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The use of fluorescence correlation spectroscopy (FCS) as an alternative biomarker detection technique: a preliminary study.

Shahzad A, Knapp M, Lang I, Köhler G - J. Cell. Mol. Med. (2011)

Bottom Line: Because diffusion speed is correlated with shape and molecular mass of the fluorescent molecule, this property makes it possible to study the complex formation between a small fluorescently labelled and a large unlabelled molecule.In this preliminary study, we utilize this FCS property for detection of serum biomarker.Further studies on various pathological serum samples are warranted to explore further aspects of this technique.

View Article: PubMed Central - PubMed

Affiliation: Max F. Perutz Laboratories, Department of Structural Biology and Biomolecular Chemistry, University of Vienna, Vienna, Austria. aamir.shahzad@univie.ac.at

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Typical ACF of IL-8 Antibody-DyLight488 in diluted serum.
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fig02: Typical ACF of IL-8 Antibody-DyLight488 in diluted serum.

Mentions: The number and diffusion coefficient of fluorescent particles which diffuse through the focus volume are extracted by application of the ACF. Increase in diffusion time was observed for antigen–antibody complex in both pure and diluted serum samples as compared to diffusion time of only labelled antibody (Figs 2–4). However, increase in diffusion time was more significant in case of diluted serum samples with 50% PBS (v/v = 1:1) as compared to pure serum samples. Significant increase in diffusion time was also observed after addition of the second antibody as compared to only antigen–single antibody complex in diluted serum samples (Figs 3 and 4). We found no difference in diffusion time irrespective of incubation times. Overnight incubation yielded same results as compared to 30 min. incubation time. We found same results with 10 and 20 sec. recording times. We propose that dilution of serum sample with PBS decreases viscosity of serum and is more suitable for this FCS technique as compared to pure serum.


The use of fluorescence correlation spectroscopy (FCS) as an alternative biomarker detection technique: a preliminary study.

Shahzad A, Knapp M, Lang I, Köhler G - J. Cell. Mol. Med. (2011)

Typical ACF of IL-8 Antibody-DyLight488 in diluted serum.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4373439&req=5

fig02: Typical ACF of IL-8 Antibody-DyLight488 in diluted serum.
Mentions: The number and diffusion coefficient of fluorescent particles which diffuse through the focus volume are extracted by application of the ACF. Increase in diffusion time was observed for antigen–antibody complex in both pure and diluted serum samples as compared to diffusion time of only labelled antibody (Figs 2–4). However, increase in diffusion time was more significant in case of diluted serum samples with 50% PBS (v/v = 1:1) as compared to pure serum samples. Significant increase in diffusion time was also observed after addition of the second antibody as compared to only antigen–single antibody complex in diluted serum samples (Figs 3 and 4). We found no difference in diffusion time irrespective of incubation times. Overnight incubation yielded same results as compared to 30 min. incubation time. We found same results with 10 and 20 sec. recording times. We propose that dilution of serum sample with PBS decreases viscosity of serum and is more suitable for this FCS technique as compared to pure serum.

Bottom Line: Because diffusion speed is correlated with shape and molecular mass of the fluorescent molecule, this property makes it possible to study the complex formation between a small fluorescently labelled and a large unlabelled molecule.In this preliminary study, we utilize this FCS property for detection of serum biomarker.Further studies on various pathological serum samples are warranted to explore further aspects of this technique.

View Article: PubMed Central - PubMed

Affiliation: Max F. Perutz Laboratories, Department of Structural Biology and Biomolecular Chemistry, University of Vienna, Vienna, Austria. aamir.shahzad@univie.ac.at

Show MeSH