry
Fluorescence
fluorescence spectroscopy
Humans
Macromolecular Substances - chemistry
Macromolecules
Microscopy, Fluorescence - methods
Spectrometry, Fluorescence - methods
Spectroscopy
Spheroids
Spheroids, Cellular - chemistry
tissues
Tumors
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Summary:Diffusion of therapeutic macromolecules through the extracellular matrix of tumor tissue is a crucial step in drug delivery. We use fluorescence correlation spectroscopy (FCS) to measure diffusion of IgG and dextrans ( and ) in solution, 5 gelatin hydrogel, and multicellular spheroids. Gel and spheroids are used as model systems for the extracellular matrix. The diffusion depends on the complexity of the environment, as well as on the size and structural shape of the diffusing molecules. The results based on one-photon FCS are in good agreement with diffusion coefficients obtained with two-photon fluorescence recovery after photobleaching (FRAP) using the same microscope (Zeiss LSM510 META/Confocor2). However, FCS reveals anomalous or multicomponent diffusion in gel and spheroids, which are not resolvable with FRAP. This study demonstrates that one-photon FCS can be used to study the extracellular transport of macromolecules in tumor tissue, and that FCS provides additional information about diffusion properties compared to FRAP.
ISSN:1083-3668
1560-2281
DOI:10.1117/1.2982530