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Abstract

Medicine

Near Infrared Optical Projection Tomography for Assessments of β-cell Mass Distribution in Diabetes Research

Published: January 12th, 2013

DOI:

10.3791/50238

1Umeå Centre for Molecular Medicine, Umeå University, 2Cell Transplant Center, Diabetes Research Institute, University of Miami,, 3EMBL-CRG Systems Biology Program, Centre for Genomic Regulation, Catalan Institute of Research and Advanced Studies, 4Dept. of Computing Science, Umeå University
* These authors contributed equally

ERRATUM NOTICE

Important: There has been an erratum issued for this article. Read more …

By adapting OPT to include the capability of imaging in the near infrared (NIR) spectrum, we here illustrate the possibility to image larger bodies of pancreatic tissue, such as the rat pancreas, and to increase the number of channels (cell types) that may be studied in a single specimen. We further describe the implementation of a number of computational tools that provide: 1/ accurate positioning of a specimen's (in our case the pancreas) centre of mass (COM) at the axis of rotation (AR)2; 2/ improved algorithms for post-alignment tuning which prevents geometric distortions during the tomographic reconstruction2 and 3/ a protocol for intensity equalization to increase signal to noise ratios in OPT-based BCM determinations3. In addition, we describe a sample holder that minimizes the risk for unintentional movements of the specimen during image acquisition. Together, these protocols enable assessments of BCM distribution and other features, to be performed throughout the volume of intact pancreata or other organs (e.g. in studies of islet transplantation), with a resolution down to the level of individual islets of Langerhans.

Erratum

Erratum: Near Infrared Optical Projection Tomography for Assessments of β-cell Mass Distribution in Diabetes Research

A correction was made to Near Infrared Optical Projection Tomography for Assessments of β-cell Mass Distribution in Diabetes Research. In Protocol section 1.1, the order of the listed chemicals MeOH:H2O2:DMSO has accidentally been switched and should instead be MeOH:DMSO:H2O2.

Protocol section 1.1 was changed from:

Incubate the tissue in freshly prepared MeOH:H2O2:DMSO bleaching buffer in a 2:1:3 ratio at RT for 24 hr to quench endogenous tissue fluorescence. For larger samples, exchange to new bleaching buffer and incubate for another 24 hr.

to:

Incubate the tissue in freshly prepared MeOH:DMSO:H2O2 bleaching buffer in a 2:1:3 ratio at RT for 24 hr to quench endogenous tissue fluorescence. For larger samples, exchange to new bleaching buffer and incubate for another 24 hr.

Tags

Keywords Near Infrared Optical Projection Tomography

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