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Dmitri Leonidovich Romanowsky

Dmitri Leonidovich Romanowsky (sometimes spelled Dmitry and Romanowski, Russian: Дмитрий Леонидович Романовский;[1] 1861–1921) was a Russian physician who is best known for his invention of an eponymous histological stain called Romanowsky stain. It paved the way for the discovery and diagnosis of microscopic pathogens, such as malarial parasites,[2] and later developments of new histological stains that became fundamental to microbiology and physiology.[3]

Dmitri Leonidovich Romanowsky
Дмитрий Леонидович Романовский
Born1861
Pskov Governorate, Russia
Died1921(1921-00-00) (aged 59–60)
Kislovodsk
NationalityRussian
Alma materSt. Petersburg University
Known forRomanowsky stain
Scientific career
FieldsMedicine
Histochemistry
InstitutionsIvangorod military hospital
Revel local infirmary
Saint Petersburg Nikolaevsky military hospital
ThesisOn the question of parasitology and therapy of malaria (1891)

While working on his doctoral research, Romanowsky developed the first effective staining method for malarial parasite in 1890. Using a specific mixture of mouldy methylene blue and eosin, he found that malarial parasites could be distinctively identified from other blood cell and within the red blood cells. The chemical reaction of such staining is known in chemistry as "Romanowsky effect". The method became the gold standard in malaria detection by microscopy and general immunohistochemistry. British zoologist and science historian, Francis Edmund Gabriel Cox remarked the discovery as a serendipitous case that became "one of the most significant technical advances in the history of parasitology."[4]

Biography

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Romanowsky was born in 1861 in Pskov Governorate, Russia. He attended the 6th Saint Petersburg Gymnasium. In 1880, he enrolled at the St. Petersburg University. He enrolled for two courses: natural science (physics and mathematics) and medicine. He concentrated on medicine in 1882 for a preparatory course to the Military Medical Academy. He graduated with honors in 1886. On 30 November 1886, he was appointed as a junior resident of the Ivangorod military hospital. After one month, he was transferred to the Revel local infirmary as an associate doctor. In 1889, he was attached to the Saint Petersburg Nikolaevsky Military hospital. He initially worked at the clinical department, and from May 1890, he was the head of the eye department.[5] He obtained his medical degree in 1891 on the thesis "On the question of parasitology and therapy of malaria."[6][7]

Romanowsky died in 1921 in Kislovodsk in North Caucasus.[8]

Invention of histological stain

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Background

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Romanowsky's research for his medical degree in 1880s was mainly on the identification of malarial parasite (Plasmodium).[6] Until that time malarial infection was difficult to confirm as the parasites were hard to distinguish from blood cells or cell organelles. Pigmented blood cells were often linked to malarial infection, but the pigments are not always visible.[9] When French physician Charles Louis Alphonse Laveran discovered and described the malarial protozoan (later called Plasmodium falciparum) in 1880, it was not accepted as no protozoan had ever been seen in blood cells or associated with malaria.[10]

In 1871, German chemist Adolf von Baeyer synthesised a red dye called eosin (Greek word for "morning red"), which in 1876 was found to be useful for staining tissues.[11] Another German chemist Heinrich Caro synthesised a blue dye named methylene blue in 1876,[12] which was first used as a cell stain by Robert Koch. In 1882, using methylene blue Koch discovered the causative bacterium of tuberculosis, tubercle bacillus (now Mycobacterium tuberculosis).[13] The two stains remain among the fundamental stains used in general cell and tissue staining, as well as in clinical diagnosis.[11][14]

Romanowsky stain

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Romanowsky was the first to realise the differences in the staining abilities of eosin and methylene blue. The individual stains (monochromatic staining) were good only for general colouring of tissue or cell, but not for contrasting the different components.[15] By mixing specific amount of eosin and methylene blue, Romanowsky found that the mixture gave images of contrasting clarity that helped to visualise different parts and components of cells.[16] This mixture method, polychromatic staining or polychromy, with various modifications became the most efficient way of staining cells for identifying cellular components.[3][17] The chemical phenomenon by which a mixture of stains produces vibrant cell images is known as "Romanowsky effect".[15][18]

In December 1890, Romanowsky published his invention as a preliminary report of his major work for his doctoral thesis in the journal Vrach as "On the question of the structure of malaria parasites" (as translated in English).[19][20] Incorrectly, it is more often recorded in books and journals that Romanowsky published his findings in 1891,[8][21][22][23] which led to a controversy on priority that Ernst Malachowsky independently developed the technique as the latter published his research in August 1891.[20]

Romanowsky discovered that instead of fresh methylene blue, an aged and mouldy solution gave the best result, while eosin should be free of any contamination.[8][24] He described:

For staining [blood sample having malarial infection] the following mixture is used, as discovered by me, which is best when freshly prepared: 2 volumes of a filtered saturated aqueous solution of methylene blue plus 5 volumes of a 1% aqueous eosin solution... In my preparations I always obtain the following picture. Red cells are stained in a pink color. Cytoplasm in eosinophils is saturated-pink, whilst that in the malaria parasite and lymphocytes is light blue. Blood platelets and the nuclei of white cell are dark-violet, whilst the nuclei of malaria parasites are purple-violet. The cytoplasm of leukocytes is pale-violet, with transitional colors between the light blue protoplasm of lymphocytes to violet leukocytes. Within red cells the malaria parasite may be hardly noticeable or may occupy the whole cell. In any event, the violet nucleus, surrounded by a colorless rim, is always clearly distinguishable.[20]

Romanowsky gave an elaborate description of the new technique in his thesis submitted in June 1891.[8] The staining method remains the "gold standard" for visualising blood samples, especially for malarial infection,[2] and in immunohistochemical studies.[24]

References

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  1. ^ Veer, M. v.; Haferlach, T. (2014). "Should clinical hematologists put their microscopes on eBay?". Haematologica. 99 (10): 1533–1534. doi:10.3324/haematol.2014.114710. PMC 4181246. PMID 25271310.
  2. ^ a b Fleischer, Bernhard (2004). "Editorial: 100 years ago: Giemsa's solution for staining of plasmodia". Tropical Medicine and International Health. 9 (7): 755–756. doi:10.1111/j.1365-3156.2004.01278.x. PMID 15228484.
  3. ^ a b Wittekind, D. H. (1983). "On the nature of Romanowsky--Giemsa staining and its significance for cytochemistry and histochemistry: an overall view". The Histochemical Journal. 15 (10): 1029–1047. doi:10.1007/BF01002498. ISSN 0018-2214. PMID 6196323. S2CID 23896062.
  4. ^ Cox, Francis Eg (2010). "History of the discovery of the malaria parasites and their vectors". Parasites & Vectors. 3 (1): 5. doi:10.1186/1756-3305-3-5. PMC 2825508. PMID 20205846. S2CID 1641027.
  5. ^ "IN MEMORIAM OF RUSSIAN DOCTORS - ROMANOWSKY DMITRY LEONIDOVICH AND CHENZINSKY CHESLAV IVANOVICH - IT IS DEVOTED". EMCO LTD. Retrieved 29 February 2016.
  6. ^ a b Bynum, William F.; Overy, Caroline (1998). The Beast in the Mosquito: The Correspondence of Ronald Ross and Patrick Manson. Rodopi. p. 418. ISBN 978-90-420-0721-5.
  7. ^ Grove, David I (2014). Tapeworms, Lice, and Prions: A Compendium of Unpleasant Infections. Oxford: Oxford University Press, Incorporated. pp. 124–126. ISBN 978-0-1996410-24.
  8. ^ a b c d Krafts, K. P.; Hempelmann, E.; Oleksyn, B. J. (2011). "The color purple: from royalty to laboratory, with apologies to Malachowski". Biotechnic & Histochemistry. 86 (1): 7–35. doi:10.3109/10520295.2010.515490. ISSN 1473-7760. PMID 21235291. S2CID 19829220.
  9. ^ Manson-Bahr, P. (1961). "The malaria story". Proceedings of the Royal Society of Medicine. 54 (2): 91–100. doi:10.1177/003591576105400202. PMC 1870294. PMID 13766295.
  10. ^ Lalchhandama, K (2014). "The making of modern malariology: from miasma to mosquito-malaria theory" (PDF). Science Vision. 14 (1): 3–17. Archived from the original (PDF) on 2014-04-27.
  11. ^ a b King, D. F.; King, L. A. (1986). "A brief historical note on staining by hematoxylin and eosin". The American Journal of Dermatopathology. 8 (2): 168. doi:10.1097/00000372-198604000-00013. ISSN 0193-1091. PMID 2424331.
  12. ^ Cooksey, C. J. (2017). "Quirks of dye nomenclature. 8. Methylene blue, azure and violet". Biotechnic & Histochemistry. 92 (5): 347–356. doi:10.1080/10520295.2017.1315775. ISSN 1473-7760. PMID 28598697. S2CID 46746062.
  13. ^ Sakula, A. (1983). "Robert koch: centenary of the discovery of the tubercle bacillus, 1882". The Canadian Veterinary Journal. 24 (4): 127–131. ISSN 0008-5286. PMC 1790283. PMID 17422248.
  14. ^ Oz, Murat; Lorke, Dietrich E.; Hasan, Mohammed; Petroianu, George A. (2011). "Cellular and molecular actions of Methylene Blue in the nervous system". Medicinal Research Reviews. 31 (1): 93–117. doi:10.1002/med.20177. PMC 3005530. PMID 19760660.
  15. ^ a b Krafts, K. P.; Pambuccian, S. E. (2011). "Romanowsky staining in cytopathology: history, advantages and limitations". Biotechnic & Histochemistry. 86 (2): 82–93. doi:10.3109/10520295.2010.515492. ISSN 1473-7760. PMID 21395493. S2CID 5168332.
  16. ^ Horobin, R. W. (2011). "How Romanowsky stains work and why they remain valuable - including a proposed universal Romanowsky staining mechanism and a rational troubleshooting scheme". Biotechnic & Histochemistry. 86 (1): 36–51. doi:10.3109/10520295.2010.515491. ISSN 1473-7760. PMID 21235292. S2CID 207513741.
  17. ^ Jörundsson, Einar; Lumsden, John H.; Jacobs, Robert M. (1999). "Rapid staining techniques in cytopathology: a review and comparison of modified protocols for hematoxylin and eosin, Papanicolaou and Romanowsky stains". Veterinary Clinical Pathology. 28 (3): 100–108. doi:10.1111/j.1939-165x.1999.tb01057.x. ISSN 1939-165X. PMID 12075519.
  18. ^ Bianco, P.; Ponzi, A.; Bonucci, E. (1984). "Basic and 'special' stains for plastic sections in bone marrow histopathology, with special reference to May-Grünwald Giemsa and enzyme histochemistry". Basic and Applied Histochemistry. 28 (3): 265–279. ISSN 0391-7258. PMID 6083774.
  19. ^ Romanowsky, D. L. (1890). "To a question of a structure of malaria parasites" (PDF). Vrach. 11: 1171–1173.
  20. ^ a b c Bezrukov, A. V. (2017). "Romanowsky staining, the Romanowsky effect and thoughts on the question of scientific priority". Biotechnic & Histochemistry. 92 (1): 29–35. doi:10.1080/10520295.2016.1250285. ISSN 1473-7760. PMID 28098484. S2CID 37401579.
  21. ^ Journal of Applied Microscopy. Bausch & Lomb Optical Company. 1903. p. 2229.
  22. ^ Gill, Gary (2012). Cytopreparation: Principles & Practice. Springer Science & Business Media. p. 218. ISBN 978-1-4614-4933-1.
  23. ^ Menzies, D. W. (1962). "Romanowsky-type staining: effect of adding acid dyes, particularly chromotrope 2R". Stain Technology. 37: 45–48. doi:10.3109/10520296209114570. ISSN 0038-9153. PMID 14472722.
  24. ^ a b Kalinin, Valeriy; Padnya, Pavel; Stoikov, Ivan (2023). "Romanowsky staining: history, recent advances and future prospects from a chemistry perspective". Biotechnic & Histochemistry. 99 (1): 1–20. doi:10.1080/10520295.2023.2273860. ISSN 1473-7760. PMID 37929609. S2CID 265032156.
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