Introduction: To date a reliable method to correlate degranulation-related depletion of histochemically evidenced components of mast cell cytoplasm to intestine contraction is lacking. Degranulation of intestinal mast cells in rainbow trout was studied ex vivo by means of image analysis technique and correlated to the maximal intestinal contraction elicited by degranulation itself. Materials and Methods: Two strips from the same intestinal segment from ten trout were sampled. One was immediately mounted in an organ bath, to test the effect of incremental doses of compound 48/80, then processed, as the remnant untreated strip, for light microscopy (Giemsa stained). Color pictures from each section were converted in their gray levels RGB stacks equivalent and in eight bits gray levels. Five granular cytoplasm areas of mast cells for each section were analyzed for mean gray values. Results and Conclusion: Differential mean gray values (after compound 48/80 - before compound 48/80) of Green (R2 0.84, p < 0.01) and Blue (R2 0.83, p < 0.01) channels and eight bits grayscale (R2 0.76, p < 0.05) correlated significantly with the respective value of maximal intestinal contraction, "controlling for" the effect of the mean gray value before compound 48/80 addition. Basing upon the results, a possible acidic/anionic pro-contractile agonist can be inferred. The proposed ex vivo model and RGB stacks gray-level analysis represent a promising tool to study the role of piscine mast cell in intestine contractile modulation.[...]

RGB stacks gray-level analysis to study mast cell degranulation to intestinal contraction correlation in trout

MANERA, Maurizio
2013

Abstract

Introduction: To date a reliable method to correlate degranulation-related depletion of histochemically evidenced components of mast cell cytoplasm to intestine contraction is lacking. Degranulation of intestinal mast cells in rainbow trout was studied ex vivo by means of image analysis technique and correlated to the maximal intestinal contraction elicited by degranulation itself. Materials and Methods: Two strips from the same intestinal segment from ten trout were sampled. One was immediately mounted in an organ bath, to test the effect of incremental doses of compound 48/80, then processed, as the remnant untreated strip, for light microscopy (Giemsa stained). Color pictures from each section were converted in their gray levels RGB stacks equivalent and in eight bits gray levels. Five granular cytoplasm areas of mast cells for each section were analyzed for mean gray values. Results and Conclusion: Differential mean gray values (after compound 48/80 - before compound 48/80) of Green (R2 0.84, p < 0.01) and Blue (R2 0.83, p < 0.01) channels and eight bits grayscale (R2 0.76, p < 0.05) correlated significantly with the respective value of maximal intestinal contraction, "controlling for" the effect of the mean gray value before compound 48/80 addition. Basing upon the results, a possible acidic/anionic pro-contractile agonist can be inferred. The proposed ex vivo model and RGB stacks gray-level analysis represent a promising tool to study the role of piscine mast cell in intestine contractile modulation.[...]
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11575/4085
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