Fixation with Carnov's fluid reduces the number of chymase-positive mast cells: not all chvmase-Dositive mast cells are also posithe for iryptase

Authors

  • A. KleinJan,

    Corresponding author
    1. Departments of Otorhinolaryngology University Hospital Dijkzigt and Erasmus University, Rotterdam, The Netherlands
    2. Departments of Imrnunology, University Hospital Dijkzigt and Erasmus University, Rotterdam, The Netherlands
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  • T. Godthelp,

    1. Departments of Otorhinolaryngology University Hospital Dijkzigt and Erasmus University, Rotterdam, The Netherlands
    2. Departments of Imrnunology, University Hospital Dijkzigt and Erasmus University, Rotterdam, The Netherlands
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  • H. M. Blom,

    1. Departments of Otorhinolaryngology University Hospital Dijkzigt and Erasmus University, Rotterdam, The Netherlands
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  • W. J. Fokkens

    1. Departments of Otorhinolaryngology University Hospital Dijkzigt and Erasmus University, Rotterdam, The Netherlands
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5 Department of Otorhinolaryngology Room Ee 1655 Erasmus University Rotterdam 3015 GD Rotterdam The Netherlands

Abstract

Mast cells in the nasal mucosa can be studied by means of monoclonal antibodies (mAb) against tryptase (T+MC) and chymase (C+MC). Fixation with acetone gives more positive cells than does fixation with Carnoy's fluid. In frozen biopsy specimens of allergic nasal mucosa fixed with acetone, the number of T+MC equals that of C+MC. When fixed with Carnoy's fluid, however, the number of T+MC is larger than the number of C+MC. The decrease in both T+MC and C+MC resulting from fixation with Carnoy's fluid is time-related and depends on the type of mAb used. Carnoy fixation time gives a decrease in the number of C+MC within 1 min, whereas the number of T+MC decreases only after 10 min. Within 1 min, the number of C+MC decreases to a level where continued fixation no longer gives further decreases in the number of cells. Two populations of mast cells can be distinguished here: one sensitive and the other insensitive to Carnoy's fluid. When double-staining is used, fixation with acetone gives three populations of mast cells: one positive for tryptase (T+C-MC), another positive for tryptase and chymase (T+C+MC), and a third one positive for chymase (T-C+MC). These three populations were found in lymph node, spleen, thymus, dermis, lung parenchyma, small intestinal submucosa, and nasal mucosa.

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