Optical microscope

1449: 1249: 329: 1067: 966: 805: 1181: 1105: 532:(1632–1724) is credited with bringing the microscope to the attention of biologists, even though simple magnifying lenses were already being produced in the 16th century. Van Leeuwenhoek's home-made microscopes were simple microscopes, with a single very small, yet strong lens. They were awkward in use, but enabled van Leeuwenhoek to see detailed images. It took about 150 years of optical development before the compound microscope was able to provide the same quality image as van Leeuwenhoek's simple microscopes, due to difficulties in configuring multiple lenses. In the 1850s, 741:, or ocular lens, is a cylinder containing two or more lenses; its function is to bring the image into focus for the eye. The eyepiece is inserted into the top end of the body tube. Eyepieces are interchangeable and many different eyepieces can be inserted with different degrees of magnification. Typical magnification values for eyepieces include 5×, 10× (the most common), 15× and 20×. In some high performance microscopes, the optical configuration of the objective lens and eyepiece are matched to give the best possible optical performance. This occurs most commonly with 1048: 1086: 22: 666: 1501: 185: 3278: 128: 3379: 518: 452:, invented the compound microscope and/or the telescope as early as 1590. Johannes' testimony, which some claim is dubious, pushes the invention date so far back that Zacharias would have been a child at the time, leading to speculation that, for Johannes' claim to be true, the compound microscope would have to have been invented by Johannes' grandfather, Hans Martens. Another claim is that Janssen's competitor, 3391: 204:) that gives the viewer an enlarged inverted virtual image of the object (image 2). The use of a compound objective/eyepiece combination allows for much higher magnification. Common compound microscopes often feature exchangeable objective lenses, allowing the user to quickly adjust the magnification. A compound microscope also enables more advanced illumination setups, such as 1512:

molecule produces a diffraction-limited spot of light in the image, and the centre of each of these spots corresponds to the location of the molecule. As the number of fluorescing molecules is low the spots of light are unlikely to overlap and therefore can be placed accurately. This process is then repeated many times to generate the image.

1484:, Alexa dyes, Atto dyes, Cy2/Cy3 and fluorescein molecules can be used for localization microscopy, provided certain photo-physical conditions are present. Using this so-called SPDMphymod (physically modifiable fluorophores) technology a single laser wavelength of suitable intensity is sufficient for nanoimaging. 873:

At magnifications higher than 100× moving a slide by hand is not practical. A mechanical stage, typical of medium and higher priced microscopes, allows tiny movements of the slide via control knobs that reposition the sample/slide as desired. If a microscope did not originally have a mechanical stage

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The frame provides a mounting point for various microscope controls. Normally this will include controls for focusing, typically a large knurled wheel to adjust coarse focus, together with a smaller knurled wheel to control fine focus. Other features may be lamp controls and/or controls for adjusting

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is sometimes cited as a compound microscope inventor. After 1610, he found that he could close focus his telescope to view small objects, such as flies, close up and/or could look through the wrong end in reverse to magnify small objects. The only drawback was that his 2 foot long telescope had to be

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of a microscope is taken as the ability to distinguish between two closely spaced Airy disks (or, in other words the ability of the microscope to reveal adjacent structural detail as distinct and separate). It is these impacts of diffraction that limit the ability to resolve fine details. The extent

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or water and a matched cover slip between the objective lens and the sample. The refractive index of the index-matching material is higher than air allowing the objective lens to have a larger numerical aperture (greater than 1) so that the light is transmitted from the specimen to the outer face of

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and the objective lens. For example a 10x eyepiece magnification and a 100x objective lens magnification gives a total magnification of 1,000×. Modified environments such as the use of oil or ultraviolet light can increase the resolution and allow for resolved details at magnifications larger than

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Focusing starts at lower magnification in order to center the specimen by the user on the stage. Moving to a higher magnification requires the stage to be moved higher vertically for re-focus at the higher magnification and may also require slight horizontal specimen position adjustment. Horizontal

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Adjustment knobs move the stage up and down with separate adjustment for coarse and fine focusing. The same controls enable the microscope to adjust to specimens of different thickness. In older designs of microscopes, the focus adjustment wheels move the microscope tube up or down relative to the

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is a simple example of how higher resolution surpassing the diffraction limit is possible, but it has major limitations. STED is a fluorescence microscopy technique which uses a combination of light pulses to induce fluorescence in a small sub-population of fluorescent molecules in a sample. Each

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that collect light from the sample. The objective is usually in a cylinder housing containing a glass single or multi-element compound lens. Typically there will be around three objective lenses screwed into a circular nose piece which may be rotated to select the required objective lens. These

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lenses with different magnification are usually provided mounted on a turret, allowing them to be rotated into place and providing an ability to zoom-in. The maximum magnification power of optical microscopes is typically limited to around 1000x because of the limited resolving power of visible

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Multiple techniques are available for reaching resolutions higher than the transmitted light limit described above. Holographic techniques, as described by Courjon and Bulabois in 1979, are also capable of breaking this resolution limit, although resolution was restricted in their experimental

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Bradl, Joachim (1996). "Comparative study of three-dimensional localization accuracy in conventional, confocal laser scanning and axial tomographic fluorescence light microscopy". In Bigio, Irving J; Grundfest, Warren S; Schneckenburger, Herbert; Svanberg, Katarina; Viallet, Pierre M (eds.).

1588:). The specimen chambers needed for all such instruments also limits sample size, and sample manipulation is more difficult. Color cannot be seen in images made by these methods, so some information is lost. They are however, essential when investigating molecular or atomic effects, such as 835:

the objective lens with minimal refraction. Numerical apertures as high as 1.6 can be achieved. The larger numerical aperture allows collection of more light making detailed observation of smaller details possible. An oil immersion lens usually has a magnification of 40 to 100×.

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to generate magnified images of small objects. Optical microscopes are the oldest design of microscope and were possibly invented in their present compound form in the 17th century. Basic optical microscopes can be very simple, although many complex designs aim to improve

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All modern optical microscopes designed for viewing samples by transmitted light share the same basic components of the light path. In addition, the vast majority of microscopes have the same 'structural' components (numbered below according to the image on the right):

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Modern microscopes allow more than just observation of transmitted light image of a sample; there are many techniques which can be used to extract other kinds of data. Most of these require additional equipment in addition to a basic compound microscope.

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Aspden, Reuben S.; Gemmell, Nathan R.; Morris, Peter A.; Tasca, Daniel S.; Mertens, Lena; Tanner, Michael G.; Kirkwood, Robert A.; Ruggeri, Alessandro; Tosi, Alberto; Boyd, Robert W.; Buller, Gerald S.; Hadfield, Robert H.; Padgett, Miles J. (2015).

353:. Microscopes can also be partly or wholly computer-controlled with various levels of automation. Digital microscopy allows greater analysis of a microscope image, for example, measurements of distances and areas and quantitation of a fluorescent or 1580:

STM and AFM are scanning probe techniques using a small probe which is scanned over the sample surface. Resolution in these cases is limited by the size of the probe; micromachining techniques can produce probes with tip radii of 5–10 nm.

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The use of electrons and X-rays in place of light allows much higher resolution – the wavelength of the radiation is shorter so the diffraction limit is lower. To make the short-wavelength probe non-destructive, the atomic beam imaging system

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The whole of the optical assembly is traditionally attached to a rigid arm, which in turn is attached to a robust U-shaped foot to provide the necessary rigidity. The arm angle may be adjustable to allow the viewing angle to be adjusted.

564:. This method of sample illumination gives rise to extremely even lighting and overcomes many limitations of older techniques of sample illumination. Before development of Köhler illumination the image of the light source, for example a 139:

of a single lens or group of lenses for magnification. A compound microscope uses a system of lenses (one set enlarging the image produced by another) to achieve a much higher magnification of an object. The vast majority of modern

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It is important to note that higher frequency waves have limited interaction with matter, for example soft tissues are relatively transparent to X-rays resulting in distinct sources of contrast and different target applications.

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for light microscopy. "Optically isolated" means that at a given point in time, only a single particle/molecule within a region of a size determined by conventional optical resolution (typically approx. 200–250 nm

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The stage is a platform below the objective lens which supports the specimen being viewed. In the center of the stage is a hole through which light passes to illuminate the specimen. The stage usually has arms to hold

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of the Max Planck Institute for Biophysical Chemistry was awarded the 10th German Future Prize in 2006 and Nobel Prize for Chemistry in 2014 for his development of the STED microscope and associated methodologies.

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Journal of the Royal Microscopical Society, Containing Its Transactions and Proceedings and a Summary of Current Researches Relating to Zoology and Botany (Principally Invertebrata and Cryptogamia), Microscopy,

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While basic microscope technology and optics have been available for over 400 years it is much more recently that techniques in sample illumination were developed to generate the high quality images seen today.

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Lemmer, P.; Gunkel, M.; Baddeley, D.; Kaufmann, R.; Urich, A.; Weiland, Y.; Reymann, J.; Müller, P.; Hausmann, M.; Cremer, C. (2008). "SPDM: light microscopy with single-molecule resolution at the nanoscale".

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port to show the images directly on the monitor. They offer modest magnifications (up to about 200×) without the need to use eyepieces and at a very low cost. High-power illumination is usually provided by an

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Kaufmann, R; Müller, P; Hildenbrand, G; Hausmann, M; Cremer, C; et al. (2011). "Analysis of Her2/neu membrane protein clusters in different types of breast cancer cells using localization microscopy".

1104: 259:, whose design usually includes a polarizing filter, rotating stage, and gypsum plate to facilitate the study of minerals or other crystalline materials whose optical properties can vary with orientation. 1066: 400:

to photon-sparse microscopy, the sample is illuminated with infrared photons, each spatially correlated with an entangled partner in the visible band for efficient imaging by a photon-counting camera.

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are simple single-lens microscopes. Compound microscopes can be further divided into a variety of other types of microscopes, which differ in their optical configurations, cost, and intended purposes.

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Due to the difficulty in preparing specimens and mounting them on slides, for children it is best to begin with prepared slides that are centered and focus easily regardless of the focus level used.

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corrected, and therefore a huge step forward in microscope development. The Huygens ocular is still being produced to this day, but suffers from a small field size, and other minor disadvantages.

2064:, p. 28) makes it unlikely he invented it in 1590 and the claim of invention is based on the testimony of Zacharias Janssen's son, Johannes Zachariassen, who may have fabricated the whole story ( 1492:

3D super resolution microscopy with standard fluorescent dyes can be achieved by combination of localization microscopy for standard fluorescent dyes SPDMphymod and structured illumination SMI.

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Robert D. Huerta, Giants of Delft: Johannes Vermeer and the Natural Philosophers : the Parallel Search for Knowledge During the Age of Discovery, Bucknell University Press - 2003, page 126

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Note: stories vary, including Zacharias Janssen had the help of his father Hans Martens (or sometimes said to have been built entirely by his father). Zacharias' probable birth date of 1585 (

456:(who applied for the first telescope patent in 1608) also invented the compound microscope. Other historians point to the Dutch innovator Cornelis Drebbel with his 1621 compound microscope. 2874: 1222:

associated with long workdays at a microscopy station. In certain applications, long-working-distance or long-focus microscopes are beneficial. An item may need to be examined behind a

3071:"Metallographic and Materialographic Specimen Preparation, Light Microscopy, Image Analysis and Hardness Testing", Kay Geels in collaboration with Struers A/S, ASTM International 2006. 1334: 216:

There are many variants of the compound optical microscope design for specialized purposes. Some of these are physical design differences allowing specialization for certain purposes:

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the sample is illuminated through the objective lens with a narrow set of wavelengths of light. This light interacts with fluorophores in the sample which then emit light of a longer

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extended out to 6 feet to view objects that close. After seeing the compound microscope built by Drebbel exhibited in Rome in 1624, Galileo built his own improved version. In 1625,

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Lee, Joonhee; Crampton, Kevin T.; Tallarida, Nicholas; Apkarian, V. Ara (April 2019). "Visualizing vibrational normal modes of a single molecule with atomically confined light".

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Student microscope – an often low-power microscope with simplified controls and sometimes low-quality optics designed for school use or as a starter instrument for children.

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Additionally, methods such as electron or X-ray microscopy use a vacuum or partial vacuum, which limits their use for live and biological samples (with the exception of an

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obtainable with conventional lenses is about 200 nm. A new type of lens using multiple scattering of light allowed to improve the resolution to below 100 nm.

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Van Putten, E. G.; Akbulut, D.; Bertolotti, J.; Vos, W. L.; Lagendijk, A.; Mosk, A. P. (2011). "Scattering Lens Resolves Sub-100 nm Structures with Visible Light".

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The actual inventor of the compound microscope is unknown although many claims have been made over the years. These include a claim 35 years after they appeared by

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While most techniques focus on increases in lateral resolution there are also some techniques which aim to allow analysis of extremely thin samples. For example,

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to allow viewing of tiny particles whose diameter is below or near the wavelength of visible light (around 500 nanometers); mostly obsolete since the advent of

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Objective turret, revolver, or revolving nose piece is the part that holds the set of objective lenses. It allows the user to switch between objective lenses.

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Courjon, D.; Bulabois, J. (1979). "Real Time Holographic Microscopy Using a Peculiar Holographic Illuminating System and a Rotary Shearing Interferometer".

1284:(NA) of the objective lens. There is therefore a finite limit beyond which it is impossible to resolve separate points in the objective field, known as the 877:

All stages move up and down for focus. With a mechanical stage slides move on two horizontal axes for positioning the specimen to examine specimen details.

1735: 3133: 2681: 3321: 540:, invented the first practical binocular microscope while carrying out one of the earliest and most extensive American microscopic investigations of 2714: 2859: 2097: 1525:

In order to overcome the limitations set by the diffraction limit of visible light other microscopes have been designed which use other waves.

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A simple microscope uses a lens or set of lenses to enlarge an object through angular magnification alone, giving the viewer an erect enlarged

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Cremer, Christoph; Hausmann, Michael; Bradl, Joachim and Rinke, Bernd "Method and devices for measuring distances between object structures",

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The sample can be lit in a variety of ways. Transparent objects can be lit from below and solid objects can be lit with light coming through (

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Optical microscopy is used extensively in microelectronics, nanophysics, biotechnology, pharmaceutic research, mineralogy and microbiology.

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of about 40 to 2 mm, respectively. Objective lenses with higher magnifications normally have a higher numerical aperture and a shorter

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Cremer, Christoph; Hausmann, Michael; Bradl, Joachim and Schneider, Bernhard "Wave field microscope with detection point spread function",

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is a lens designed to focus light from the illumination source onto the sample. The condenser may also include other features, such as a

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methods place the thin sample on a contrast-enhancing surface and thereby allows to directly visualize films as thin as 0.3 nanometers.

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in the resulting image. Some high performance objective lenses may require matched eyepieces to deliver the best optical performance.

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which allows position, distance and angle measurements on "optically isolated" particles (e.g. molecules) well below the theoretical

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Heintzmann, Rainer (1999). Bigio, Irving J.; Schneckenburger, Herbert; Slavik, Jan; Svanberg, Katarina; Viallet, Pierre M. (eds.).

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Despite significant progress in the last decade, techniques for surpassing the diffraction limit remain limited and specialized.

2907: 2648: 1645: 3364: 3359: 3227: 3021: 1372: 588: 2271: 2486: 2451: 2413: 2348: 2159: 2012: 1948: 1712: 1682: 1577:) has been proposed and widely discussed in the literature, but it is not yet competitive with conventional imaging systems. 1991:

J. William Rosenthal, Spectacles and Other Vision Aids: A History and Guide to Collecting, Norman Publishing, 1996, page 391

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William Rosenthal, Spectacles and Other Vision Aids: A History and Guide to Collecting, Norman Publishing, 1996, pp. 391–392

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Atti Della Fondazione Giorgio Ronchi E Contributi Dell'Istituto Nazionale Di Ottica, Volume 30, La Fondazione-1975, page 554

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Digital microscopy with very low light levels to avoid damage to vulnerable biological samples is available using sensitive

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of the object inside the microscope (image 1). That image is then magnified by a second lens or group of lenses (called the

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There are two basic types of optical microscopes: simple microscopes and compound microscopes. A simple microscope uses the

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A. Mark Smith, From Sight to Light: The Passage from Ancient to Modern Optics, University of Chicago Press - 2014, page 387

1226:, or industrial subjects may be a hazard to the objective. Such optics resemble telescopes with close-focus capabilities. 1156: 2523: 1911: 1456:

SPDM (spectral precision distance microscopy), the basic localization microscopy technology is a light optical process of

283:, a widely used variant of epifluorescent illumination that uses a scanning laser to illuminate a sample for fluorescence. 3395: 3326: 2236:

Cassedy JH (1973). "John L. Riddell's Vibrio biceps: Two documents on American microscopy and cholera etiology 1849–59".

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3D dual color super resolution microscopy with Her2 and Her3 in breast cells, standard dyes: Alexa 488, Alexa 568 LIMON

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within such a region all carry different spectral markers (e.g. different colors or other usable differences in the

499:(skopein) meaning "to look at", a name meant to be analogous with "telescope", another word coined by the Linceans. 3306: 3302: 3153: 1553: 286: 112: 1240:

Very small, portable microscopes have found some usage in places where a laboratory microscope would be a burden.

3099: 1114: 604: 584: 312: 308: 289:, used to image fluorescence deeper in scattering media and reduce photobleaching, especially in living samples. 3331: 2048: 1298: 1248: 1728: 73:, slightly different images are used to create a 3-D effect. A camera is typically used to capture the image ( 2195: 1547: 2671: 160:. The use of a single convex lens or groups of lenses are found in simple magnification devices such as the 1535: 328: 108: 928:

and/or filters, to manage the quality and intensity of the illumination. For illumination techniques like

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At very high magnifications with transmitted light, point objects are seen as fuzzy discs surrounded by

3349: 1285: 1142: 1072: 1031: 1011: 973: 870:(rectangular glass plates with typical dimensions of 25×75 mm, on which the specimen is mounted). 274: 69:

on the microscope. In high-power microscopes, both eyepieces typically show the same image, but with a

3316: 3422: 3222: 1481: 650: 316: 268: 116: 3102:, an illustrated collection with more than 3000 photos of scientific microscopes by European makers 2978: 2087: 1977:

Raymond J. Seeger, Men of Physics: Galileo Galilei, His Life and His Works, Elsevier - 2016, page 24

315:, without traditional wavelength-based resolution limits. This microscope primarily realized on the 96:

can be used to increase image contrast by highlighting small details of differing refractive index.

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Laterally modulated excitation microscopy: improvement of resolution by using a diffraction grating

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for specific structures within a cell. In contrast to normal transilluminated light microscopy, in

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graduated to allow measuring distances in the focal plane. The other (and older) type has simple

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light. While larger magnifications are possible no additional details of the object are resolved.

3310: 3292: 3207: 3122: 1529: 1457: 1413: 1383:. In 2005, a microscope capable of detecting a single molecule was described as a teaching tool. 1053: 965: 827: 823: 623: 529: 256: 229:

has two separate light paths allowing direct comparison of two samples via one image in each eye.

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Measuring microscopes are used for precision measurement. There are two basic types. One has a

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image from a sample. Major techniques for generating increased contrast from the sample include

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or water-immersion objectives for greater resolution at high magnification. These are used with

804: 641:, have been used to label specific structures within the cell. More recent developments include 192:

A compound microscope uses a lens close to the object being viewed to collect light (called the

3252: 2973: 2474: 1288:. Assuming that optical aberrations in the whole optical set-up are negligible, the resolution 1180: 921: 896:. Most microscopes, however, have their own adjustable and controllable light source – often a 716: 595:

cells, can be imaged without having to use staining techniques. Just two years later, in 1955,

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with limited magnification, which date at least as far back as the widespread use of lenses in

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SMI (spatially modulated illumination microscopy) is a light optical process of the so-called

909: 561: 505:, another Dutchman, developed a simple 2-lens ocular system in the late 17th century that was 3217: 2391: 2328: 2297: 2002: 1938: 1440:

of the illuminating light, or to extract other structural parameters in the nanometer range.

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in a suitable manner to either increase the optical resolution, to maximize the precision of

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may minimize the risk of damage to the most light-sensitive samples. In this application of

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Objective turret, revolver, or revolving nose piece (to hold multiple objective lenses) (2)

533: 474: 241: 235:, for studying samples from below; useful for cell cultures in liquid or for metallography. 3114: 1214:

In industrial use, binocular microscopes are common. Aside from applications needing true

8: 3297: 3237: 3111:, A collection of 17th through 19th century microscopes, including extensive descriptions 1147: 1095: 619: 393: 303: 280: 232: 2796: 2622: 2571: 1889: 1822: 437:

Compound microscopes first appeared in Europe around 1620 including one demonstrated by

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Fiber optic connector inspection microscope, designed for connector end-face inspection

193: 145: 100: 51: 2860:"High-precision measurements in epifluorescent microscopy – simulation and experiment" 2136:

Daniel J. Boorstin, The Discoverers, Knopf Doubleday Publishing Group - 2011, page 327

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Kumar, Naresh; Weckhuysen, Bert M.; Wain, Andrew J.; Pollard, Andrew J. (April 2019).

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Stimulated emission depletion (STED) microscopy image of actin filaments within a cell

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Four examples of transilumination techniques used to generate contrast in a sample of

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microscopy additional optical components must be precisely aligned in the light path.

21: 3051: 2991: 2987: 2930: 2753: 2740:. Optical Biopsies and Microscopic Techniques III. Vol. 3568. pp. 185–196. 2644: 2630: 2583: 2482: 2344: 2253: 2200: 2155: 2044: 2008: 1944: 1903: 1842: 1834: 1791: 1783: 1708: 1678: 1215: 1200: 1007: 989: 665: 537: 449: 409: 373: 220: 70: 55: 3017: 2942: 2844: 2812: 2645:"Demonstration of a Low-Cost, Single-Molecule Capable, Multimode Optical Microscope" 2595: 2358: 1854: 3267: 3003: 2983: 2922: 2832: 2800: 2741: 2626: 2579: 2575: 2336: 2245: 1893: 1826: 1773: 1624: 1574: 1562: 1376: 1168:

Multiple transmission microscopy for contrast enhancement and aberration reduction.

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and specific contrast-enhanced slides for the visualization of nanometric samples.

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Many techniques are available which modify the light path to generate an improved

557: 3354: 2831:. Optical Biopsies and Microscopic Techniques. Vol. 2926. pp. 201–206. 2378: 2249: 2181: 2148: 2146:

Gould, Stephen Jay (2000). "Chapter 2: The Sharp-Eyed Lynx, Outfoxed by Nature".

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At the lower end of a typical compound optical microscope, there are one or more

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digital cameras. It has been demonstrated that a light source providing pairs of

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technique, placing the specimen on a glass slide and mixing with a salt solution

3247: 2709: 2421: 2340: 1870: 1597: 1474: 1223: 1204: 1159:(where a UV-visible spectrophotometer is integrated with an optical microscope) 1132:

Reflected light, or incident, illumination (for analysis of surface structures)

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Many sources of light can be used. At its simplest, daylight is directed via a

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and a micrometer mechanism for moving the subject relative to the microscope.

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to recognise specific proteins within a sample, and fluorescent proteins like

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The oldest published image known to have been made with a microscope: bees by

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Other microscope variants are designed for different illumination techniques:

184: 3411: 3277: 1907: 1871:"Photon-sparse microscopy: visible light imaging using infrared illumination" 1838: 1787: 1589: 1280:(λ), the refractive materials used to manufacture the objective lens and the 949: 831: 817: 809: 781: 773: 576: 424: 397: 157: 136: 46: 1898: 881:

specimen position adjustments are the reason for having a mechanical stage.

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CLEO:2011 - Laser Applications to Photonic Applications (2011), Paper CThW6

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Using fluorescent samples more techniques are available. Examples include

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Alternatives to optical microscopy which do not use visible light include

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The Lying Stones of Marrakech: Penultimate Reflections in Natural History

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Automation (for automatic scanning of a large sample or image capture)

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Usually a wavelength of 550 nm is assumed, which corresponds to

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of a compound optical microscope is the product of the powers of the

780:. Microscope objectives are characterized by two parameters, namely, 592: 565: 354: 173: 3128: 3077:, Siegfried Weisenburger and Vahid Sandoghdar, arXiv:1412.3255 2014. 517: 144:

microscopes are compound microscopes, while some cheaper commercial

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measurements of fluorescent objects that are small relative to the

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and magnitude of the diffraction patterns are affected by both the

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Modern biological microscopy depends heavily on the development of

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illumination which allows imaging of transparent samples. By using

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is 0.95, and with oil, up to 1.5. In practice the lowest value of

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may be used to determine crystal orientation of metallic objects.

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By Katarina Logg. Chalmers Dept. Applied Physics. 20 January 2006

2329:"Contrast Enhancement by Multi-Pass Phase-Conjugation Microscopy" 1428:(PSF) engineering. These are processes which modify the PSF of a 1230: 541: 3090:

A site about Antique Microscopes, their Accessories, and History

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Since the mid-20th century chemical fluorescent stains, such as

3257: 2908:"Dual color localization microscopy of cellular nanostructures" 2705:"2 Americans and a German Are Awarded Nobel Prize in Chemistry" 1966: 1390: 1027: 893: 709: 365: 2547: 1340: 1277: 905: 705: 277:, designed for analysis of samples that include fluorophores. 165: 2781: 1808: 634: 441:

in London (around 1621) and one exhibited in Rome in 1624.

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and as a result, can achieve much greater magnifications.

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filament, was always visible in the image of the sample.

382: 377: 364:, are also commercially available. These are essentially 3322:

Total internal reflection fluorescence microscopy (TIRF)

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Kenneth, Spring; Keller, H. Ernst; Davidson, Michael W.

2184:, Instituto e Museo di Storia della Scienza (in Italian) 1867: 812:

microscope objective lenses: 100× (left) and 40× (right)

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Van Helden, Albert; Dupre, Sven; Van Gent, Rob (2011).

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spectacle-maker Johannes Zachariassen that his father,

271:, which applies the phase contrast illumination method. 3075:"Light Microscopy: An ongoing contemporary revolution" 2327:

Pégard, Nicolas C.; Fleischer, Jason W. (1 May 2011).

1117:

of different path lengths of light through the sample.

669:

Basic optical transmission microscope elements (1990s)

591:

of light, extremely transparent samples, such as live

1443: 1301: 1252:

The diffraction limit set in stone on a monument for

1075:

illumination, sample contrast comes from rotation of

630:. It is this emitted light which makes up the image. 471:

for the compound microscope Galileo submitted to the

345:

A digital microscope is a microscope equipped with a

25:

Scientist using an optical microscope in a laboratory

3360:

Photo-activated localization microscopy (PALM/STORM)

2001:

Albert Van Helden; Sven Dupré; Rob van Gent (2010).

1937:

Albert Van Helden; Sven Dupré; Rob van Gent (2010).

1670: 2077:

Brian Shmaefsky, Biotechnology 101 - 2006, page 171

749:

Objective turret (revolver or revolving nose piece)

2217:Riddell JL (1854). "On the binocular microscope". 2147: 2145: 2007:. Amsterdam University Press. pp. 32–36, 43. 1704:The Principles and Practice of Electron Microscopy 1646:"Lesson 2 – Page 3, CLASSIFICATION OF MICROSCOPES" 1358: 1328: 376:. The camera is attached directly to a computer's 1487: 912:is often provided on more expensive instruments. 3409: 3088:Antique Microscopes & Scientific Instruments 1861: 1674:The IIT Foundation Series - Physics Class 8, 2/e 1184:A 40x magnification image of cells in a medical 3123:Online tutorial of practical optical microscopy 3018:"German Future Prize for crossing Abbe's Limit" 2696: 2326: 1094:illumination, sample contrast comes from light 385:source or sources adjacent to the camera lens. 2858:Heintzmann, R.; Münch, H.; Cremer, C. (1997). 2670:Ritter, Karl; Rising, Malin (8 October 2014). 2663: 1419: 1347:as the external medium, the highest practical 311:, is a variant of optical microscope based on 65:and may be directly viewed through one or two 3263:Interference reflection microscopy (IRM/RICM) 3154: 2905: 1469:) is being registered. This is possible when 472: 2669: 2512:A discussion of Zeiss measuring microscopes. 1188:taken through an optical microscope using a 211: 2829:Optical Biopsies and Microscopic Techniques 2672:"2 Americans, 1 German win chemistry Nobel" 2479:Encyclopedia of Optical Engineering, Vol. 3 2392:"Long-focus microscope with camera adapter" 2384: 1700: 3161: 3147: 2735: 2235: 2216: 1943:. Amsterdam University Press. p. 24. 1586:environmental scanning electron microscope 995: 799: 610: 575:in physics was awarded to Dutch physicist 2977: 2561: 1897: 1777: 1707:. Cambridge University Press. p. 6. 1329:{\displaystyle d={\frac {\lambda }{2NA}}} 1113:illumination, sample contrast comes from 1056:illumination, sample contrast comes from 726: 265:, similar to the petrographic microscope. 3233:Differential interference contrast (DIC) 1729:"Buying a cheap microscope for home use" 1677:. Pearson Education India. p. 213. 1499: 1447: 1247: 1179: 964: 803: 664: 516: 327: 298:, an adapted light microscope that uses 183: 126: 20: 2472: 2406: 2229: 2210: 1643: 349:allowing observation of a sample via a 3410: 3228:Quantitative phase-contrast microscopy 3168: 2880:from the original on 16 February 2016. 2521: 1696: 1694: 1412:for the development of super-resolved 1373:near field scanning optical microscopy 908:are becoming a more common provision. 547: 179: 3142: 2825: 2702: 2522:Linder, Courtney (22 November 2019). 479:in 1624 (Galileo had called it the " 423:The earliest microscopes were single 323: 3390: 3355:Stimulated emission depletion (STED) 2684:from the original on 11 October 2014 2402:from the original on 3 October 2011. 2308:from the original on 1 November 2008 2100:from the original on 3 February 2017 1480:Many standard fluorescent dyes like 1218:, the use of dual eyepieces reduces 645:, which uses fluorescently labelled 151: 2906:Manuel Gunkel; et al. (2009). 2717:from the original on 9 October 2014 2436: 1691: 1542:Scanning ion-conductance microscopy 1211:on free cells or tissue fragments. 1122: 487:"). Faber coined the name from the 13: 3065: 2541: 2335:. Optica Publishing Group: CThW6. 2302:Olympus Microscopy Resource Center 2272:"How to Use a Compound Microscope" 1444:Localization microscopy SPDMphymod 1026:illumination. A recent technique ( 1024:differential interference contrast 938:differential interference contrast 601:differential interference contrast 319:platforms using all optical tools. 16:Microscope that uses visible light 14: 3439: 3327:Lightsheet microscopy (LSFM/SPIM) 3096:A collection of early microscopes 3081: 3024:from the original on 7 March 2009 2703:Chang, Kenneth (8 October 2014). 2651:from the original on 6 March 2009 1917:from the original on 4 June 2016. 1741:from the original on 5 March 2016 1207:when dealing with tissues, or in 756: 512: 414:Timeline of microscope technology 360:Low-powered digital microscopes, 3389: 3378: 3377: 3276: 3119:, concepts in optical microscopy 3039: 2988:10.1111/j.1365-2818.2010.03436.x 2952:from the original on 3 May 2019. 2477:. In Driggers, Ronald G. (ed.). 1652:from the original on 10 May 2016 1554:Transmission electron microscopy 1103: 1084: 1065: 1046: 943: 772:arrangements are designed to be 701:Stage (to hold the specimen) (6) 690:Focus knobs (to move the stage) 188:Diagram of a compound microscope 113:transmission electron microscopy 3045: 3010: 2956: 2899: 2884: 2851: 2819: 2775: 2760: 2729: 2637: 2602: 2515: 2495: 2466: 2365: 2320: 2289: 2264: 2187: 2168: 2139: 2130: 2121: 2112: 2080: 2071: 2065: 2061: 2054: 2033: 2027: 2021: 1994: 1957: 1520: 1359:Surpassing the resolution limit 1199:Optical microscopy is used for 1175: 887: 874:it may be possible to add one. 579:in 1953 for his development of 313:tip-enhanced Raman spectroscopy 244:, for studying samples of high 3332:Lattice light-sheet microscopy 3243:Second harmonic imaging (SHIM) 3050:. Amsterdam University Press. 2896:priority date 23 December 1996 2580:10.1103/PhysRevLett.106.193905 2088:"Who Invented the Microscope?" 1930: 1921: 1802: 1753: 1721: 1664: 1637: 1488:3D super resolution microscopy 1243: 1135:Fluorescence microscopy, both: 900:, although illumination using 838: 495:(micron) meaning "small", and 131:Diagram of a simple microscope 1: 2414:"Questar Maksutov microscope" 2196:The Lying Stones of Marrakech 1631: 1548:Scanning tunneling microscope 1509:Stimulated emission depletion 1381:stimulated emission depletion 844:stand and had a fixed stage. 822:Some microscopes make use of 660: 309:Tip-enhanced Raman microscope 3048:The Origins of the Telescope 2772:, priority date 10 July 1997 2004:The Origins of the Telescope 1940:The Origins of the Telescope 1536:Scanning electron microscope 960: 915: 536:, Professor of Chemistry at 418: 109:scanning electron microscopy 7: 2481:. CRC Press. p. 2409. 2444:"FTA long-focus microscope" 2175:"Il microscopio di Galileo" 1607: 1420:Structured illumination SMI 10: 3444: 3125:at University of Cambridge 2631:10.1088/0150-536X/10/3/004 2341:10.1364/CLEO_SI.2011.CThW6 2250:10.1093/jhmas/xxviii.2.101 2193:Gould, Stephen Jay (2000) 2154:. New York, N.Y: Harmony. 1671:Trisha Knowledge Systems. 1143:Epifluorescence microscopy 999: 974:Office of Field Operations 815: 760: 730: 678:Eyepiece (ocular lens) (1) 607:-based imaging technique. 407: 403: 338: 275:Epifluorescence microscope 61:The object is placed on a 3373: 3340: 3285: 3274: 3198: 3176: 2805:10.1007/s00340-008-3152-x 1831:10.1038/s41586-019-1059-9 1779:10.1038/s41596-019-0132-z 1477:of different particles). 1203:, the field being termed 1079:light through the sample. 599:published the theory for 473: 372:and generally do not use 317:scanning-probe microscope 269:Phase-contrast microscope 212:Other microscope variants 117:scanning probe microscopy 2473:Ollsson, Gustaf (2003). 2039:Shmaefsky, Brian (2006) 1398:Nobel Prize in Chemistry 1263:rings. These are called 1165:Near-Infrared microscopy 860: 847: 824:oil-immersion objectives 122: 33:, also referred to as a 3293:Fluorescence microscopy 3253:Structured illumination 3208:Bright-field microscopy 3094:Antique Microscopes.com 2550:Physical Review Letters 2377:24 January 2011 at the 2298:"Microscope objectives" 1899:10.1364/OPTICA.2.001049 1530:Atomic force microscope 1495: 1458:fluorescence microscopy 1414:fluorescence microscopy 1396:On 8 October 2014, the 1060:of light in the sample. 996:Illumination techniques 828:index-matching material 800:Oil immersion objective 657:making it fluorescent. 624:fluorescence microscopy 611:Fluorescence microscopy 530:Antonie van Leeuwenhoek 257:Petrographic microscope 196:lens), which focuses a 3365:Near-field (NSOM/SNOM) 3303:Multiphoton microscopy 3134:Cell Centered Database 3100:Historical microscopes 2927:10.1002/biot.200900005 1559:Ultraviolet microscope 1505: 1453: 1330: 1256: 1193: 1162:Ultraviolet microscopy 992: 813: 727:Eyepiece (ocular lens) 670: 653:which a live cell can 526: 336: 189: 132: 94:Phase-contrast imaging 88:) the objective lens. 26: 3218:Dark-field microscopy 3116:Molecular Expressions 2966:Journal of Microscopy 2915:Biotechnology Journal 2893:U.S. patent 6,424,421 2769:U.S. patent 7,342,717 2448:firsttenangstroms.com 2372:O1 Optical Microscopy 2096:. 14 September 2013. 2043:. Greenwood. p. 171. 1734:. Oxford University. 1503: 1451: 1426:point spread function 1331: 1251: 1183: 1073:Cross-polarized light 1032:cross-polarized light 1012:cross-polarized light 986:international airport 968: 807: 693:Coarse adjustment (4) 668: 520: 434:in the 13th century. 331: 287:Two-photon microscope 263:Polarizing microscope 227:Comparison microscope 187: 130: 24: 3286:Fluorescence methods 3109:The Golub Collection 2510:. 1906. p. 716. 2180:9 April 2008 at the 1701:Ian M. Watt (1997). 1299: 1292:, can be stated as: 948:The actual power or 722:Mechanical stage (9) 603:microscopy, another 534:John Leonard Riddell 475:Accademia dei Lincei 368:with a high-powered 304:electron microscopes 242:Traveling microscope 3317:Image deconvolution 3298:Confocal microscopy 3238:Dispersion staining 3213:Köhler illumination 2797:2008ApPhB..93....1L 2623:1979JOpt...10..125C 2572:2011PhRvL.106s3905V 2454:on 26 February 2012 1890:2015Optic...2.1049A 1823:2019Natur.568...78L 1620:Köhler illumination 1462:limit of resolution 1148:Confocal microscopy 976:agent checking the 910:Köhler illumination 696:Fine adjustment (5) 562:Köhler illumination 548:Lighting techniques 281:Confocal microscope 233:Inverted microscope 180:Compound microscope 146:digital microscopes 41:that commonly uses 3428:Optical microscopy 3189:Optical microscopy 3170:Optical microscopy 1615:Digital microscope 1506: 1454: 1326: 1282:numerical aperture 1257: 1194: 993: 814: 786:numerical aperture 763:Objective (optics) 671: 643:immunofluorescence 527: 523:Francesco Stelluti 503:Christiaan Huygens 428:magnifying glasses 341:Digital microscope 337: 324:Digital microscope 246:optical resolution 190: 133: 31:optical microscope 27: 3405: 3404: 3350:Diffraction limit 2837:10.1117/12.260797 2785:Applied Physics B 2746:10.1117/12.336833 2611:Journal of Optics 2528:Popular Mechanics 2488:978-0-824-74252-2 2350:978-1-55752-910-7 2199:. Harmony Books. 2161:978-0-224-05044-9 2041:Biotechnology 101 2014:978-90-6984-615-6 1950:978-90-6984-615-6 1714:978-0-521-43591-8 1684:978-81-317-6147-2 1648:. msnucleus.org. 1324: 1286:diffraction limit 1201:medical diagnosis 1157:Microspectroscopy 1042:. 1.559 μm/pixel. 990:stereo microscope 538:Tulane University 450:Zacharias Janssen 410:History of optics 394:entangled photons 374:transillumination 221:Stereo microscope 176:and microscopes. 152:Simple microscope 71:stereo microscope 3435: 3423:Dutch inventions 3393: 3392: 3381: 3380: 3343:limit techniques 3280: 3201:contrast methods 3199:Illumination and 3163: 3156: 3149: 3140: 3139: 3105: 3061: 3034: 3033: 3031: 3029: 3014: 3008: 3007: 2981: 2960: 2954: 2953: 2951: 2912: 2903: 2897: 2895: 2888: 2882: 2881: 2879: 2864: 2855: 2849: 2848: 2823: 2817: 2816: 2779: 2773: 2771: 2764: 2758: 2757: 2733: 2727: 2726: 2724: 2722: 2700: 2694: 2693: 2691: 2689: 2667: 2661: 2660: 2658: 2656: 2641: 2635: 2634: 2606: 2600: 2599: 2565: 2545: 2539: 2538: 2536: 2534: 2519: 2513: 2511: 2499: 2493: 2492: 2470: 2464: 2463: 2461: 2459: 2450:. 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439:Cornelis Drebbel 300:light scattering 162:magnifying glass 45:and a system of 35:light microscope 3443: 3442: 3438: 3437: 3436: 3434: 3433: 3432: 3408: 3407: 3406: 3401: 3369: 3342: 3341:Sub-diffraction 3336: 3281: 3272: 3200: 3194: 3172: 3167: 3103: 3084: 3068: 3066:Further reading 3058: 3042: 3037: 3027: 3025: 3016: 3015: 3011: 2979:10.1.1.665.3604 2961: 2957: 2949: 2910: 2904: 2900: 2891: 2889: 2885: 2877: 2862: 2856: 2852: 2824: 2820: 2780: 2776: 2767: 2765: 2761: 2734: 2730: 2720: 2718: 2701: 2697: 2687: 2685: 2668: 2664: 2654: 2652: 2643: 2642: 2638: 2607: 2603: 2546: 2542: 2532: 2530: 2520: 2516: 2501: 2500: 2496: 2489: 2471: 2467: 2457: 2455: 2442: 2441: 2437: 2427: 2425: 2424:on 15 June 2011 2412: 2411: 2407: 2390: 2389: 2385: 2379:Wayback Machine 2370: 2366: 2351: 2325: 2321: 2311: 2309: 2294: 2290: 2280: 2278: 2270: 2269: 2265: 2234: 2230: 2219:Q J Microsc Sci 2215: 2211: 2192: 2188: 2182:Wayback Machine 2173: 2169: 2162: 2144: 2140: 2135: 2131: 2126: 2122: 2117: 2113: 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