Staining sections of water‐miscible resins 2. Effects of staining‐reagent lipophilicity on the staining of glycol‐methacrylate‐embedded tissues

R.W. Horobin*, P.O. Gerrits, D.J. Wright

*Corresponding author for this work

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    Abstract

    Glycol methacrylate (GMA) sections of animal tissues were stained with a group of twenty-seven reagents of very varied chemical characteristics. The artefactual background staining of the resin was found to be dependent on the hydrophilic/lipophilic character of the staining reagent, as estimated from the logarithm of its octanol-water partition coefficient (log P). Intense background staining occurred with lipophilic stains, whose log P > 2. In keeping with this, use of GMA semi-permeable membranes for enzyme histochemistry failed to give staining when using a lipophilic substrate, probably because the substrate was trapped in the membrane. An analysis of other routine histochemical stains-in terms of the probable occurrence of high resin background staining and low tissue sensitivity-is made. A numerical guide is provided to help avoid artefacts resulting from hydrophobic and size effects. Note: small, hydrophilic reagents (log P <0; molecular weight <550 Da) are least likely to show either type of artefact. Conversely, reagents which are lipophilic, or/and of intermediate size (log P > 2; 550 <ionic weight <1000 Da), give strong background staining.

    Original languageEnglish
    Pages (from-to)199-205
    Number of pages7
    JournalJournal of microscopy-Oxford
    Volume166
    DOIs
    Publication statusPublished - May-1992

    Keywords

    • ARTIFACT
    • GLYCOL METHACRYLATE (GMA)
    • LIPOPHILICITY
    • STRUCTURE-STAINING CORRELATIONS
    • STRUCTURE-ACTIVITY RELATIONSHIP
    • WATER-MISCIBLE RESIN

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