Measurement of apoptosis of intact human islets by confocal optical sectioning and stereologic analysis of YO-PRO-1-stained islets Journal Article

  

Authors:	Boffa, D. J.; Waka, J.; Thomas, D.; Suh, S.; Curran, K.; Sharma, V. K.; Besada, M.; Muthukumar, T.; Yang, H.; Suthanthiran, M.; Manova, K.
Article Title:	Measurement of apoptosis of intact human islets by confocal optical sectioning and stereologic analysis of YO-PRO-1-stained islets
Abstract:	Apoptosis is an established pathway for islet cell demise. Current protocols for assessment of islet cell apoptosis are time-consuming (as with terminal deoxynucleotide transferase-mediated dUTP nick-end labeling reaction) and involve disruption of the islet architecture (as with flow cytometry) or destruction of cell integrity (as with enzyme-linked immunosorbent assay). The membranes of apoptotic cells, but not those of live cells, are permeant to the DNA-intercalant dye YO-PRO-1. We report a novel methodology for the rapid quantification of apoptosis of human islets: confocal laser optical sectioning and stereologic analysis of intact human islets stained with YO-PRO-1 and Hoechst 33342. The advantages include (1) rapid quantification of apoptosis without disrupting islet architecture and (2) identification of significant heterogeneity in the extent of apoptosis among islets from the same isolate. Confocal laser scanning microscopy microscopic imaging of YO-PRO-1-stained islets may advance investigation of islet cell apoptosis and help develop islet parameters predictive of posttransplant function. Copyright © 2005 by Lippincott Williams & Wilkins.
Keywords:	controlled study; human cell; apoptosis; microscopy, confocal; fluorescent dye; fluorescent dyes; cell heterogeneity; confocal laser microscopy; staining and labeling; stereology; cytoarchitecture; islets of langerhans; pancreas islet cell; islets; yo-pro-1; stereometry; benzoxazoles; quinolinium compounds
Journal Title:	Transplantation
Volume:	79
Issue:	7
ISSN:	0041-1337
Publisher:	Lippincott Williams & Wilkins
Date Published:	2005-04-15
Start Page:	842
End Page:	845
Language:	English
DOI:	10.1097/01.tp.0000155175.24802.73
PUBMED:	15818328
PROVIDER:	scopus
DOI/URL:	http://www.scopus.com/inward/record.url?eid=2-s2.0-20244385008&partnerID=40&md5=56c58c0c57863980ef755f37bd0f030a
Notes:	--- - "Cited By (since 1996): 12" - "Export Date: 24 October 2012" - "CODEN: TRPLA" - "Source: Scopus"

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