Bouin’s fluid will always be etched on my brain for a then commonplace but, with hindsight, foollhardy incident. During my final year at school I was a part-time laboratory steward (pay £2.14 shillings per week). One of my first jobs was to sort out the shelves at the back of the Advanced Biology Lab (now, along with the entire school, demolished). There was a 500 ml bottle half-full of Bouin’s fluid. The problem was that it had a ground-glass stopper. Even I knew that picric acid was an explosive and that anything containing picric acid should never come into contact with metal, must be kept dry and never put in anything with a ground-glass stopper. Friction or a sharp tap could detonate the dry material in the neck. I tried gently to remove the stopper. It was stuck. I had the wit not to try tapping the stopper on a bench to shift it because I could see dried picric acid in the neck. I then tried gentle heat from water heated with a bunsen burner (no piped hot water supply then) around the bottle. I grasped the stopper—there were no goggles or any other bits of safety equipment in the whole school—and pulled, without twisting. Out it came, but I don’t think I imagined hearing a small but definite ‘crack’. I now read that such episodes have to be dealt with by the bomb-disposal squad. I moved on to the next bottle. It was picric acid, obviously used in the past for the Bouin’s fluid. The contents were wet and it did have a rubber stopper.
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| Bouin's Fluid is available ready-mixed |
In the early and middle decades of the 20th century, the microscope was THE research tool in zoology—and many other -ology—laboratories. With the microscope came the paraphernalia to cut sections and stain the once-living material so that the various structures could be seen. Shelves in universities and schools were filled by a vast array of bottles containing various tissue fixatives, used to prevent shrinkage and decomposition, embedding materials, dyes, clearing agents, and mounting media for the thin sections. There was usually the evocative smell of Canada Balsam and xylol (then called xylene). Many of the mixtures of chemicals and dyes were named after the person who first used and described them during the golden age of histology. The obscure names on the faded labels of dusty bottles on equally dusty dark-stained wooden shelves added to the impression of alchemy rather than cutting-edge science.
Some of those named reagents have stood the test of time, while others have fallen by the wayside. The fixative of choice for the commonest staining method for mammalian tissues was and still is Bouin’s Fluid, a mixture of formaldehyde, acetic acid and picric acid. Having been aware of that fixative since the late 1950s, I had no idea who Bouin of his eponymous fluid was. Until last week that is when I saw a photograph taken at the First International Conference on Sex Hormones held in Paris in 1936. All of those present were well-known endocrinologists and I spotted those knew well, had met* or had heard of. And there was a Professor Pol Bouin of Strasbourg. Was he, I wondered, the Bouin of Bouin’s Fluid?
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| Pol Bouin circled From Zuckerman's biography, From Apes to Warlords |
Indeed he was. A bit of digging on Google soon turned up a biography. Pol André Bouin was born in Vendresse in northern France in 1870. While still a medical student in Nancy he became interested in morphology and was appointed preparateur d’histologie. He published the recipe of his new fixative in the same year that he qualified in medicine, 1897.
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| Pol Bouin from Parkes |
Bouin's rise in French science was meteoric, first in Nancy and then, after the First World War, in Strasbourg. His research covered a wide range of tissues and sub-cellular structures but he came to specialise in the reproductive organs and his fixative was devised for a study on seminiferous tubules. Much of his research and that of his collaborators in Strasbourg demonstrated the presence of putative hormones but preceded isolation and identification of the hormones themselves, prolactin and progesterone, for example. Similarly, his research in cytology, on the ergastoplasm (now the endoplasmic reticulum) was completely superseded once the electron microscope came on the scene.
Sir Alan Parkes—also on that photograph in Paris in 1936—wrote an obituary in Journal of Reproduction and Fertility. He began:
Pol Bouin, one of the pioneers of modern biology, and especially of the histological approach to physiological problems, died on February 5th, 1962, at the age of 92 at his family home in the Ardennes, to which he had retired finally in 1946. More than 20 years have elapsed since Bouin left active scientific work and time may have dimmed the bright light of his achievements for some of the younger generation. But in his own country, especially to his associates there, to the older generation elsewhere and to all with a sense of scientific history, Bouin ranks with F. H. A. Marshall and Ludwig Fraenkel as one of the small group of research workers who at the beginning of this century laid the foundations of our modern knowledge of the reproductive processes. Possibly even more significant than his personal original work was the influence he exerted directly and through his pupils. It may truly be said that Bouin founded a scientific dynasty which provided and continues to provide the major part of the great French contribution to our knowledge of the physiology of reproduction.
But Parkes made no mention at all of Bouin’s fluid!
*Solly Zuckerman, Ruth Deanesly, Alan Parkes, Idwal Rowlands, Frank Young
Bouin first reported the use of his fixative in this paper:
Etude sur l'évolution normale et l'involution du tube seminifere. I. Modifications régressives du processus spermatogénétique provoquées expérimentalement. II. Phénomènes cytologiques anormaux dans l'histogenèse et l'atrophie expérimentale du tube seminifere. Arch. Anat. micr. 1, 225-265. 1897
Ortiz-Hidalgo C. 1992. Pol André Bouin, MD (1870-1962). Bouin’s fixative and other contributions to medicine. Archives of Pathology and Laboratory Medicine 116, 882-884.
Parkes AS. 1963. Pol Bouin 1870-1962. A memoir. Journal of Reproduction and Fertility 5, 301-307.
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