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An automatic analysis method for in situ hybridization using high-resolution image analysis

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Summary

Specific mRNA for α1 (I) collagen has been detected on a cellular level by in situ hybridization using radioactively labelled α1 (I) antisense RNA probes. We here present an automatic, quantitative, evaluation technique for the determination of grain densities using the high-resolution image analysis system IPS KONTRON. The reliability and objectivity of this method were evaluated by comparing the values of grains per cell obtained by conventional and automatic techniques following in situ hybridization with α1 (I) collagen DNA probes in various specimens.

The correlation coefficients between conventional and automatic analysis of grain densities were 0.97 for fibroblasts embedded into a three-dimensional collagen gel, 0.94 for dermal fibroblasts in skin obtained from a patient with progressive systemic scleroderma, and 0.90 for fibroblasts in a 2-week-old scar. All correlation coefficients were significant on the P<0.0001 level. This new analysis technique therefore allows a more rapid and reliable quantitative evaluation of in situ hybridization and may also be helpful in differentiating between various cell populations characterized by different biosynthetic capacities.

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References

  1. Abmayr W, Deml E, Oesterle D, Gössner W (1983) Nuclear morphology in preneoplastic lesions of rat liver. Anal Quant Cytol 5:275–284

    Google Scholar 

  2. Abmayr W, Giaretti W, Gais P, Dörmer P (1982) Discrimination of G1, S, and G2 cells using high-resolution TV-scanning and multivariate analysis methods. Cytometry 2:316–326

    Google Scholar 

  3. Abmayr W, Stolz W, Korherr S, Wild W, Schmoeckel C (1987) Chromatin structure of melanocytic nuclei: correlation between light and electron microscopy. Applied Optics 26:3343–3348

    Google Scholar 

  4. Auffermann W, Böcking A (1985) Early detection of precancerous lesions in dysplasias of the lung by rapid DNA image cytometry. Anal Quant Cytol 7:218–226

    Google Scholar 

  5. Böcking A, Adler C-P, Common HH, Hilgarth M, Granzen B, Auffermann W (1984) Algorithm for a DNA-cytophotometric diagnosis and grading of malignancy. Anal Quant Cytol 6:1–8

    Google Scholar 

  6. Baak JPA, Oort J (1983) A manual of morphometry in diagnostic pathology. Springer, Berlin Heidelberg New York

    Google Scholar 

  7. Bartels PH, Bibbo M, Olson GB, Wied GL (1984) Cell image analysis in quantitative cytology. In: Dendy PP, Ernst DW, Senguan A (eds) Technical advances in biomedical physics. Martinus Nijhoff, The Hague, pp 59–90

    Google Scholar 

  8. Castleman KR (1979) Digital image processing. Prentice Hall, Englewood Cliffs

    Google Scholar 

  9. Chu ML, Myers J, Bernhard MP, Ding JF, Ramirez F (1982) Cloning and characterization of five overlapping cDNA specific for the human pro (I) collagen chain. Nucleic Acids Res 10:5925–5934

    Google Scholar 

  10. Cox KH, De Leon DV, Angerer LM, Angerer RC (1984) Detection of mRNAs in sea urchin embryos by in situ hybridization using asymmetric RNA probes. Develop Biol 101:485–502

    Google Scholar 

  11. Eins S (1987) Quantitative und strukturelle Bildanalyse in der Medizin. GIT, Darmstadt

    Google Scholar 

  12. Gonzalez RC, Wintz P (1978) Syntactic pattern recognition. An introduction. Addison-Wesley, Reading

    Google Scholar 

  13. Höfler H, Pütz B, Ruhri C, Wirnsberger G, Klimpfinger M, Smolle J (1987) Simultaneous localization of calcitonin mRNA and peptide in a medullary thyroid carcinoma. Virchows Arch B 54:144–151

    Google Scholar 

  14. Harms H, Aus H-M (1987) Tissue image segmentation with multicolor, multifocal algorithms. In: Devijver PA, Kittler J (eds) NATO ASI. Springer, Berlin Heidelberg New York, pp 519–527 (vol F30)

    Google Scholar 

  15. Kriete A, Schäffer R, Harms H, Aus HM (1985) Computerbased cytophotometric classification of thyroid tumors in imprints. J Cancer Res Clin Oncol 109:252–256

    Google Scholar 

  16. Kuhar MJ (1986) Quantitative receptor autoradiography: an overview. In: Boast CA, Snowhill EW, Altar CA (eds) Quantitative receptor autoradiography. Liss, New York, pp 1–12

    Google Scholar 

  17. Le Boit PE, Van Fletcher H (1987) A comparative study of Spitz nevus and nodular malignant melanoma using image analysis cytometry. J Invest Dermatol 88:753–757

    Google Scholar 

  18. Majzoub JA, Rich A, Van Boom J, Habener JF (1983) Vasopressin and oxytocin mRNA regulation in the rat assessed by hybridization with synthetic oligonucleotides. J Biol Chem 258:14061–14064

    Google Scholar 

  19. Marchevsky AM, Gil J, Jeanty H (1987) Computerized interactive morphometry in pathology: current instrumentation and methods. Hum Pathol 18:320–331

    Google Scholar 

  20. Mauch C, Hatamochi A, Scharffetter K, Krieg T (1988) Regulation of collagen synthesis in fibroblasts in a three-dimensional collagen gel. Exp Cell Res 178493–503

    Google Scholar 

  21. Nüsgens B, Merril C, Lapiere C, Bell E (1984) Collagen biosynthesis by cells in a tissue equivalent matrix in vivo. Coll Relat Res 4:351–364

    Google Scholar 

  22. Pratt WK (1978) Digital image processing. Wiley Interscience, New York

    Google Scholar 

  23. Preston K (1986) High-resolution image analysis. J Histochem Cytochem 34:67–74

    Google Scholar 

  24. Scharffetter K, Lankat-Buttgereit B, Krieg T (1988) Localization of collagen mRNA in normal and sclerodermal skin. Eur J Clin Invest 18:9–17

    Google Scholar 

  25. Scharffetter K, Stolz W, Lankat-Buttgereit C, Mauch C, Kulozik M, Krieg T (1989) In situ hybridization — a useful tool for studies on collagen gene expression in cell culture as well as in normal and altered tissue. Virchows Archiv [Cell Pathol] 56:299–306

    Google Scholar 

  26. Smolle J, Soyer HP, Kerl H (1988) Nuclear parameters of the superficial and deep portion of melanocytic lesions — a morphometric investigation. Pathol Res Pract 183:266–270

    Google Scholar 

  27. Snowhill EW, Boast CA (1986) Quantitative image analysis: application to the study of variations in regional benzodiazepine receptor binding. In: Boast CA, Snowhill EW, Altar CA (eds) Quantitative receptor autoradiography. Liss, New York, pp 13–28

    Google Scholar 

  28. Stolz W, Abmayr W, Schmoeckel C, Braun-Falco O (1987) High-resolution image analysis: a new tool for the recognition of malignant melanocytic nuclei in light microscopy. J Invest Dermatol 89:448 (abstract)

    Google Scholar 

  29. Suggs SV, Wallace RB, Hirose T, Kawashima EH, Itakura K (1981) Use of synthetic oligonucleotides as hybridization probes: isolation of cloned cDNA sequences for human beta2-microglobulin. Proc Natl Acad Sci USA 78:6613–6617

    Google Scholar 

  30. Thomas PS (1980) Hybridization of denatured RNA and small DNA fragments transferred to nitrocellulose. Proc Natl Acad Sci USA 77:5201–5205

    Google Scholar 

  31. Uhl GR, Zingg HH, Habener JF (1985) Vasopressin mRNA in situ hybridization: localization and regulation studied with oligonucleotide cDNA probes in normal and Brattleboro rat hypothalamus. Proc Natl Acad Sci USA 82:5555–5559

    Google Scholar 

  32. Wied GL, Bartels PG, Dytch HE, Bibbo M (1983) Rapid DNA evaluation in clinical diagnosis. Acta Cytol 27:33–37

    Google Scholar 

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Author notes

  1. K. Scharffetter

    Present address: Department of Dermatology, University of Düsseldorf, Moorenstraße 5, D-4000, Düsseldorf, FRG

Authors and Affiliations

  1. Department of Dermatology, University of Munich, Frauenlobstr. 9-11, 8000, München 2, FRG

    W. Stolz, K. Scharffetter & T. Krieg

  2. Fachhochschule München, Fachbereich Informatik, Infanteriestr. 14, 8000, München 2, FRG

    W. Abmayr

  3. Kontron Bildanalyse GmbH, Breslauer Str. 2, 8057, Eching bei München, FRG

    W. Köditz

Authors
  1. W. Stolz
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  2. K. Scharffetter
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  4. W. Köditz
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  5. T. Krieg
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Stolz, W., Scharffetter, K., Abmayr, W. et al. An automatic analysis method for in situ hybridization using high-resolution image analysis. Arch Dermatol Res 281, 336–341 (1989). https://doi.org/10.1007/BF00412978

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  • DOI: https://doi.org/10.1007/BF00412978

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