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Method for the determination of mean densitometric profiles of chromosomes

Application to human chromosomes stained by quinacrine mustard, ethidium bromide or by the Feulgen reaction

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Abstract

When comparing the densitometric profiles of corresponding chromosomes registered from different metaphases or homologous pairs, one is always faced with the variability of their length and overall height. This makes difficult the quantitative comparison of a given chromosome treated by various staining procedures. — A simple and rapid method has been developed for normalizing the densitometric profiles and averaging them in order to obtain a “mean density pattern” of each chromosome. The analysis involves: photographic images, digitalization of the densitometric profiles and processing of the data by a mini-computer. — The method, based on a linear relationship between the area of the densitometric profiles and their length, has been applied to five human chromosomes (1, 2, 6, 12 and 16) stained by ethidium bromide, quinacrine mustard (with or without acidic hydrolysis), pararosaniline and bisaminophenyl-oxadiazole (Feulgen reaction).

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References

  • Andersson, G. K. A., Kjellstrand, P. T. T.: Exposure and removal of stainable groups during Feulgen acid hydrolysis of fixed chromatin at different temperatures. Histochemie 27, 165–172 (1971)

    Google Scholar 

  • Bailly, S.: Etude cytophotométrique de la distribution des acides nucléiques le long des chromosomes métaphasiques de l'Amphibien Urodèle Pleurodeles waltlii Michah. Ann. Génét. 12, 184–190 (1969)

    Google Scholar 

  • Bedi, K. S., Goldstein, D. J.: Cytophotometric factors causing apparent differences between Feulgen DNA contents of different leucocytes types. Nature (Lond.) 251, 439–440 (1974)

    Google Scholar 

  • Böhm, N., Sprenger, E.: Fluorescence cytophotometry: a valuable method for the quantitative determination of nuclear FeuIgen-DNA. Histochemie 16, 100–118 (1968)

    Google Scholar 

  • Caspersson, T., Farber, S., Foley, G. E., Kudynowski, J., Modest, E. J., Simonsson, E., Wagh, U., Zech, L.: Chemical differentiation along metaphase chromosomes. Exp. Cell Res. 49, 219–222 (1968)

    Google Scholar 

  • Caspersson, T., Lomakka, G., Møller, A.: Computerized chromosomes identification by aid of the quinacrine mustard fluorescence technique. Hereditas (Lund) 67, 103–110 (1971a)

    Google Scholar 

  • Caspersson, T., Lomakka, G., Zech, L.: The 24 fluorescence patterns of the human metaphase chromosomes-distinguishing characters and variability. Hereditas (Lund) 67, 89–102 (1971b)

    Google Scholar 

  • Caspersson, T., Zech, L., Johansson, C., Modest, E. J.: Identification of human chromosomes by DNA reacting fluorescing agents. Chromosoma (Berl.) 30, 215–227 (1970)

    Google Scholar 

  • Castelman, K. R., Wall, R. J.: Automatic systems for chromosome identification. In: Chromosome identification (T. Caspersson and L. Zech, eds.), p. 77–84. New York, London: Academic Press 1973

    Google Scholar 

  • Deitch, A. D., Wagner, D., Richart, R. M.: Conditions influencing the intensity of the Feulgen reaction. J. Histochem. Cytochem. 16, 371–379 (1968)

    Google Scholar 

  • Distèche, C., Hagemeijer, A. M., Frederic, J., Progneaux, D.: An abnormal large human chromosome identified as an end-to-end fusion of two X's by combined results of the new banding techniques and microdensitometry. Clin. Genet. 3, 388–395 (1972)

    Google Scholar 

  • Fitzgerald, P. H.: Differential contraction of large and small chromosomes in cultured leucocytes of man. Cytogenetics 4, 65–73 (1965)

    Google Scholar 

  • Gaillard, J. L. J., Vanduijn, P., Schaberg, A.: Photometric determination of DNA in human chromosomes. Exp. Cell Res. 53, 417–431 (1968)

    Google Scholar 

  • Garcia, A. M.: Studies on DNA in leucocytes and related cells of mammals. VI. The Feulgen-DNA content of Rabbit leukocytes after hypotonic treatment. J. Histochem. Cytochem. 17, 47–55 (1969a)

    Google Scholar 

  • Garcia, A. M.: Cytophotometric studies on haploid and diploid cells with different degrees of chromatin coiling. Ann. N.Y. Acad. Sci. 157, 237–249 (1969b)

    Google Scholar 

  • Gledhill, B. L., Gledhill, M. P., Rigler, R., Jr., Ringertz, N. R.: Changes in deoxyribonucleoprotein during spermiogenesis in the bull. Exp. Cell Res. 41, 652–665 (1966)

    Google Scholar 

  • Goldstein, D. J.: Aspects of scanning microdensitometry. II. Spot size, focus and resolution. J. Micr. 93, 15–42 (1971)

    Google Scholar 

  • Granlund, G. H.: The use of distribution functions to describe integrated profiles of human chromosomes. In: Chromosome identification (T. Caspersson and L. Zech, eds.), p. 85–87. New York, London: Academic Press 1973

    Google Scholar 

  • Graumann, W.: Zur Standardisierung des Schiff'sehen Reagens. Z. wiss. Mikr. 61, 225–226 (1953)

    Google Scholar 

  • Heneen, W. K., Caspersson, T.: Identification of the chromosomes of rye by distribution patterns of DNA. Hereditas (Lund) 74, 259–272 (1973)

    Google Scholar 

  • Hilditch, C. J., Rutovitz, D.: Normalization of chromosome measurements. Comput. Biol. Med. 2, 167–179 (1972)

    Google Scholar 

  • Ledley, R. S., Lubs, H. A., Ruddle, F. H.: Introduction to chromosome analysis. Comput. Biol. Med. 2, 107–128 (1972)

    Google Scholar 

  • Lubs, H. A., Ledley, R. S.: Automated analysis of differentially stained human chromosomes. In: Chromosome identification (T. Caspersson and L. Zech, eeds.), p. 61–76. New York, London: Academic Press 1973

    Google Scholar 

  • Mainguy, P. N. R., Heddle, J. A., Brunette, D. M.: A reliable method of quantifying G-band position in chromosomes. Chromosoma (Berl.) 50, 309–312 (1975)

    Google Scholar 

  • Marimuthu, K. M., Selles, W., Neurath, P. W.: Computer analysis of Giemsa banding patterns and automatic classification of human chromosomes. Amer. J. hum. Genet. 26, 369–378 (1974)

    Google Scholar 

  • Mayall, B. H., Mendelsohn, M. L.: Deoxyribonucleic acid cytophotometry of stained human leukocytes. II. The mechanical, scanner of cydac, the theory of scanning photometry and the magnitude of residual errors Histochem. Cytochem. 18, 383–407 (1970)

    Google Scholar 

  • Mendelsohn, M. L., Hungerford, D. A., Mayall, B. H., Perry, B. H., Conway, T., Prewitt, J. M. S.: Computer-oriented analysis of human chromosomes. II. Integrated optical density as a single parameter for karyotype analysis Ann. N.Y. Acad. Sci. 157, 376–392 (1969)

    Google Scholar 

  • Mendelsohn, M. L., Mayall, B. H.: Chromosome identification by image analysis and quantitative cytochemistry. In: Human chromosome methodology (J. J. Yunis, ed.), 2nd édit. p. 311–346. New York, London: Academic Press 1974

    Google Scholar 

  • Mendelsohn, M. L., Mayall, B. H., Bogart, E., Moore II, D. M., Perry, B. H.: DNA content and DNA-base centromeric index of the 24 human chromosomes. Science 179, 1126–1129 (1973)

    Google Scholar 

  • Moore II, D. M.: Do homologous chromosomes differ? Two statistical tests. Cytogenet. Cell Genet. 12, 305–314 (1973)

    Google Scholar 

  • Neurath, P. W., Kess, B., Low, D. A.: Individualized human karyotyping through quantitative analysis. Comput. Biol. Med. 2, 181–193 (1972a)

    Google Scholar 

  • Neurath, P. W., Lin, P. S., Low, D. A.: Quantitative karyotyping: a model twin study. Cytogenetics 11, 457–474 (1972b)

    Google Scholar 

  • Paris Conference (1971): Standardization in human Cytogenetics. Cytogenetics 11, 313–362 (1972)

    Google Scholar 

  • Puck, T. T., Cieciura, S. J., Robinson, A.: Long term cultivation of euploid cells from human and animal subjects. J. exp. Med. 108, 945–946 (1958)

    Google Scholar 

  • Rasch, R. W., Rasch, E. M.: Kinetics of hydrolysis during the Feulgen reaction for deoxyribonucleic acid: a reevaluation. J. Histochem. Cytochem. 21, 1053–1065 (1973)

    Google Scholar 

  • Ringertz, N. R.: Cytochemical properties of nuclear proteins and deoxyribonucleoprotein complexes in relation to nuclear function. In: Handbook of molecular cytology (A. Lima-de-Faria, ed.), p. 656–684. New York: American Elsevier Publ. Co. Inc. 1969

    Google Scholar 

  • Ruch, F.: Determination of DNA content by microfluorometry. In: Introduction to quantitative cytochemistry (G. L. Wied, ed.), p. 281–294. New York, London: Academic Press 1966

    Google Scholar 

  • Rudkin, G. T., Hungerford, D. A., Nowell, P. C.: DNA content of chromosome Phi and chromosome 21 in human chronic granulocytic leukaemia. Science 144, 1229–1232 (1964)

    Google Scholar 

  • Rudkin, G. T.: Photometric measurements of individual metaphase chromosomes. In Vitro 1, 12–20 (1965)

    Google Scholar 

  • Sasaki, M.: Observations on the modification in size and shape of chromosomes due to technical procedure. Chromosoma (Berl.) 11, 514–522 (1961)

    Google Scholar 

  • Ploeg, M. van der, Duijn, P. van, Ploem, J. S.: High-resolution scanning-densitometry of photographic negatives of human metaphases chromosomes. I. Instrumentation. Histochemistry 42, 9–30 (1974a)

    Google Scholar 

  • Ploeg, M. van der, Duijn, P. van, Ploem, J. S.: High-resolution scanning-densitometry of photographic negatives of human metaphases chromosomes. II. Feulgen-DNA measurements. Histochemistry 42, 31–46 (1974b)

    Google Scholar 

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Authors and Affiliations

  1. Laboratoire de Cytogénétique, Université de Liège, Rue des Bonnes Villes, 56, B-4020, Liège, Belgique

    C. Distèche

  2. Laboratoire de Chimie-Physique, Université de Liège au Sart-Tilman, B-4000, Liège, Belgique

    J. Bontemps

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  1. C. Distèche
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Distèche, C., Bontemps, J. Method for the determination of mean densitometric profiles of chromosomes. Chromosoma 54, 39–59 (1976). https://doi.org/10.1007/BF00331832

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  • DOI: https://doi.org/10.1007/BF00331832

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