Additional histological methods of stainig in preclinical studies

Y.A. Gushchin, 0000-0002-7656-991Х

Research-and-manufacturing company «Home оf Pharmacy», b. 245, 3, st. Zavodskaya, Kuzmolovskiy, Vsevolozhskiy district, Leningrad region, 188663, Russia



In clinical practice and in pharmacological studies, microscopic analysis is an integral part of the study of normal structure of tissues and pathologically altered organs. To prepare the material for histological examination, visualization is carried out by staining the tissue with dyes. The most common routine staining is hematoxylin and eosin, which is not enough to reveal the full picture of the process. Therefore, it is necessary to use a number of additional stains. Some of them can be used as routine instead of classical hematoxylin and eosin, for example, van Gieson stains or Mallory trichrome. Most stains are more specific, and they are used to identify specific structures or chemical compounds in cells and tissues. This allows you to obtain a significant amount of information, which facilitates an understanding the course of normal and pathological processes. Methods for the detection of mucopolysaccharides with Alcian blue are used in the study of the gastrointestinal tract and respiratory system. PAS staining is necessary in the diagnosis of accumulation diseases, some oncological processes and fungal infections. Fat-soluble dyes, primarily Sudan III and scharlach red are used in the study of dystrophic diseases and atherosclerosis. The Oil Red O dye is applicable for macroscopic assessment of the area of atherosclerotic lesions of the aorta. Congo red is the most popular dye for the amyloid detection. Specialized dyes for the diagnosis of myocardial damage were considered, such as (HBFP) (Haematoxylin-Basic Fuchsin-picric acid) and tetrasolium salts to visualize the area of damage to the heart muscle and brain. In this review, we examined several histological dyes, some features of their application, as well as mechanisms of the dye-substrate interaction in tissues. These staining methods can be used during histological work, and consider the possibility of their use, which will help to identify, as well as comprehensively study the pathological processes modeled in preclinical studies.

Full text avaliable in Russain only


  1. Black J. Microbiology: Principles and exploration (8th ed.). John Wiley & Sons, 2012: 68.
  2. Саркисов Д.С., Перов Ю.Л. Микроскопическая техника: Руководство для врачей и лаборантов. М.: Медицина, 1996: 542. [Sarkisov D.S., Perov Yu.L. Mikroskopicheskaya tekhnika: Rukovodstvo dlya vrachei i laborantov. M.: Meditsina, 1996: 542 (in Russ.)].
  3. Селиванов Е.В. Красители в биологии и медицине: Справочник. Барнаул: Азбука, 2003: 40 [Selivanov E.V. Krasiteli v biologii i meditsine: Spravochnik. – Barnaul: Azbuka, 2003: 40 (in Russ.)].
  4. Лилли Р. Патогистологическая техника и практическая гистохимия. М., 1969: 122 [Lilli R. Patogistologicheskaya tekhnika i prakticheskaya gistokhimiya. M., 1969: 122 (in Russ.)].
  5. Янин В.Л., Бондаренко О.М., Сазонова Н.А. Учебно-методическое пособие для аспирантов заочной формы обучения к практическим занятиям по дисциплине «Методы исследования в цитологии и гистологии». Учебно-методическое пособие. Ханты-Мансийск: БУ «Ханты-Мансийская государственная медицинская академия», 2015: 65. [Yanin V.L., Bondarenko O.M., Sazonova N.A. Uchebno-metodicheskoe posobie dlya aspirantov zaochnoi formy obucheniya k prakticheskim zanyatiyam po distsipline «Metody issledovaniya v tsitologii i gistologii». Uchebno-metodicheskoe posobie. Khanty-Mansiisk: BU «Khanty-Mansiiskaya gosudarstvennaya meditsinskaya akademiya», 2015: 65 (in Russ.)].
  6. Kiernan J.A. Dyes and other colorants in microtechnique and biomedical research. Coloration Technology 2006, 122 (1): 1–21. DOI: 10.1111/j.1478-4408.2006.00009.x
  7. Horobin R.W. Biological staining: mechanisms and theory. Biotech. Histochem, 2002; 77: 3–13. DOI: 10.1080/bih.
  8. Alturkistani H.A., Tashkandi F.M., Mohammedsaleh Z.M. Histological stains: A literature review and case study. Global Journal of Health Science. 2016; 8(3): 72–9. DOI: 10.5539/gjhs.v8n3p72
  9. Коржевский Д.Э. Применение гематоксилина в гистологической технике. Морфология, 2007, Т. 132, вып. 6: 77–82. [Korzhevskii D.E. Primenenie gematoksilina v gistologicheskoi tekhnike. Morfologiya, 2007, T. 132, vyp. 6: 77-82 (in Russ)]
  10. Семченко В.В., Барашкова С.А., Ноздрин В.Н., Артемьев В.Н. Гистологическая техника: учебное пособие. 3-е изд., доп. и перераб. Омск-Орел: Омская областная типография, 2011: 290. [Semchenko V.V., Barashkova S.A., Nozdrin V.N., Artem'ev V.N. Gistologicheskaya tekhnika: uchebnoe posobie. 3-e izd., dop. i pererab. Omsk-Orel: Omskaya oblastnaya tipografiya, 2011: 290 (in Russ.)].
  11. Prentø P: Van Gieson's picrofuchsin. The staining mechanisms for collagen and cytoplasm, and an examination of the dye diffusion rate model of differential staining. Histochemistry. 1993; 99: 163–74. DOI: 10.1007/bf00571877
  12. Коржевский Д.Э. Гиляров А.В. Основы гистологической техники. СПБ.: Спец.Лит. 2010: 95. [Korzhevskii D.E. Gilyarov A.V. Osnovy gistologicheskoi tekhniki. SPB, Spets.Lit. 2010: 95 (in Russ.)].
  13. Методы гистологической окраски. Каталог ООО «Биовитрум». М., Биовитрум, 2009:66 [Metody gistologicheskoi okraski. Katalog OOO «Biovitrum». M., Biovitrum, 2009:66 (in Russ)]
  14. Thomas F. McDonald, A. Bleak1ey Chandler Connective tissues in arterial and pulmonary disease. Springer-Verlag New York, 1981: 355. DOI 10.1007/978-1-4612-5967-1
  15. Horobin R.W. Structure-staining relationships in histochemistry and biological staining. I. Theoretical background and a general account of correlation of histochemical staining with the chemical structure of the reagents used. J Microsc. 1980; 119(3): 345–55. DOI: 10.1111/j.1365-2818.1980.tb04106.x
  16. Suvarna S. Kim, Layton Christopher, Bancroft John D. Bancroft's theory and practice of histological techniques. 7th edition. Philadelphia: Churchill Livingstone. 2008: 557.
  17. Пирс Э. Гистохимия. Издательство иностранной литературы, 1962:962. [Pirs E. Gistokhimiya/ Izdatel'stvo inostrannoi literatury, 1962:962. (in Russ.)].
  18. Bulmer D. Observations on Histological Methods Involving the use of Phosphotungstic and Phosphomolybdic Acids, with Particular Reference to Staining with Phosphotungstic Acid. Haematoxylin Journal of Cell Science. 1962; s3-103: 311–23.
  19. Луппа X. Основы гистохимии. М.: Мир, 1980: 343. [Luppa X. Osnovy gistokhimii. M.: Mir, 1980: 343 (in Russ)]
  20. Домницкий И.Ю., Салаутин В.В., Терентьев А.А. Секционный курс и методы патогистологических исследований: метод. пособие по выполнению лабораторных работ. Саратов: «Саратовский ГАУ». 2017: 50. [Domnitskii I.Yu., Salautin V.V., Terent'ev A.A. Sektsionnyi kurs i metody patogistologicheskikh issledovanii: metod. posobie po vypolneniyu laboratornykh rabot. FGBOU VO «Saratovskii GAU». Saratov, 2017: 50 (in Russ.)].
  21. Гущин Я.А., Мужикян А.А., Селезнева А.И., Макарова М.Н. Комплексная морфологическая оценка атеросклеротического повреждения аорт кроликов в эксперименте // Атеросклероз и дислипидемии. 2017; 1 (26): 50–9. [Gushchin Ya.A., Muzhikyan A.A., Selezneva A.I., Makarova M.N. Kompleksnaya morfologicheskaya otsenka ateroskleroticheskogo povrezhdeniya aort krolikov v eksperimente//Ateroskleroz i dislipidemii. 2017; 1 (26): 50–9 (in Russ)]
  22. Yakupova E. I., Bobyleva L.G, Vikhlyantsev I.M., Bobylev A.G. Congo Red and amyloids: history and relationship. Bioscience reports. 2019; 1: 39.
  23. Коржевский Д.Э. Морфологическая диагностика. Подготовка материала для гистологического исследования и электронной микроскопии. СПб.: СпецЛит, 2013: 127. [Korzhevskii D.E. Morfologicheskaya diagnostika. Podgotovka materiala dlya gistologicheskogo issledovaniya i elektronnoi mikroskopii. SPb.: SpetsLit, 2013: 127 (in Russ.)].
  24. Сапожников А.Г., Доросевич А.Е. Гистологическая и микроскопическая техника. Смоленск, 2000: 151–3 [Sapozhnikov A.G. , Dorosevich A.E. Gistologicheskaya i mikroskopicheskaya tekhnika. Smolensk, 2000: 151–3 (in Russ.)].
  25. Hatfield R.H., Mendelow A.D., Perry R.H., Alvarez L.M., Modha P. Triphenyltetrazolium chloride (TTC) as a marker for ischaemic changes in rat brain following permanent middle cerebral artery occlusion. Neuropathol Appl Neurobiol. 1991; 17 (1): 61–70.
  26. Bambhaniya A.B., Upadhyay M.C., Trangadia M.M., Mehta R.A., Gupta B.D., Chaudhari K.R. Role of Triphenyl Tetrazolium Chloride to detect early myocardial ischemia in sudden death cases. Journal of Research in Medical and Dental Science. 2015; Vol. 3: 54–9. DOI: 10.5455/jrmds.20153112

You may be interested