Summary
The N-quaternized derivative of dimethyl-POPOP (termed Q4) induces a bluish-green fluorescent reaction in mast cell granules from paraffin sections and cell smears, in addition to a previously described bluish-white fluorescent reaction in chromatin DNA. The chromatin reaction was abolished by staining the samples either with Mayer's Haematoxylin before Q4 treatment or by Q4 treatment at pH 1.5. The reaction in mast cell granules was absent after substrate methylation. The staining sequence Haematoxylin-Eosin-Q4 also worked well in paraffin sections, allowing the observation of the current histological image under bright-field illumination as well as double-colour emission under fluorescence microscopy. The sequence is proposed as a new diagnostic procedure for demonstrating mast cell granules.
Similar content being viewed by others
References
Berlman, I. B. (1971) Handbook of Fluorescence Spectra of Aromatic Molecules, 2nd edn. New York: Academic Press.
Birks, J. B. (1964) The Theory and Practice of Scintillation Counting. New York: Macmillan.
Enerbäck, L. (1974) Berberine sulphate binding to mast cell polyanions: a cytofluorimetric method for the quantitation of heparin. Histochemistry 42, 301–13.
Espada, J., Valverde, P. & Stockert, J. C. (1993) Selective fluorescence of eosinophilic structures in grasshopper and mammalian testis stained with haematoxylin-eosin. Histochemistry 99, 385–90.
Espada, J., Stockert, J. C. & Gamallo, C. (1994) Fluorescencia selectiva de estructuras eosinofílicas en secciones teñidas con Hematoxilina/Eosina: mecanisos histoquímicos y aplicaciones histológicas. Patología 27, 335–41.
Fisher, E. R., Paik, S. M., Rockette, I., Jones, J., Caplan, R. & Fisher, B. and other NSABP Collaborators. (1989) Prognostic significance of eosinophils and mast cells in rectal cancer: findings from the national surgical adjuvant breast and bowel project (protocol R-01). Hum. PathoL. 20, 159–63.
Geyer, G. (1962) Der Sulfonsäurenachweis. Eine neu Methode für den histochemischen Nachweis vom Sulfonsäuren. Acta HistocheM. 14, 26–41.
Grossgebauer, K. (1979) Staining of acid mucopolysaccharides appearing on and in various cell types by DAPI. Microsc. Acta 82, 291–3.
Hadler, W. A., Cruz-Höfling, M. A. & Ziti, L. M. (1978) The benzidine technique for histochemical detection of the SO4 ion. Acta HistocheM. 61, 197–203.
Hahn von Dorsche, I. & Opitz, M. (1970) Beitrag zur Kenntnis der selektiven Fluorochromierung von Mastzellen. Acta HistocheM. 36, 74–86.
Hals, E. (1970) Some methods for fluorochromation and staining of rat mast cells with basic dyes. Scand. J. Dent. ReS. 78, 301–10.
Hartveit, F. (1981) Mast cells and metachromasia in human breast cancer: their occurrence, significance and consequence. J. PathoL. 134, 7–11.
Hebda, P. A., Collins, M. A. & Tharp, M. D. (1993) Mast cells and myofibroblasts in wound healing. Dermatol. Clinics 11, 685–96.
Love, L. D. (1979) Fluorescence microscopy of viable mast cells stained with different concentrations of acridine orange. Histochemistry 62, 221–5.
Módis, L. & Batschwarowa, M. (1969) Fluorescence histochemical investigations of connective tissue. III. Fluorescence histochemistry of the heparin content of mast cells. Acta Morph. Acad. Sci. HunG. 17, 235–40.
Murgatroyd, L. B. (1982) 2-hydroxystilbamidine isethionate: a new fluorochrome for use in general pathology. I. The selective staining of DNA, mucosubstances and elastic fibres. Histochemistry 74, 107–14.
Ott, D. G., Hayes, F. N. & Kerr, V. N. (1956) Oxazole quaternary salts. J. Am. Chem. SoC. 78, 1941–44.
Pearse, A. G. E. (1968) Histochemistry. Theoretical and Applied, 3rd edn. London: Churchill Livingstone.
Petkov, P. E. (1968) Staining and fluorescent microscopy of mast cells with pseudoisocyanine. Experientia 24, 1073.
Roche, W. R. (1985) Mast cells and tumors. The specific enhancement of tumor proliferation in vitro. Am. J. PathoL. 119, 57–64.
Roche, W. R. (1986) The nature and significance of tumour-associated mast cells. J. PathoL. 148, 175–82.
Rothe, M. J., Nowak, M. & Kardel, F. A. (1990) The mast cell in health and disease. J. Am. Acad. DermatoL. 26, 615–24.
Schwartz, L. B. (1991) Mast cells and their role in urticaria. J. Am. Acad. DermatoL. 25, 190–204.
Schwartz, L. B. (1994) Mast cells: function and contents. Curr. Op. ImmunoL. 6, 91–7.
Scott, J. E. (1973) Affinity, competition and specific interactions in the biochemistry and histochemistry of polyelectrolytes. Biochem. Soc. TranS. 1, 787–806.
Sterk, A. R. & Ishizaka, T. (1982) Binding properties of IgE receptors on normal mouse mast cells. J. ImmunoL. 128, 838–42.
Stevens, R. L. & Austen, K. F. (1989) Recent advances in the cellular and molecular biology of mast cells. Immunol. Today 10, 381–6.
Stockert, J. C. (1979) Observations on the chromatin staining by aluminium-hematoxylin. Z. NaturforscH. 34c, 1285–6.
Stockert, J. C. (1992) Fluorescence of chromatin DNA by an oxazolium scintillator. Z. NaturforscH. 47c, 481–2.
Strobel, S., Miller, H. R. P. & Ferguson, A. (1981) Human intestinal mucosal mast cells: evaluation of fixation and staining techniques. J. Clin. PathoL. 34, 851–8.
Thompson, S. M. (1966) Selected Histochemical and Histopathological Methods. Springfield, Illinois: Charles C. Thomas.
Vassar, P. S. & Culling, C. F. A. (1959) Fluorescent stains, with special reference to amyloid and connective tissue. Arch. PathoL. 68, 487–98.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Espada, J., Bertolesi, G.E., Trigoso, C.I. et al. Fluorescence of mast cell granules in paraffin sections and cell smears induced by an N-quaternary oxazole scintillator. Histochem J 27, 318–322 (1995). https://doi.org/10.1007/BF00398974
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00398974