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Fluorochromes for DNA Staining and Quantitation

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Histochemistry of Single Molecules

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1560))

Abstract

In these last few decades the great explosion of the molecular approaches has casted a little shadow on the DNA quantitative analysis. Nevertheless DNA cytochemistry represented a long piece of history in cell biology since the advent of the Feulgen reaction. This discovery was really the milestone of the emerging quantitative cytochemistry, and scientists from all over the world produced a very large literature on this subject. This first era of quantitation (histochemistry followed by cytochemistry) started by means of absorption measurements (histophotometry and cytophotometry). The successive introduction of fluorescence microscopy gave a great boost to quantitation, making easier and faster the determination of cell components by means of cytofluorometry. The development of flow cytometry further contributed to the importance of quantitative cytochemistry. At its beginning, the mission of flow cytometry was still DNA quantitation. For a decade the Feulgen reaction had been the reference methodology for both conventional and flow cytofluorometry; the advent of Shiff-type reagents contributed to expand the variety of possible fluorochromes excitable in the entire visible spectrum as well as in the ultraviolet region. The fluorescence scenario was progressively enriched by new probes among which are the intercalating dyes which made DNA quantitation simple and fast, thus spreading it worldwide. The final explosion of cytofluorometry was made possible by the availability of a large variety of probes directly binding DNA structure. In addition, immunofluorescence allowed to correlate the cell cycle-related DNA content to other cell markers. In the clinical application of flow cytometry, this promoted the introduction of multiparametric analyses aimed at describing the cytokinetic characteristics of a given cell subpopulation defined by a specific immunophenotype setting.

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Acknowledgments

The authors acknowledge Emanuela Cova for helpful discussion and contribution, and Carlo Pellicciari for his precious assistance in finding archival documents and pictures.

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Authors and Affiliations

  1. Institute of Molecular Genetics, CNR, Via Abbiategrasso 207, 27100, Pavia, Italy

    Giuliano Mazzini

  2. Department of Biology and Biotechnology “Lazzaro Spallanzani”, University of Pavia, Pavia, Italy

    Giuliano Mazzini

  3. Department of Medicine, Azienda Socio-Sanitaria Territoriale of Pavia, Pavia, Italy

    Marco Danova

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  1. Giuliano Mazzini
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  2. Marco Danova
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Correspondence to Giuliano Mazzini .

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Editors and Affiliations

  1. Department of Biology and Biotechnology, University of Pavia, Pavia, Italy

    Carlo Pellicciari

  2. Department of Biology and Biotechnology, University of Pavia, Pavia, Italy

    Marco Biggiogera

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Mazzini, G., Danova, M. (2017). Fluorochromes for DNA Staining and Quantitation. In: Pellicciari, C., Biggiogera, M. (eds) Histochemistry of Single Molecules. Methods in Molecular Biology, vol 1560. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-6788-9_18

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  • DOI: https://doi.org/10.1007/978-1-4939-6788-9_18

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  • Online ISBN: 978-1-4939-6788-9

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