Staining Protocols for Human Pancreatic Islets

Martha L. Campbell-Thompson; Tiffany Heiple; Emily Montgomery; Li Zhang; Lynda Schneider

doi:10.3791/4068

Protocolos de coloração de ilhotas pancreáticas humanas

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JoVE Journal Medicine

Staining Protocols for Human Pancreatic Islets

Protocolos de coloração de ilhotas pancreáticas humanas

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27,815 Views

07:48 min

May 23, 2012

DOI: 10.3791/4068-v

Martha L. Campbell-Thompson¹, Tiffany Heiple¹, Emily Montgomery¹, Li Zhang¹, Lynda Schneider¹

¹Department of Pathology, Immunology, and Laboratory Medicine,University of Florida

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Overview

This video demonstrates procedures for the characterization of human pancreatic islets using hematoxylin and eosin (H&E) staining and immunohistochemistry (IHC). The technique allows for the assessment of islet endocrine composition, cell replication, and inflammatory infiltrates.

Key Study Components

Area of Science

Neuroscience
Endocrinology
Histology

Background

Human pancreatic islets play a crucial role in glucose metabolism.
Characterization of islet composition is essential for understanding diabetes.
H&E and IHC are standard techniques for tissue analysis.
Maintaining the original orientation of slides is critical for accurate assessment.

Purpose of Study

To characterize human pancreatic islets.
To determine the endocrine composition of islets.
To assess cell replication and inflammatory responses.

Methods Used

Creating serial sections from paraffin-embedded pancreas samples.
Staining sections with H&E and IHC.
Scanning stained slides for analysis.
Organizing images by case for database upload.

Main Results

Identification of insulin, glucagon, and pancreatic polypeptide in islets.
Assessment of cell replication using Ki67 staining.
Evaluation of inflammatory infiltrates through H&E and CD3 staining.
Successful localization of the uncinate region using IHC.

Conclusions

The method provides a comprehensive characterization of pancreatic islets.
Maintaining original slide orientation enhances analysis accuracy.
Results contribute to the understanding of pancreatic function in health and disease.

Frequently Asked Questions

What is the significance of characterizing pancreatic islets?

Characterizing pancreatic islets is essential for understanding their role in glucose metabolism and diabetes.

How does immunohistochemistry contribute to this study?

Immunohistochemistry allows for the specific identification of hormones and cell types within the islets.

What are the advantages of using H&E staining?

H&E staining provides a clear overview of tissue morphology and cellular structure.

Why is maintaining slide orientation important?

Maintaining slide orientation ensures accurate localization and assessment of islet structures.

What can be done with the scanned images?

Scanned images can be uploaded to online pathology databases for further research and collaboration.

Este vídeo demonstra os procedimentos para caracterização de ilhotas pancreáticas humanas com hematoxilina e eosina (H & E) e imuno-histoquímica (IHQ). Seções pancreáticas de cabeça, corpo e regiões da cauda estão manchadas por tanto H & E e IHC para determinar a composição endócrino ilhéu (insulina, glucagon e polipeptídeo pancreático), a replicação celular (Ki67), e infiltrado inflamatório (H & E, CD3). A região uncinado está localizado usando IHC para polipeptídeo pancreático.

O objetivo geral deste procedimento é caracterizar ilhós pancreáticos humanos usando coloração de hematina e eosina e imuno-histoquímica. Isso é feito criando seções seriais a partir de uma amostra de pâncreas embutida em um bloco de parafina. As seções são então colocadas em slides em sua orientação original.

Uma parte das lâminas é corada com hematina e eoína, enquanto as lâminas restantes são coradas por imuno-histoquímica, a etapa final é escanear as lâminas coradas e organizá-las por caso. Em última análise, essas imagens podem ser carregadas em um banco de dados de patologia online, onde podem ser visualizadas por outros pesquisadores da área. A principal vantagem dessa técnica sobre os métodos existentes é que as lâminas são coradas em série, mantendo sua orientação original.

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