https://doi.org/10.4081/ejh.2024.3989
The Feulgen reaction at the electron microscopy level
Submitted: 12 February 2024
Accepted: 14 February 2024
Published: 23 February 2024
Accepted: 14 February 2024
Abstract Views: 460
PDF: 213
HTML: 3
HTML: 3
Publisher's note
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
All claims expressed in this article are solely those of the authors and do not necessarily represent those of their affiliated organizations, or those of the publisher, the editors and the reviewers. Any product that may be evaluated in this article or claim that may be made by its manufacturer is not guaranteed or endorsed by the publisher.
Authors
The Feulgen reaction has been the first specific method for detecting DNA available at light microscopy since 1924. However, a similar specific method was proposed for electron microscopy only 50 years later. Here, we discuss the problems encountered in finding the electrondense reagent capable of taking advantage of the extremely high resolution offered by electron microscopy as well as some applications of the method.
Hayat H. Stains and cytochemical methods. New York: Plenum Press; 1993. pp. 254-60.
Feulgen R, Rossenbeck H. [Mikroskopisch-chemischer Nachweis einer Nukleinsäure von Typus der Thymonukleinsäure und die darauf beruhende selektive Färbung von Zellkernen in mikroskopischen Präparaten].[Article in German]. Hoppe-Seylers Z Physiol Chem 1924;135:203-48.
DOI: https://doi.org/10.1515/bchm2.1924.135.5-6.203
Mazzini G. The Feulgen reaction: from pink-magenta to fluorescent rainbow at the MaffoVialli’s School of Histochemistry. Eur J Histochem 2024;68:3971.
DOI: https://doi.org/10.4081/ejh.2024.3971
Gautier A. Ultrastructural localization of DNA in ultrathin tissue sections. Int Rev Cytol 1976;44:113-91.
DOI: https://doi.org/10.1016/S0074-7696(08)61649-6
Moyne G. Methods in ultrastructural cytochemistry of the cell nucleus. Prog Histochem Cytochem 1980;13:1-72.
DOI: https://doi.org/10.1016/S0079-6336(80)80008-8
Bendayan M, Puvion E. Ultrastructural localization of nucleic acids through several cytochemical techniques on osmium-fixed tissues: comparative evaluation of the different labelings. J Histochem Cytochem 1984;32:1185-91.
DOI: https://doi.org/10.1177/32.11.6208238
Kasten FH. The chemistry of Schiff's reagent. Int Rev Cytol 1960;10:1-100.
Feulgen R, Volt K. [Über den Mechanismus der Nuclealfärbung. I Mitteilung. Über den Nachweis der reduzierenden Gruppen in den Kernen partiell hydrolysierter mikroskopischer Präparate].[Article in German]. Biol Chem 1924;135:249-52.
DOI: https://doi.org/10.1515/bchm2.1924.135.5-6.249
Bretschneider LH. An electron microscopical study of bull sperm. Proc K Ned Akad Wet 1949;52:301-9.
Peters D. Electron microscopic studies on the localization of deoxyribonucleic acid inside of DNA viruses. In: Uyeda R (ed.) Electron microscopy, vol 2. Tokyo: Maruzen; 1966. pp. 195-6.
Moyne G. Feulgen-derived techniques for electron microscopical cytochemistry of DNA. J Ultrastruct Res 1973;45:102-23.
DOI: https://doi.org/10.1016/S0022-5320(73)90036-1
Cogliati R, Gautier A. [Mise en evidence de l'ADN et des polysaccharides à l'aide d'un nouveau réactif "de type Schiff"].[Article in French]. C R Acad Sci 1973;276:3041-4.
Gautier A, Cogliati R, Schreyer M, Fakau J. Ultrastructural cytochemistry: a new specific stain for DNA and polysaccharides. Experientia 1973;29:771.
Olins AL, Moyer BA, Kim SH, Allison DP. Synthesis of a more stable osmium ammine electron dense stain. J Histochem Cytochem 1989;37:395-8.
DOI: https://doi.org/10.1177/37.3.2465337
Mikhaylova VT, Markov DV. An alternative method for preparation of Schiff-like reagent from osmium-ammine complex for selective staining of DNA on thin Lowicryl sections. J Histochem Cytochem 1994;42:1643-9.
DOI: https://doi.org/10.1177/42.12.7983365
Vázquez-Nin GH, Biggiogera M, Echeverría OM. Activation of osmium ammine by SO2-generating chemicals for EM Feulgen-type staining of DNA. Eur J Histochem 1995;39:101-6.
Masiello I, Biggiogera M. Osmium ammine for staining DNA in electron microscopy. Methods Mol Biol 2017;1560:261-7.
DOI: https://doi.org/10.1007/978-1-4939-6788-9_19
Derenzini M, Farabegoli F. Selective staining of nucleic acids by osmium ammine complex in thin sections from Lowicryl-embedded samples. J Histochem Cytochem 1990;28:1495-501.
DOI: https://doi.org/10.1177/38.10.2205645
Olins AL, Olins DE, Bazett-Jones DP. Osmium ammine-B and electron spectroscopic imaging of ribonucleoproteins: correlation of stain and phosphorus. Biol Cell 1996;87:143-7.
DOI: https://doi.org/10.1111/j.1768-322X.1996.tb00976.x
Olins AL, Olins DE, Olman V, Levy HA, Bazett-Jones DP. Modeling the 3-D RNA distribution in the Balbiani ring granule. Chromosoma 1994;103:302-10.
DOI: https://doi.org/10.1007/s004120050037
Derenzini M, Hernandez-Verdun D, Bouteille M. Visualization of a repeating subunit organization in rat hepatocyte chromatin fixed in situ. J Cell Sci 1983;61:137-49.
DOI: https://doi.org/10.1242/jcs.61.1.137
Derenzini M, Viron A, Puvion-Dutilleul F. The Feulgen-like osmium-ammine reaction as a tool to investigate chromatin structure in thin sections. J Ultrastruct Res 1982;80:133-47.
DOI: https://doi.org/10.1016/S0022-5320(82)90013-2
Hernandez-Verdun D, Derenzini M. Non-nucleosomal configuration of chromatin in nucleolar organizer regions of metaphase chromosomes in situ. Eur J Cell Biol 1983;31:360-5.
Hernandez-Verdun D, Derenzini M, Bouteille M. The morphological relationship in electron microscopy between NOR-silver proteins and intranucleolar chromatin. Chromosoma 1982;85:461-73.
DOI: https://doi.org/10.1007/BF00327343
Puvion E, Bernhard W. Ribonucleoprotein components in liver cell nuclei as visualized by cryoultramicrotomy. J Cell Biol 1975;67:200-14.
DOI: https://doi.org/10.1083/jcb.67.1.200
Biggiogera M. Chromatin arrangement in mouse sperm nuclei: an ultrastructural cytochemical study. Mol Reprod Dev 1989;1:91-7.
DOI: https://doi.org/10.1002/mrd.1080010203
Courtens JL, Biggiogera M, Fakan S. A cytochemical and immunocytochemical study of DNA distribution in spermatid nuclei of mouse, rabbit, and bull. Cell Tissue Res 1991;265:517-25.
DOI: https://doi.org/10.1007/BF00340875
Courtens JL, Biggiogera M, Fakan S. Distribution of DNA, nuclear micro-heterogeneities and compaction of the chromatin in rabbit epididymal spermatozoa. Ultrastructural evaluation of the Feulgen-like technique using osmium ammine. Reprod Nutr Dev 1994;34:261-72.
DOI: https://doi.org/10.1051/rnd:19940308
Boutinard Rouelle-Rossier V, Biggiogera M. Electron spectroscopic imaging and X-ray microanalysis of phosphorus in mouse sperm chromatin. Eur J Histochem 1992;36:197-204.
Fakan S, Odartchenko N. Ultrastructural organization of the cell nucleus in early mouse embryos. Biol Cell 1980;37:211-8.
Biggiogera M, Courtens JL, Derenzini M, Fakan S, Hernandez-Verdun D, Risueno MC, et al. Osmium ammine: review of current applications to visualize DNA in electron microscopy. Biol Cell 1996;87:121-32.
DOI: https://doi.org/10.1111/j.1768-322X.1996.tb00974.x
Puvion-Dutilleul F, Leduc EH, Puvion E. The osmium ammine-SO2 staining method for studying the in situ configuration of viral genomes in ultrathin sections of DNA virus infected cells. Biol Cell 1996;87:133-41.
DOI: https://doi.org/10.1016/S0248-4900(97)89270-2
Derenzini M, Olins AL, Olins DE. Chromatin structure in situ: the contribution of DNA ultrastructural cytochemistry. Eur J Histochem 2014;58:2307.
DOI: https://doi.org/10.4081/ejh.2014.2307
Ohwada Y, Eguchi M. Ultrastructural investigation of DNA in megakaryoblastic leukemia by using osmium-ammine-B: comparison with several types of leukemia. Med Oncol Tumor Pharmacothe. 1993;10:117-24.
DOI: https://doi.org/10.1007/BF02987778
el-Alfy M, Liu DF, Leblond CP. DNA changes involved in the formation of metaphase chromosomes, as observed in mouse duodenal crypt cells stained by osmium-ammine. I. New structures arise during the S phase and condense at prophase into “chromomeres” which fuse at prometaphase into mitotic chromosomes. Anat Rec 1995;242:433-48.
DOI: https://doi.org/10.1002/ar.1092420402
Liu DF, el-Alfy M, Leblond CP. DNA changes involved in the formation of metaphase chromosomes, as observed in mouse duodenal crypt cells stained by osmium-ammine. II. Tracing nascent DNA by bromodeoxyuridine into structures arising during the S phase. Anat Rec 1995;242:449-61.
DOI: https://doi.org/10.1002/ar.1092420403
Biggiogera M, Malatesta M, Abolhassani-Dadras S, Amalric F, Rothblum LI, Fakan S. Revealing the unseen: the organizer region of the nucleolus. J Cell Sci 2001;114:3199-205.
DOI: https://doi.org/10.1242/jcs.114.17.3199
von Schack ML, Fakan S. Retention of glycogen in cryosubstituted mouse liver. Histochemistry 1994;102:451-5.
DOI: https://doi.org/10.1007/BF00269577
How to Cite
Biggiogera, M. (2024). The Feulgen reaction at the electron microscopy level. European Journal of Histochemistry, 68(1). https://doi.org/10.4081/ejh.2024.3989
Copyright (c) 2024 The Author(s)
This work is licensed under a Creative Commons Attribution-NonCommercial 4.0 International License.
PAGEPress has chosen to apply the Creative Commons Attribution NonCommercial 4.0 International License (CC BY-NC 4.0) to all manuscripts to be published.
Similar Articles
- M. Malatesta, C. Zancanaro, M. Costanzo, B. Cisterna, C. Pellicciari, Simultaneous ultrastructural analysis of fluorochrome-photoconverted diaminobenzidine and gold immunolabeling in cultured cells , European Journal of Histochemistry: Vol. 57 No. 3 (2013)
- S. Grecchi, M. Malatesta, Visualizing endocytotic pathways at transmission electron microscopy via diaminobenzidine photo-oxidation by a fluorescent cell-membrane dye , European Journal of Histochemistry: Vol. 58 No. 4 (2014)
- M. Malatesta, M. Giagnacovo, M. Costanzo, B. Conti, I. Genta, R. Dorati, V. Galimberti, M. Biggiogera, C. Zancanaro, Diaminobenzidine photoconversion is a suitable tool for tracking the intracellular location of fluorescently labelled nanoparticles at transmission electron microscopy , European Journal of Histochemistry: Vol. 56 No. 2 (2012)
- M. Derenzini, A. L. Olins, D. E. Olins, Chromatin structure in situ: the contribution of DNA ultrastructural cytochemistry , European Journal of Histochemistry: Vol. 58 No. 1 (2014)
- M. Costanzo, F. Carton, A. Marengo, G. Berlier, B. Stella, S. Arpicco, M. Malatesta, Fluorescence and electron microscopy to visualize the intracellular fate of nanoparticles for drug delivery , European Journal of Histochemistry: Vol. 60 No. 2 (2016)
- D. Martini, A. Trirè, L. Breschi, A. Mazzoni, G. Teti, M. Falconi, A. Ruggeri Jr, Dentin matrix protein 1 and dentin sialophosphoprotein in human sound and carious teeth: an immunohistochemical and colorimetric assay , European Journal of Histochemistry: Vol. 57 No. 4 (2013)
- M. Scimeca, A. Orlandi, I. Terrenato, S. Bischetti, E. Bonanno, Assessment of metal contaminants in non-small cell lung cancer by EDX microanalysis , European Journal of Histochemistry: Vol. 58 No. 3 (2014)
- M. Miko, L. Danisovic, A. Majidi, I. Varga, Ultrastructural analysis of different human mesenchymal stem cells after in vitro expansion: a technical review , European Journal of Histochemistry: Vol. 59 No. 4 (2015)
- Simona Polakovičová, Mária Csӧbӧnyeiová, Barbora Filova, Miroslav Borovský, Ladislav Maršík, Alena Kvasilová, Štefan Polák, Merkel-like cell distribution in the epithelium of the human vagina. An immunohistochemical and TEM study , European Journal of Histochemistry: Vol. 62 No. 1 (2018)
- W. Meyer, J. Kacza, I. N. Hornickel, B. Schoennagel, Immunolocalization of succinate dehydrogenase in the esophagus epithelium of domesticated mammals , European Journal of Histochemistry: Vol. 57 No. 2 (2013)
You may also start an advanced similarity search for this article.
Publication Facts
Metric
This article
Other articles
Peer reviewers
0
2.4
Reviewer profiles N/A
Author statements
Author statements
This article
Other articles
Data availability
N/A
16%
External funding
N/A
32%
Competing interests
N/A
11%
Metric
This journal
Other journals
Articles accepted
57%
33%
Days to publication
10
145
- Editor & editorial board
- profiles
- Academic society
- N/A
- Publisher
- PAGEPress Publications, Pavia, Italy
To learn about these publication facts, click 
PF is maintained by the Public Knowledge Project
