History and Types of Microscopes

report and Types of MicroscopesWhat is a microscope?There is so many little objects that human look pratt be able to see. The microscope is a tool to see dainty objects consisting of lens or combination of lenses1. Due to their soaringly- alter lenses, we go off observe racy-quality word pictures and these age this material bodys stooge be transferred to computers. Todays microscopes argon so advanced that they give the bounce supply up objects which ar sized of the millionth part of a meter called micron2.The kindling of searching minuscule objects with microscopes is called microscopy. Microscopic means that impossible to see, without a aid of a microscope, with a naked tenderness3.History of MicroscopeAfter the glass is start institute in the counterbalance century, Romans was trying to make objects to be seen bigger. The kickoff and simple forms were called flea glasses and they were able to show 6 times bigger4.The microscope is true in Netherlands at th e 1590s further its inventor is not easy to identify. any(prenominal) proofs atomic number 18 leading to Cornelis Drebbel5. But others insist that Zacharias Jansen and his father Hans were working with lenses, they have roughly lenses and put them into a tube and invented the microscope. Few others believed that Galileo Galilei was the first sentinel of microscope6.First microscopes were not favorable enough to mathematical function at interrogationes because it crowd out only enlarge by 9 times bigger7.First, the real microscope was employ by Anton van Leeuwenhoek in the late 17th century which was made by pipes, simple lens, plate and screw( intiree1). trope 1Unlike the others, his microscope could show objects whizz-millionth of a meter bigger of its sizes(270x). Others best achievement was 50x magnification. With this microscope, he saw and identified bacteria, erythrocyte, and sperm cells. He published their drawings on philosophical Transactions of the Royal Societ y of London at 1674.These drawings were forgotten until at that place were huge developments in science8.In 1665 van Leeuwenhoeks work was a demand to Robert Hooke and he wrote Micrographia. It is the first book that provides microscopic pictures of bird louses, industrial plants etc. 9 ( visualize 2).Figure 2-Drawing of an insect by Robert Hooke10After 200 years from Robert Hooke, German engineer called Carl Zeiss improved lenses of the microscope and he established a company named Zeiss. After that, he hired Ernst Abbe to the company. Abbe improved the microscopes and lenses11.Types of microscopesStereoscopeDissection microscope is used with visible brightness. It is used to see dissection better.It has 3-dimensional renders and it has crushed magnification.Figure 3 earthworm captured by StereoscopeConfocal MicroscopeConfocal laser s lowlifening microscopy (CLSM) plays the nearly momentous role on imaging tiny try ons in three-dimensional form. CLSM whole kit and ca boodle like an optic microscope with some differences. It uses monochromatic laser exculpated kinda of visible out of work 12.CLSM has widely used from cell biology, genetics, microbiology and development biology to quantum optics, nanocrystal imaging and spectroscopy13.History of Confocal MicroscopeEarly in 1940, Hans Goldmann from Switzerland invented a slit lamp to make documentation of eye examinations. Some researchers believe it might be first confocal optic system 14.Marvin Minsky invented first confocal scan microscope in 1955 and in 1957 got its patent.Figure 4 Marvin Minskys patent application that shows the principle of CLSM 15.By moving the stage, blaze mind in focal plane could be scanned 16.In 1969 M. David egger and Paul Davidovits described the first CLSM in two pages and published. Only one illumination spot generated with this point scanner. It was used for the imaging of the nerve interweave 17, 18.In 1983 confocal microscope was first used and controlled by a computer afterwards the publication of first work by I. J. Cox and C. Sheppard from Oxford University. ascendantd on Oxford groups designs, first CLSM was offered from 1982 19.At the Laboratory of Molecular Biology in Cambridge, William Bradshaw Amos and washstand Graham White and colleagues invented the first confocal shine scanning microscope in the warmheartedness of 1980s.This time the illumination spot was moving tho not the stage. This proficiency allowed faster work out acquisition, quartette images per plump for 20.Working Principle of Confocal MicroscopeFor hireting higher intensities a laser is used. The laser light think overs from the dichroic mirror. After that it hits mirrors on motors and a penetrate the strain lasers trip up scanned by these mirrors. And emitted light passes through the dichroic mirror and gets focused onto pinhole. Finally, the detector measures that light. As it appears the complete image of the sample cannot be observed just on e point can be observed. The photomultiplier detector is connected to a computer and one pixel at a time it builds an image 21.Figure 5 Principal Light Pathways in Confocal Microscopy 22.What is the advantage of using a confocal microscope?By scanning lots of smooth parts of a sample, it is easy to build a very unspoilt three-dimensional image. Confocal microscope has better resolvent horizontally and vertically. The best dissolvent can be obtained at 0.2 microns for horizontal and 0.5 microns for vertical 23. fashion modelsThere be some examples of imaging with the confocal microscope. Figure 6 Nematode. Miami University in Oxford, Ohio 24.Figure 7 Example image of confocal microscope 25.Scanning Electron Microscope (SEM)SEM is an negatron microscope that uses the focused beam of electrons to images of the sample. Electrons move with atoms in the sample and gives information about external morphology (texture), chemic composition, and crystalline structure and orientation of materials making up the sample 26.A beam of electrons uses raster scan pattern which is a rectangular pattern of an image and reconstructive memory in the screen. Most computers use bitmap image systems to store the image 27.The image is created by matching the position with the perceived signal. SEM can get better than 1 nm resolution. Standard SEM microscopes argon generally qualified for dry and conductive surfaces in high vacuum. Also, there are specialize machines that work chthonian alternateable conditions from low temperature to high temperature and in low vacuum. There is environmental SEM for wet conditions.McMullan presented the history of SEM 28. Manfred von Ardenne invented SEM in 1937. In the primeval 1960s, Cambridge groups marketed as Stereoscan in 196528, 29.After interaction of high energized beam of electrons and outermost orbit electrons of samples atoms Auger electrons which have low electrons pull up stakes be formed. These electrons exculpate inform ation about sample surface.After interactions, there go forth be electron beams which have lower energy, move to the surface of the sample and will acquire there.These electrons called as secondary electrons. For imaging for SEM, mostly secondary electrons are world used. Change of secondary electrons numbers depends on the topography of surface and angle of the point where the beam hits the surface 30.Figure 7 Blood image by SEM 31.Transmission Electron MicroscopeHigh energized electrons pass through the very handsome sample. After interaction of electrons, images are enlarged and focused on fluorescence screen, photographic film layer or CCD camera 32.In 1930 Max hillock and Ernst Ruska invented TEM 33. It allows us to see smaller objects than the optical microscope.TEM is used in malignant neoplastic disease research, virology, materials science, nanotechnology, and semiconductor.TEMs contrast depends on absorption of electrons, thickness, and composition of the sample. Compl ex wave interactions at higher magnifications modulate the intensity of the image with analysis of an expert for the image. The resolution determine is up to 0.2 nm for TEM.Compared to SEM, TEM has troublesome work to get the sample ready and the user must have a very good background about it 34.Figure 8 Example of TEM of a plant cell 35.Compound Light MicroscopesCompound microscopes are 2-dimensional light microscopes and they are most used microscopes. Even though it has low resolution it has high magnification.Figure 9-meiosis seen by compound microscope36.Figure 10-Microscope view of plant cells37. part of Optical MicroscopeFigure 10 Parts of a microscope38Eyepiece lense The lens that allows us to see through.Tubes It helps eyepiece to connect to lenses.Arm Holds the tube.Base Supports the microscope at the bottom.Illuminator Light source or a mirror that helps us to see a sample from the tube. If it is a mirror it can reflect outer light to use.Stage This platform is used to put samples and it has clips to save the sample from moving.Revolving Nosepiece or Turret This part is for holding lenses together and it can rotate to switch surrounded by lenses.Objective Lenses These lenses are most commonly can be put three or four lenses on the microscope. They have 4,10,40 or nose candy times bigger magnification. They are color coded and should build to DIN standards.Rack Stop It is used to harbor the target lens from breaking39.DIN StandardsThe real image is formed clxmm out from the prey lens.Parfocal distance should be 45 mm.Eyepiece lens should be 170mm40.Working Principle of Optical MicroscopeFigure 11 41As shown in Figure 9 light starts its journey from illuminator and with a mirror it reaches to sample. thus it goes to prism through verifiable lenses. It reflects from the prism and comes to eye in the tube. When light passes through the objective lens makes the image of sample bigger and focuses 160 mm inside the tube and and so ocular lense s magnifies the image of sample 25cm away from the eye. This image is a virtual image of the sample (Figure 10). ordinary microscopes have four different objective lenses. Scanning (5x), low office staff(10x), strong point power (20x) and high power lenses (40x). We can easily calculate the magnifying of the microscope with multiplying objective lens and ocular lens. For example, after image magnified by objective lenses 40 times of original image of the sample, will magnify second time 20 times bigger by ocular lenses. So, our eye can see 4020=800 times bigger image of an original image of the sample.Figure 12 42Differences Between Electron and Light MicroscopeLight microscopes techniques are simple merely for electron microscope high-level technical skill needed. facility time of the sample is few minutes to few hours for light microscopes but some(prenominal) days for electron microscopes.Live or deathlike samples can be seen in light microscopes but for electron microscope s only dead and dried samples can be seen.Light microscopes have low resolution than electron microscope and the resolution margin for the light microscope is 200 nm but for SEM 1nm and for TEM 0.2 nm.Light rays are used to illuminate for light microscope but for electron microscope electrons are being used.Lenses are made of glass for light microscope but for electron microscope all lenses are electromagnets.Magnification of light microscope is 500x to 1500x but for EM 160,000x and photographic magnification is 1000,000x or more.Light microscopes are cheap but electron microscopes are expensive 43.Calculation of ResolutionIf we want to get good details of very small objects like cells, we need to gain the resolution. It can be described as to see different between two small and very near objects. It can be change of the wavelength of light and power of lenses. Mathematical formula of separating two different small objects which have the smallest distance (dmin)Dmin = 1.22 x wave length / N.A. objective + N.A. condenserDifferent then the theoretical power, in employ samples quality affects its resolving power44.Definition of numeral Aperture(N.A.) is a value of objectives defined by Abbe.Numerical Aperture (NA)=n-sin() or n-sin()Figure 13 Numerical ApertureAs shown in Figure 11 light waves go through a sample to the objective lens. But when it comes to practice it is nearly impossible to get the value of aperture above 0.95 with dry objective lenses. When the light cones get the bigger degree of starts to increase from 7 to 60 and N.A. increases from 0.12 to 0.87. In todays world, it is possible to use alternative media to make images in water system (refractive index = 1.33), glycerin (refractive index = 1.47), and immersion oil (refractive index = 1.51) by the objective lens. We can clearly see Figure 12 and remand 1 highly corrected objectives have bigger N.A.Figure 14 prorogue 1 Numerical Aperture versus Optical Correction45There is a repair of re solution in optical microscopes as shown belowLet N.A. be 1.4 and resolution is different for lights wavelength.A minimum distance of two points of the image is 0.61 /N.A.As we know visible light wavelength is between 400-700 nm.There will be no resolution between two objects if distance is 1/3 .If we conduct green light = 500nm and r=0.61 x 500nm / 1.4 =218 nm.If we choose blue light = 400nm and r=0.61 x 400nm / 1.4 =174 nm.If we choose green light = 700nm and r=0.61 x 700nm / 1.4 =305 nm46.Diffraction condition of Electron MicroscopeElectron microscope has diffraction limit and it is 1nm for SEM, 0.3nm for TEM. This limit occurs because of wave nature of electrons. Electrons has a phenomenon called wave-particle duality. Particle of matter (incident electron) can be explained as wave. We can assimilate to sound or water waves.Louis de de Broglie says that the wavelength of a particle can be calculated as sideline formula=h/p wavelength of a particleh Plancks eonian (62610-3 4)p momentum of a particleMomentum is the product of muckle and the f number of a particle and equation can be create verbally as= h / mvAccelerating voltage determines the velocity of the electrons we can use following formulaeV = mv2/2We can calculate the velocity of electrons byDue to these formulae, we can show the wavelength of propagating electrons at a prone accelerating voltageSince the mass of an electron is 9.1 x 10-31 kg and e = 1.6 x 10-19So, the wavelength of electrons is 3.88pm when the microscope is operating at 100 keV, 2.74 pm at 200 keV, and 2.24 pm at 300 keV.We know electrons in an electron microscope reach %70 of speed of the light wit accelerating voltage of 200 keV, there are effects which are significant length contraction, time dilation, and an increase in mass. By these changesc speed of the light (299 792 458 mps)So, wavelength of an electron at 100 keV, 200 keV, 300 keV in electron microscopes is 3.70 pm ,2.51 pm, and 1.96 pm, respectively 47. some ot her reason for limitation for TEM is, sample transparency has to be proper for electron transparency. To be more precise its thickness has to be 100nm or less.Electrons can be deflected in magnetic fields by the Lorentz force. This problem may make crystal structure determination virtually impossible 48, 49.Diffraction Limit of Optical MicroscopeThere is a limit for imaging with an optical microscope called Abbe diffraction limit. This limit is /2( is imaging radiations free-space wavelength) 50. Modern works show us that this limit can be passed and can make optical microscopes lenses to have a high resolution51.But with diffraction limit even though the lens is corrected there will be blur image of the point. This called Airy dish or diffraction. British mathematician Lord George Biddel Airy has found it. We can see its cross section and appearance below (Figure 13).Figure 15Diameter of the disk isBdiff =2.44 (f/)52With f/ limitation can be controlled and wavelength of the ligh t. The muckimum resolving power of the lens is determined by this limitation. If we want to calculate diffraction limit we can use following formulaIf we reach the limit lens will become unable to resolve greater frequencies. In theory, if the contrast is %0 the diffraction limit will appear to be as shown in Table 2 at different f/s for 0.520 m light as known as green light.Table 253Different ship canal to Break Resolution Limit of Optical MicroscopeThere are several ways to break resolution limit of optical microscope. To do that researchers change lenses or different parts of microscopes. Here are some examplesBy employing stimulated electric discharge to inhibit the fluorescence process in the outer regions of the discomfort point-spread function54.By using laterally structured illumination in a wide-field, non-confocal microscope(This method claims that spatially structured excitation light illuminates the sample) 55.By amend the lenses with ZrO2.Synthesis of ZrO2 Nanoparti clesZirconium(IV) isopropoxide2-propanol complex (5.6 g) and anhydrous benzyl alcohol (55mL) were charged into a 100 mL Teflon-lined autoclave. This Teflon-lined autoclave was sealed and placed into an oven at 240 C for 4 days and then cooled to obtain a white mucky suspension. 56.Figure 1657.Figure 16 is a schematic of hSIL integrated with an Olympus optical microscope for super-resolution imaging of the underlying nanopattern. The hSIL collects near-field information on the nanopattern and forms a virtual image that can be captured by the objective lens57.Figure 17 -Super-resolution optical imaging through hSIL on 45 nm gaps. SEM images of the break short with biweekly structures of 50 nm gaps (a) and the gold-coated chip with 45 nm gaps (b). (c, d) Optical images of the chip with 50 nm gaps under white and filtered blue light (max 470 nm) without SILs. (e1, e2) Optical images of the chip with hSIL of h/d = 0.8 (d = 11.5 m). (f1, f2) Optical images of the gold-coated chips thr ough SIL of h/d = 0.78 (d = 10.5 m) and (g1, g2) with hSIL of higher h/d = 0.84 (d = 11.3 m). Optical images of e1g1 and e2g2 were taken under white light and filtered blue light, respectively. The corresponding image magnification factors of e2, f2, and g2 are 3.1, 2.9, and 3.6. The scale bars for e1g2 are the same as that of c58.References1.http//www.life.umd.edu/cbmg/ efficiency/wolniak/wolniakmicro.html2.http//www.kurallarinelerdir.com/2016/04/mikroskop-nedir-mikroskobun-tarihi.html3.https//en.wikipedia.org/wiki/Microscope4.http//www.history-of-the-microscope.org/history-of-the-microscope-who-invented-the-microscope.php5.Albert Van Helden, S.D., Rob Van Gent, Huib Zuidervaart, The Origins of the Telescope. 2010.6.Jay, S., Chapter 2 The Sharp-Eyed Lynx, Outfoxed by Nature. The Lying Stones of Marrakech penult Reflections in Natural History, 2000.7.http//kanbilim.com/?p=1938.http//www.history-of-the-microscope.org/history-of-the-microscope-who-invented-the-microscope.php9.https// en.wikipedia.org/wiki/Micrographia10.http//www.bl.uk/learning/timeline/large107702.html11.http//www.zeiss.com/corporate/int/history/founders.html12.Littlejohn, G.R., et al., Perfluorodecalin enhances in vivo confocal microscopy resolution of Arabidopsis thaliana mesophyll. New Phytologist, 2010. 186(4) p. 1018-1025.13.Hoffman, A., et al., Confocal laser endomicroscopy technical status and reliable indications. Endoscopy, 2006. 38(12) p. 1275-1283.14.Goldmann, H., Spaltlampenphotographie und photometric. Ophthalmologica, 1939. 98(5-6) p. 257-270.15.Minsky, M., Microscopy Apparatus. US Patent 1961. 3.013.467.16.Minsky, M., Memoir on inventing the confocal scanning microscope. Scanning, 1988. 10(4) p. 128-138.17.Davidovits, P. and M.D. Egger, Scanning laser Microscope. Nature, 1969. 223(5208) p. 831-831.18.Davidovits, P. and M.D. Egger, Scanning Laser Microscope for Biological Investigations. Applied Optics, 1971. 10(7) p. 1615-1619.19.Cox, I.J. and C.J.R. Sheppard, Scanning optical microscope incorporating a digital framestore and microcomputer. Applied Optics, 1983. 22(10) p. 1474-1478.20.White, J.G., W.B. Amos, and M. Fordham, An evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy. The daybook of Cell Biology, 1987. 105(1) p. 41-48.21.http//www.physics.emory.edu/faculty/weeks//confocal/22.https//www.microscopyu.com/techniques/confocal/introductory-confocal-concepts23.Prasad, V., D. Semwogerere, and R.W. Eric, Confocal microscopy of colloids. Journal of Physics Condensed Matter, 2007. 19(11) p. 113102.24.http//www.cas.miamioh.edu/mbi-ws/microscopes/confocal.html25.http//depts.washington.edu/keck/intro.htm26.http//serc.carleton.edu/research_education/geochemsheets/techniques/SEM.html27.Leblanc, M., Etude sur la transmission lectrique des impressions lumineuses. La Lumire lectrique, 1880.28.McMullan, D. An improved scanning electron microscope for opaque specimens. Proceedings of the IEE Part II Power E ngineering, 1953. 100, 245-256.29.von Ardenne, M., rabbit Elektronen-Rastermikroskop. Zeitschrift fr Physik, 1938. 109(9) p. 553-572.30.Smith, K.C.A. and C.W. Oatley, The scanning electron microscope and its fields of application. British Journal of Applied Physics, 1955. 6(11) p. 391.31.http//metassoc.com/services/scanning-electron-microscopy/sem-eds-application-examples/32.Crewe, A.V., J. Wall, and J. Langmore, Visibility of Single Atoms. Science, 1970. 168(3937) p. 1338-1340.33.http//www.nobelprize.org/nobel_prizes/physics/laureates/1986/perspectives.html34.Meyer, J.C., et al., Imaging and dynamics of light atoms and molecules on graphene. Nature, 2008. 454(7202) p. 319-322.35.http//www.vcbio.science.ru.nl/en/image-gallery/electron/36.http//www.cas.miamioh.edu/mbi-ws/microscopes/types.html37.http//ichef.bbci.co.uk/images/ic/640xn/p023r74v.jpg38.http//www.microscope-microscope.org/ staple fibre/microscope-parts.htm39.http//www.microscope-microscope.org/basic/microscope-parts.htm4 0.http//www.din.de/en41.DEVEC, D.D.E., MKROSKOP ETLER ALIMA PRENSPLER. Dicle Universitesi.42.https//www.cs.mcgill.ca/rwest/wikispeedia/wpcd/wp/o/Optical_microscope.htm43.http//www.biologyexams4u.com/2012/10/difference-between-light-microscope-and.html44.http//www.life.umd.edu/cbmg/faculty/wolniak/wolniakmicro.html45.https//www.microscopyu.com/microscopy-basics/numerical-aperture46.http//www.math.ubc.ca/cass/courses/m309-03a/m309-projects/cannon/resolving2.html47.Bendersky, L.A. and F.W. Gayle, Electron diffraction using transmission electron microscopy. Journal of research of the National Institute of Standards and Technology, 2001. 106(6) p. 997.48.Thomson, G.P. and A. Reid, Diffraction of cathode rays by a thin film. Nature, 1927. 119 p. 890.49.Thomas, G. and M.J. Goringe, Transmission electron microscopy of materials. 1979.50.Abbe, E., Arch. Mikrosk. Anat. 1873.51.Hecht, L.N.a.B., Principles of Nano-Optics. Cambridge U Press, 2006.52.Riedl, M.J., Optical Design Fundamentals for I nfrared Systems, Second Edition. SPIE Press, Bellingham, WA 2001.53.http//www.edmundoptics.com/resources/application-notes/imaging/diffraction-limit/54.Hell, S.W. and J. Wichmann, Breaking the diffraction resolution limit by stimulated emission stimulated-emission-depletion fluorescence microscopy. Optics Letters, 1994. 19(11) p. 780-782.55.Gustafsson, M.G.L., Surpassi