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Fluorescence In Situ Hybridization with Concomitant Immunofluorescence in Human Pancreas

Jody Ye1, Kathleen M Gillespie2

  • 1Diabetes and Metabolism Unit, School of Clinical Sciences, Southmead Hospital, University of Bristol, Bristol, UK.

Methods in Molecular Biology (Clifton, N.J.)
|December 15, 2015
PubMed

Insights

This study presents a new method for simultaneously identifying non-host cells and their phenotypes in pancreatic tissue. This technique is crucial for advancing research in transplantation and pregnancy-related microchimerism studies.

Area of Science:

  • Cell Biology
  • Genetics
  • Immunology

Background:

  • Identifying non-host cells in human pancreas is vital for transplantation and microchimerism research.
  • Current methods face challenges due to epitope loss during tissue processing for fluorescence in situ hybridization (FISH).
  • Immunofluorescence antigen retrieval can interfere with FISH analysis.

Purpose of the Study:

  • To develop a robust method for simultaneous analysis of X/Y chromosome constitution and cell phenotype.
  • To overcome limitations of existing techniques in pancreatic tissue analysis.

Main Methods:

  • Simultaneous fluorescence in situ hybridization (FISH) and immunofluorescence.
  • Tissue processing optimized to preserve epitopes for both techniques.
  • Analysis of X/Y chromosome constitution and cell phenotype on the same tissue section.

Main Results:

  • A validated method for concurrent detection of chromosome content and cell phenotype.
  • Successful preservation of epitopes for immunofluorescence after FISH-compatible processing.
  • Demonstration of simultaneous analysis on pancreatic tissue sections.

Conclusions:

  • The described method enables robust, simultaneous analysis of chromosome constitution and cell phenotype in pancreatic tissue.
  • This technique facilitates studies of transplantation and microchimerism.
  • It overcomes technical challenges associated with combining FISH and immunofluorescence.

Keywords:
Fluorescence in situ hybridizationMicrochimerismTransplantation

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