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Advances in Microscopy and Its Applications with Special Reference to Fluorescence Microscope: An Overview

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Biomedical Visualisation

Part of the book series: Advances in Experimental Medicine and Biology ((AEMB,volume 1406))

Abstract

The microscope has revolutionized the understanding of an organism’s structural details and cellular functions. With the invention of highly evolved microscopes, the diagnosis and treatment of diseases has gained momentum. Technology has immensely helped demonstrate cellular events like phagocytosis, cell movement, cell division, etc. with enhanced temporal and spatial resolution. One of these advanced inventions is the fluorescent microscope which has enabled scanning through various physiological activities of the cell. A fluorescence microscope uses the property of fluorescence to create an image. In addition to visualizing the structural details of the cells, a fluorescence microscope also aids in witnessing cellular activities. With an immunofluorescence microscope, cellular antigens can be localized. This chapter highlights the basics of microscopy, types of microscopes, principles, and types of fluorescence microscopes, and recent advances in microscopy and its application.

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References

  • Abbe E (1874) A contribution to the theory of the microscope and the nature of microscopic vision. Proc Bristol Nat Soc 1:200–261

    Google Scholar 

  • Abramowitz M, Davidson MW (2007) Introduction to microscopy. Molecular Expressions. Retrieved June 30, 2022

    Google Scholar 

  • Ambrose EJ (1956) A surface contact microscope for the study of cell movements. Nature 178:1194

    Article  CAS  PubMed  Google Scholar 

  • Atti Della, Fondazione Giorgio, Ronchi E (1975) Contributi Dell’Istituto Nazionale Di Ottica, vol 30, La Fondazione, p 554

    Google Scholar 

  • Axelrod D (2001) Total internal reflection fluorescence microscopy in cell biology. Traffic 2:764–774

    Article  CAS  PubMed  Google Scholar 

  • Bell AG (1880) On the production and reproduction of sound by light. Am J Sci s3-20(118):305–324

    Article  Google Scholar 

  • Betzig E, Patterson GH, Sougrat R, Lindwasser OW, Olenych S, Bonifacino JS et al (2006) Imaging intracellular fluorescent proteins at nanometer resolution. Science 313:1642–1645

    Article  CAS  PubMed  Google Scholar 

  • Brakenhoff GJ, Voort HVD, Spronsen EV, Nanninga N (1986) Three-dimensional imaging by confocal scanning fluorescence microscopy a. Ann N Y Acad Sci 483(1):405–415

    Article  CAS  PubMed  Google Scholar 

  • Bruni M, Cimini F, Costa U (1998) An alternative method of detecting dispersion staining colors for the determination of asbestos fibers in bulk materials. Med Lav 89(3):254–264

    CAS  PubMed  Google Scholar 

  • Chandler DE, Roberson RW (2009) Bioimaging: current concepts in light and electron microscopy. Jones & Bartlett Publishers

    Google Scholar 

  • Coumans FAW, Van der Pol E, Terstappen LWMM (2012) Flat-top illumination profile in an epi-fluorescence microscope by dual micro lens arrays. Cytometry A 81(4):324–331

    Article  PubMed  Google Scholar 

  • Dani A, Huang B (2010) New resolving power for light microscopy: applications to neurobiology. Curr Opin Neurobiol 20(5):648–652

    Article  CAS  PubMed  Google Scholar 

  • Denk W, Strickler JH, Webb WW (1990) Two-photon laser scanning fluorescence microscopy. Science (New York, NY) 248(4951):73–76

    Article  CAS  Google Scholar 

  • Denk W, Delaney KR, Gelperin A, Kleinfeld D, Strowbridge BW, Tank DW, Yuste R (1994) Anatomical and functional imaging of neurons using 2-photon laser scanning microscopy. J Neurosci Methods 54(2):151–162

    Article  CAS  PubMed  Google Scholar 

  • Dinelli F, Albonetti C, Kolosov OV (2011) Ultrasonic force microscopy: detection and imaging of ultra-thin molecular domains. Ultramicroscopy 111(4):267–272

    Article  CAS  PubMed  Google Scholar 

  • Duarte FJ (2016) Tunable laser microscopy. In: Duarte FJ (ed) Tunable laser applications, 3rd edn. CRC Press, Boca Raton, FL, pp 315–328

    Chapter  Google Scholar 

  • Gill H (2010) Evaluating the efficacy of tryptophan fluorescence and absorbance as a selection tool for identifying protein crystals. Acta Crystallogr F66(Pt 3):364–372

    Google Scholar 

  • Gustafsson MG, Shao L, Carlton PM, Wang CR, Golubovskaya IN, Cande WZ et al (2008) Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination. Biophys J 94(12):4957–4970

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hama H, Kurokawa H, Kawano H, Ando R, Shimogori T, Noda H, Fukami K, Sakaue-Sawano A, Miyawaki A (2011) Scale: a chemical approach for fluorescence imaging and reconstruction of transparent mouse brain. Nat Neurosci 14:1481–1488

    Article  CAS  PubMed  Google Scholar 

  • Heimann PA, Urstadt R (1990) Deep ultraviolet microscope. Appl Opt 29(4):495–501

    Article  CAS  PubMed  Google Scholar 

  • Hell SW (2009) Microscopy and its focal switch. Nat Methods 6:24–32

    Article  CAS  PubMed  Google Scholar 

  • Helmchen F, Denk W (2005) Deep tissue two-photon microscopy. Nat Methods 2:932–940

    Article  CAS  PubMed  Google Scholar 

  • Huang B (2010) Super resolution fluorescence microscopy. Annu Rev Biochem 78:993–1016

    Article  Google Scholar 

  • Im K, Mareninov S, Diaz M, Yong WH (2019) An Introduction to Performing Immunofluorescence Staining. Methods Mol Biol (Clifton, N.J.) 1897:299–311

    Article  CAS  Google Scholar 

  • Inoué S (2006) Foundations of confocal scanned imaging in light microscopy. In: Pawley J (ed) Handbook of biological confocal microscopy. Springer, Boston, MA

    Google Scholar 

  • Jonkman JEN, Swoger J, Kress H, Rohrbach A, Stelzer EHK (2003) Resolution in optical microscopy. Methods Enzymol 360:416–446

    Article  CAS  PubMed  Google Scholar 

  • Kosasih FU, Ducati C (2018) Characterizing degradation of perovskite solar cells through in-situ and operando electron microscopy. Nano Energy 47:243–256

    Article  CAS  Google Scholar 

  • Lakowicz JR (2006) Principles of fluorescence spectroscopy (3rd ed.). Springer, p 954. ISBN 978-0-387-31278-1

    Google Scholar 

  • Lane N (2015) The unseen world: reflections on Leeuwenhoek (1677) ‘Concerning little animal’. Philos Trans R Soc Lond B Biol Sci 370(1666):20140344

    Article  PubMed  PubMed Central  Google Scholar 

  • Lichtman JW, Conchello JA (2005) Fluorescence microscopy. Nat Meth 2:910–919

    Article  CAS  Google Scholar 

  • Mandracchia B, Hua X, Guo C, Son J, Urner T, Jia S (2020) Fast and accurate sCMOS noise correction for fluorescence microscopy. Nat Commun 11(1):1–12

    Article  Google Scholar 

  • Masters BR, So PTC, Gratton E (1997) Multiphoton excitation fluorescence microscopy and spectroscopy of in vivo human skin. Biophys J 72(6): 2405–2412. Bibcode:1997BpJ…72.2405M

    Google Scholar 

  • Mualla F, Aubreville M, Maier A (2018) Microscopy, Chapter 5. In: Maier A, Steidl S, Christlein V et al (eds) Medical imaging systems: an introductory guide [Internet]. Springer, Cham, CH. https://doi.org/10.1007/978-3-319-96520-8_5

    Chapter  Google Scholar 

  • Murphy DB, Davidson MW (2012) Fundamentals of light microscopy and electronic imaging. Wiley, Hoboken, NJ

    Book  Google Scholar 

  • Mutasim DF, Adams BB (2001) Immunofluorescence in dermatology. J Am Acad Dermatol 45:803–822. quiz 22–4

    Article  CAS  PubMed  Google Scholar 

  • Oldenbourg R (2013) Polarized light microscopy: principles and practice. Cold Spring Harb Protoc 11:pdb.top078600. https://doi.org/10.1101/pdb.top078600

    Article  Google Scholar 

  • Pawley JB (ed) (2006) Handbook of biological confocal microscopy, 3rd edn. Springer, Berlin. ISBN 0-387-25921-X

    Google Scholar 

  • Pollock HM, Kazarian GS (2014) Microspectroscopy in the mid-infrared. In: Meyers RA (ed) Encyclopedia of analytical chemistry. Wiley, pp 1–26

    Google Scholar 

  • Rappaz B, Breton B, Shaffer E, Turcatti G (2014) Digital holographic microscopy: a quantitative label-free microscopy technique for phenotypic screening. Comb Chem High Throughput Screen 17(1):80–88

    Article  CAS  PubMed  Google Scholar 

  • Ray SF (2002) Applied photographic optics: lenses and optical systems for photography, film, video, electronic and digital imaging. Focal Press, p 40. ISBN 0-240-51540-4

    Book  Google Scholar 

  • Rust MJ, Bates M, Zhuang X (2006) Sub-diffraction-limit imaging by stochastic optical reconstruction microscopy (STORM). Nat Methods 3:793–795

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Sanderson MJ, Smith I, Parker I, Bootman MD (2014) Fluorescence microscopy. Cold Spring Harb Protoc, p pdb.top071795. https://doi.org/10.1101/pdb.top071795

    Book  Google Scholar 

  • Solem JC (1982) High-intensity x-ray holography: an approach to high-resolution snapshot imaging of biological specimens. Los Alamos National Laboratory Technical Report LA-9508-MS. 83:23581

    Google Scholar 

  • Solem JC (1983) X-ray imaging on biological specimens. Proc Int Conf Lasers 83:635–640

    Google Scholar 

  • The University of Edinburgh (2018) What is microscopy?. The University of Edinburgh. Retrieved June 30, 2022

    Google Scholar 

  • Thomas JL, Rudolph W (2008) Biological microscopy with ultrashort laser pulses. In: Duarte FJ (ed) Tunable laser applications, 2nd edn. CRC Press, Boca Raton, FL, pp 245–280

    Chapter  Google Scholar 

  • Trache A, Meininger GA (2008) Atomic force microscopy (AFM). Current protocols in microbiology, Chapter 2. doi:https://doi.org/10.1002/9780471729259.mc02c02s8

  • Tsien RY Waggoner A (1995) Fluorophores for confocal microscopy. In Pawley JB (ed) Handbook of biological confocal microscopy. Plenum, New York, pp 267–274. ISBN 0-306-44826-2. Retrieved 2008-12-13

    Google Scholar 

  • Van Helden A, Dupré S; van Gent R (2010) The origins of the telescope. Amsterdam University Press, p 24. ISBN 978-90-6984-615-6. Archived from the original on February 15 2017

    Google Scholar 

  • Wäldchen S, Lehmann J, Klein T, Van De Linde S, Sauer M (2015) Light-induced cell damage in live-cell super-resolution microscopy. Sci Rep 5(1):1–12

    Article  Google Scholar 

  • Wayne (2021) Magnification ratio and how to choose the Best macro lens. Retrieved June 30, 2022

    Google Scholar 

  • William Rosenthal (1996) Spectacles and other vision aids: a history and guide to collecting. Norman Publishing, pp 391–392

    Google Scholar 

  • Yao J, Wang LV (2013) Photoacoustic microscopy. Laser Photonics Rev 7(5):1–36

    Article  Google Scholar 

  • Zinselmeyer BH, Dempster J, Wokosin DL, Cannon JJ, Pless R, Parker I, Miller MJ (2009) Chapter 16. Two-photon microscopy and multidimensional analysis of cell dynamics. Methods Enzymol 461:349–378

    Article  CAS  PubMed  Google Scholar 

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Pushpa, N.B., Patra, A., Ravi, K.S. (2023). Advances in Microscopy and Its Applications with Special Reference to Fluorescence Microscope: An Overview. In: Abdel Meguid, E., Mishall, P.L., Nation, H.L., Rea, P.M. (eds) Biomedical Visualisation. Advances in Experimental Medicine and Biology, vol 1406. Springer, Cham. https://doi.org/10.1007/978-3-031-26462-7_1

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