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Combined Lectin- and Immuno-histochemistry (CLIH) for Fluorescence Microscopy

Daša Zupančič1, Mateja Erdani Kreft2, Rok Romih2

  • 1Institute of Cell Biology, Faculty of Medicine, University of Ljubljana, Ljubljana, Slovenia. dasa.zupancic@mf.uni-lj.si.

Methods in Molecular Biology (Clifton, N.J.)
|September 24, 2022
PubMed

Insights

Combined lectin- and immuno-histochemistry (CLIH) enhances glycoprotein analysis in tissues. This method integrates immunohistochemistry and lectin histochemistry for improved glycobiology research and cancer classification.

Area of Science:

  • Glycobiology
  • Cell Biology
  • Histochemistry

Background:

  • Glycoprotein function relies on both polypeptide chains and sugar residues.
  • Immunohistochemistry (IHC) and lectin histochemistry (LHC) are standard but separate methods for glycoprotein detection.
  • Alterations in glycoprotein glycosylation are linked to cancer transformation.

Purpose of the Study:

  • To introduce combined lectin- and immuno-histochemistry (CLIH) for analyzing glycoprotein expression and distribution.
  • To demonstrate the utility of CLIH in glycobiology and cell biology research.
  • To explore CLIH's potential for improved cancer classification.

Main Methods:

  • Detailed description of five CLIH protocols.
  • Application of CLIH on paraffin sections and semithin cryosections.
  • Utilizing fluorescence microscopy for CLIH analysis.
  • Inclusion of appropriate negative controls for validation.

Main Results:

  • CLIH effectively combines the strengths of IHC and LHC.
  • The method allows for a more comprehensive understanding of glycoprotein variants.
  • CLIH protocols are adaptable for different tissue types and microscopy techniques.

Conclusions:

  • CLIH offers a powerful approach for detailed glycoprotein analysis.
  • This technique can advance research in glycobiology and cell biology.
  • CLIH holds promise for enhancing cancer diagnostics and classification.

Keywords:
AntibodiesCombined lectin- and immuno-histochemistry (CLIH)GlycoproteinsImmunohistochemistryLectin histochemistryLectinsParaffin sectionsSemithin cryosectionsSugar residues

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