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Histochemistry in Advanced Cytometry: From Fluorochromes to Mass Probes

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Histochemistry of Single Molecules

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2566))

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Abstract

For over half a century, fluorescence has been the milestone of most of the quantitative approaches in various fields from chemistry and biochemistry to microscopy. This latter also evolved into cytometry, thanks to the development of fluorescence techniques. The dyes of classical cytochemistry were replaced by fluorochromes, and the pioneer microphotometry was replaced by microfluorometry. The latter has great advantages in terms of simplicity, sensitivity, and accuracy. The extensive research and availability of new fluorochromes as well as the technological evolution contributed to the success of microfluorometry. The development of flow cytometry in the 1960s gave a giant boost to cell analysis and in particular to the clinical diagnostics. The synergy between flow cytometry and the subsequent development of monoclonal antibodies allowed the setup of multiparametric analytical panels that are today popular and irreplaceable in many clinical and research laboratories. Multiparametric analysis has required the application of an increasing number of fluorochromes, but their simultaneous use creates problems of mutual contamination, hence the need to develop new fluorescent probes. Semiconductor and nanotechnology research enabled the development of new probes called nanocrystals or quantum dots, which offered great advantages to the multiparametric analysis: in fact, thanks to their spectrofluorometric peculiarities, dozens of quantum dots may be simultaneously used without appreciable crosstalk between them. New analytical horizons in cytometry seem to be associated with a new concept of analysis that replaces fluorescence toward new markers with (non-radiative) isotopes of heavy metals. Thus, the mass flow cytometry was born, which seems to guarantee the simultaneous compensation-free analysis of up to 100 markers on a single sample aliquot.

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Authors and Affiliations

  1. Institute of Molecular Genetics – CNR (National Research Council), Pavia, Italy

    Giuliano Mazzini

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

    Giuliano Mazzini

  3. Department of Internal Medicine and Oncology, ASST Pavia and LIUCC University, Castellanza, Varese, 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

  3. Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, VERONA, Italy

    Manuela Malatesta

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Mazzini, G., Danova, M. (2023). Histochemistry in Advanced Cytometry: From Fluorochromes to Mass Probes. In: Pellicciari, C., Biggiogera, M., Malatesta, M. (eds) Histochemistry of Single Molecules. Methods in Molecular Biology, vol 2566. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2675-7_1

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  • DOI: https://doi.org/10.1007/978-1-0716-2675-7_1

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