doi:10.1017/S1551929514000583 18 www.microscopy-today.com • 2014 July Living Images from the Birth of Microscopy Brian J. Ford Gonville & Caius College, Cambridge University, Trinity Street, Cambridge CB2 1TA, UK brianjford@cardif.ac.uk Introduction Microscopes are probably used in more branches of scientific research than any comparable instrument, and the light microscope has become an instantly recognizable icon. Curiously, this single instrument is widely misrep- resented; the story of its development has for decades been misleading, and the general understanding of microscopy is weak. Worse, the media have done a great disservice to the instruments in general—they are poorly portrayed, wrongly interpreted, and both print and broadcast media have failed to do justice to the all-important role that the microscope has played in the advancement of our understanding. Ask yourself: when did you last see a worthwhile presen- tation of microscopic images on television or in a magazine? Te chances are it was a considerable time ago. Similarly, when did you see a successful flm of what the pioneering microscopists could see with their diminutive instruments? Tis is easier to answer: never. Tere have been occasional attempts to recreate the images from the dawn of microscopy, but none has been successful. Te standard works always emphasized the crude nature of early microscopes and the distorted and chromatic images that their inferior lenses could generate, and this has been perpetuated by recent television documentaries. Yet our research has radically reformed this accepted view. Even the minute, single lenses used by the pioneering microscopists can create images of startling clarity. As this research began, a number of micrographs were produced to show how good the results could be, and some of these have been honored with awards in Britain and America [1]. More recently, videomicrographs have been successfully obtained, and these provide, for the frst time, a living view of how the frst microscopists viewed their specimens. Te results belie the misleading accounts and demonstrate that the pioneer microscopists were observing nature with a clarity that is surprisingly close to what we might expect in today’s laboratory. This might seem to be an extreme statement, but consider: in so many fields of endeavor, the present- day version is much better than the first version, e.g., aircraft, automobiles, dwellings, computers, etc. Current devices make the earliest examples seem greatly inferior. Surprisingly, this is not the case with the conventional light microscope. Optical constraints impose a resolution limit of approximately 0.3 μ m, and it has been shown that the single-lens microscope hand-made by Antony van Leeuwenhoek and preserved at the University Museum for the History of Science at Utrecht, Netherlands, could resolve objects as fine as 0.7 μ m. The resolution of this early microscope was therefore within a factor of 3 of the best that a conventional light microscope could theoretically achieve. This may be unique in the history of science [2]. Materials and Methods Obtaining video images through these minute lenses is demanding, but not technically complex. The lens supplied on an SVGA webcam was removed and replaced with a customized bracket into which a small lens can be inserted. We have used original eighteenth- and nineteenth-century lenses from botanical and aquatic microscopes, as well as modern-day replica lenses. The best of these were plano- convex soda-glass lenses approximately 1 mm in diameter that were ground and polished by my colleague Es Reid of Cambridge University. The assembly is fitted onto a purpose-built bracket that provides an extension from the mechanical stage of an Olympus BH laboratory microscope. Use of the mechanical stage controls provides for movement of a specimen in the x-y directions; whereas, the coarse- and fine-focusing controls allow us to adjust for optimum image clarity. Illumination is provided by a single light- emitting diode mounted along the optical axis of the assembly and some 7–15 cm (about 3–6 inches) beneath the specimen. Actual early lenses ranged in magnification from 25 to 600 and allowed various experiments that recreated the observations recorded in the literature by pioneering microscopists. Recreations of Early Observations In our previous experiments, still micrographs were taken with the original microscope made by the pioneer of high-power microscopy, Antony van Leeuwenhoek of Delft, Netherlands, around 1690 and preserved at Utrecht. These modern photos taken through the original microscope showed that Leeuwenhoek’s own lenses could generate an image of astonishing quality. This was the microscope used by Bracegirdle in 1981 to image a blood smear. The results were disappointing, and no cells could be seen in the resulting image (Figure 1a). However, using a mount that fitted the diminutive microscope to a modern Olympus OM2n film camera allowed us to demonstrate how clear the image could be: a fresh blood smear could be observed in which, not only were the predominant erythrocytes clearly visible, but even the lobed nucleus of granulocyte could be resolved (Figure 1b). Thus, the resulting image taken with Leeuwenhoek’s original seventeenth- century microscope compares favorably with a present-day micrograph. In recent years, other attempts have been made to recreate observations made by pioneers from earlier centuries. The BBC program entitled Cell took the No. 2 lens from the microscope owned by Robert Brown in the 1820s and used it to reprise Brown’s observation of the https://doi.org/10.1017/S1551929514000583 Published online by Cambridge University Press