Abstract
Confocal scanning microscopes are particularly attractive by virtue of their enhanced lateral resolution, purely coherent image formation and optical sectioning (Wilson and Sheppard, 1984). It is probably the latter property which is most useful as it gives rise to the ability to image a thick specimen in three-dimensions. This is possible because the optical system images information only from a thin region in the neighbourhood of the focal plane. This permits us to store many image slices in a computer to give a three dimensional data set which describes the object. There are now many sophisticated computer software systems available to display this data in various ways. Obvious examples include the extended focus technique (Wilson and Hamilton, 1982) in which we merely add up (integrate) the images from various depths to provide an image of greatly extended depth of field. We may also produce images where object height is coded as brightness or combine the whole data set to provide an isometric view of the object, Figure 1. It is ;also possible to use false color to label features of interest or, by simple processing, to obtain stereoscopic pairs (van der Voort, 1985). Other forms of image display are discussed in Chapter 10.
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References
Awamura, D., Ode, T. & Tonezawa, M. (1987) Colour laser microscope. SPIED, 765, 53–60.
Corle, T.R., Chou, C.H. & Kino, G.S. (1986) Depth response of confocal optical microscopes. Opt. Lett. 11, 770–772.
Cox, I.J., Sheppard, C.J.R. & Wilson, T. (1982) Super-resolution by confocal fluorescent microscopy. Optik, 60, 391–396.
Cox, I.J. & Sheppard, C.J.R. (1986) Image Capacity and resolution in an optical system. J. Opt. Soc. Am (A), 3, 1152–1158.
Egger, M. D. & Petran, M.(1967) New reflected light microscope for viewing unstained brain and ganglian cells. Science, 157. 305–308.
Hamilton, D.K., and Wilson, T. (1982a) Surface profile measurement using the confocal microscope. J. Appl. Phys. 53, 5320–5322.
Hamilton, D.K., and Wilson, T. (1982b) Three dimensional surface measurement using the confocal scanning microscope. Appl. Phys. B, 27,211–213.
Koester, C.J., (1980) Appl. Opt. 19, 1749.
Mathews, HJ., D. Phil Thesis, (1987) Oxford University.
Schutt, W. (1988) Proc. Conf. on Physical Characterisation of Biological Cells. Rostock GDR.
Sheppard, C.J.R. & Wilson T. (1981a) The theory of the direct-view confocal microscope. J. Microsc. 124, 107–117.
Sheppard, C.J.R. & Wilson, T. (1981b) Effects of high angles of convergence on V(z) in the scanning acoustic microscope. Appl. Phys. Lett. 38, 858–859.
Wilson, T. & Hamilton, D.K. (1982) Dynamic focussing in the scanning microscope. J. Microsc. 128, 139–143.
Wilson, T. & Sheppard, C.J.R. (1984) Theory and Practice of Scanning Optical Microscopy. Academic Press, London.
Wilson, T. (1988) Depth response of scanning microscopes. Optik (in press).
Wilson, T. & Carlini, A.R. (1987) Size of the detector in confocal imaging systems. Opt. Lett. 12, 227–229.
Wilson, T. & Carlini, A.R. (1988a) Three dimensional imaging in confocal imaging systems with finite sized detectors. J. Microsc. 149, 51–66.
Wilson, T. (1989) Optical sectioning in confocal fluorescent microscopes. J. Microsc.
Van der Voort, H.T.M., Brakenhoff, G.J., Valkenburg, J.A.C., Nanninga, N. (1985) Design and use of a computer controlled confocal microscope. Scanning, 7, 66–78.
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© 1990 Plenum Press, New York
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Wilson, T. (1990). The Role of the Pinhole in Confocal Imaging Systems. In: Pawley, J.B. (eds) Handbook of Biological Confocal Microscopy. Springer, Boston, MA. https://doi.org/10.1007/978-1-4615-7133-9_11
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DOI: https://doi.org/10.1007/978-1-4615-7133-9_11
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