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Journal of Biomedical Optics • Open Access

Spatial cluster analysis of nanoscopically mapped serotonin receptors for classification of fixed brain tissue
Author(s): Michael Sams; Rene Silye; Janett Gohring; Leila A. Muresan; Kurt Schilcher; Jaroslaw Jacak

Paper Abstract

We present a cluster spatial analysis method using nanoscopic dSTORM images to determine changes in protein cluster distributions within brain tissue. Such methods are suitable to investigate human brain tissue and will help to achieve a deeper understanding of brain disease along with aiding drug development. Human brain tissue samples are usually treated postmortem via standard fixation protocols, which are established in clinical laboratories. Therefore, our localization microscopy-based method was adapted to characterize protein density and protein cluster localization in samples fixed using different protocols followed by common fluorescent immunohistochemistry techniques. The localization microscopy allows nanoscopic mapping of serotonin 5-HT1A receptor groups within a two-dimensional image of a brain tissue slice. These nanoscopically mapped proteins can be confined to clusters by applying the proposed statistical spatial analysis. Selected features of such clusters were subsequently used to characterize and classify the tissue. Samples were obtained from different types of patients, fixed with different preparation methods, and finally stored in a human tissue bank. To verify the proposed method, samples of a cryopreserved healthy brain have been compared with epitope-retrieved and paraffin-fixed tissues. Furthermore, samples of healthy brain tissues were compared with data obtained from patients suffering from mental illnesses (e.g., major depressive disorder). Our work demonstrates the applicability of localization microscopy and image analysis methods for comparison and classification of human brain tissues at a nanoscopic level. Furthermore, the presented workflow marks a unique technological advance in the characterization of protein distributions in brain tissue sections.

Paper Details

Date Published: 2 December 2013
PDF: 12 pages
J. Biomed. Opt. 19(1) 011021 doi: 10.1117/1.JBO.19.1.011021
Published in: Journal of Biomedical Optics Volume 19, Issue 1
Show Author Affiliations
Michael Sams, Upper Austria Univ. of Applied Sciences (Austria)
Rene Silye, Nerve Clinic Linz Wagner Jauregg (Austria)
Janett Gohring, Medizinische Univ. Wien (Austria)
Leila A. Muresan, Ecole Normale Supérieure (France)
Kurt Schilcher, Upper Austria Univ. of Applied Sciences (Austria)
Jaroslaw Jacak, Upper Austria Univ. of Applied Sciences (Austria)

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