Handbook of Biological Confocal Microscopy pp 167-182 | Cite as
The Role of the Pinhole in Confocal Imaging System
Abstract
Confocal scanning microscopes are particularly attractive by virtue of their enhanced lateral resolution, purely coherent image formation (in reflection) and optical sectioning (Wilson and Sheppard, 1984). It is probably the latter property that 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 detects information only from a thin region in the neighborhood of the focal plane. This permits us to store many image slices in a computer to give a three-dimensional (3D) data set that describes the object. Many sophisticated computer software systems are now available to display these 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 in which object height is coded as brightness, or combine the whole data set to provide an isometric view of the object (Fig. 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, et al., 1985).
Keywords
Optical Section Detector Size Coherent Detector Axial Response Pupil FunctionPreview
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References
- Awamura, D., Ode, T., and Tonezawa, M., 1987, Colour laser microscope, SPIED 765:53–60.CrossRefGoogle Scholar
- Corle, T.R., Chou, C.H., and Kino, G.S., 1986, Depth response of confocal optical microscopes, Opt. Lett. 11:770–772.PubMedCrossRefGoogle Scholar
- Cox, I.J., and Sheppard, C. J.R., 1986, Image capacity and resolution in an optical system, J. Opt. Soc. Am (A) 3:1152–1158.CrossRefGoogle Scholar
- Egger, M.D., and Petráň, M., 1967, New reflected light microscope for viewing unstained brain and ganglian cells, Science 157:305–308.PubMedCrossRefGoogle Scholar
- Hamilton, D.K., and Wilson, T., 1982a, Surface profile measurement using the confocal microscope, J. Appl. Phys. 53:5320–5322.CrossRefGoogle Scholar
- Hamilton, D.K., and Wilson, T., 1982b, Three dimensional surface measurement using the confocal scanning microscope, Appl. Phys. B 27:211–213.CrossRefGoogle Scholar
- Juskaitis, R., and Wilson, T., 1992a, Differential confocal scanning microscope using a two mode fibre, Appl. Opt. 31:898–902.PubMedCrossRefGoogle Scholar
- Juskaitis, R., and Wilson, T., 1992b, Imaging in reciprocal fibre-optic based confocal scanning microscopes, Opt. Commun. 92:315–325.CrossRefGoogle Scholar
- Juskaitis, R., and Wilson, T., 1992c, Surface profiling with scanning optical microscopes using two-mode optical fibres, Appl. Opt. 31:4569–4574.PubMedCrossRefGoogle Scholar
- Juskaitis, R., Reinholz, F., and Wilson, T., 1992, Fibre-optic based confocal scanning microscopy with semiconductor laser excitation and detection, Electron. Lett. 28:986–987.CrossRefGoogle Scholar
- Juskaitis, R., Rea, N.P., and Wilson, T., 1993a, Fibre optic based confocal scanning microscopy using laser detection. Opt. Commun. 99:105–113.CrossRefGoogle Scholar
- Juskaitis, R., Wilson, T. and Reinholtz, F., 1993b, Spatial filtering by laser detection in confocal microscopy, Opt. Lett. 18:1135–1137.PubMedCrossRefGoogle Scholar
- Kimura, S., and Wilson, T., 1991, Confocal scanning optical microscopes using single mode fibre for signal detection, Appl. Opt. 30:2143–2150.PubMedCrossRefGoogle Scholar
- Koester, C.J., 1980, A scanning mirror microscope with optical sectioning characteristics: Applications in ophthalmology, Appl. Opt. 19:1749.PubMedCrossRefGoogle Scholar
- Mathews, H.J., 1987, Confocal microscopy, D.Phil thesis, Oxford University, Oxford.Google Scholar
- Schutt, W., 1988, Type III microscopy. In: Proceedings of a Conference on Physical Characterisation of Biological Cells, Rostock, Germany.Google Scholar
- Sheppard, C.J.R., and Wilson T., 1981a, The theory of the direct-view confocal microscope, J. Microsc. 124:107–117.PubMedCrossRefGoogle Scholar
- Sheppard, C.J.R., and Wilson, T., 1981b, Effects of high angles of convergence on V(z) in the scanning acoustic microscope, Appl. Phys. Lett. 38:858–859.CrossRefGoogle Scholar
- van der Voort, H.T.M., Brakenhoff, G. J., Valkenburg, J.A.C., and Nanninga, N., 1985, Design and use of a computer controlled confocal microscope, Scanning 7:66–78.CrossRefGoogle Scholar
- Wilson, T., 1989, Optical sectioning in confocal fluorescent microscopes, J. Microsc. 154:143–156.CrossRefGoogle Scholar
- Wilson, T., 1993a, Image formation in two-mode fibre based confocal microscopy, J. Opt. Soc. Am. 10:1535–1543.CrossRefGoogle Scholar
- Wilson, T., 1993b, Fluorescence imaging modes in fibre-optic based confocal scanning microscopes, Opt. Commun. 96:133–141.CrossRefGoogle Scholar
- Wilson, T., and Carlini, A.R., 1987, Size of the detector in confocal imaging systems, Opt. Lett. 12:227–229.PubMedCrossRefGoogle Scholar
- Wilson, T., and Carlini, A.R., 1988, Three dimensional imaging in confocal imaging systems with finite-sized detectors, J. Microsc. 149:51–66.CrossRefGoogle Scholar
- Wilson, T., and Carlini, A.R., 1989, The effect of aberrations on the axial response of confocal imaging-systems, J. Microsc. 154:243–256.CrossRefGoogle Scholar
- Wilson, T., and Hamilton, D.K., 1982, Dynamic focussing in the scanning microscope, J. Microsc. 128:139–143.CrossRefGoogle Scholar
- Wilson, T., and Hamilton, D.K., 1984, Differential confocal scanning micros copy, Opt. Acta 31:453–465.CrossRefGoogle Scholar
- Wilson, T., and Hewlett, S. J., 1991, Optical sectioning strength of the direct-view microscope employing finite-sized pin-hole arrays, J. Microsc. 163:131–150.CrossRefGoogle Scholar
- Wilson, T., and Sheppard, C.J.R., 1984, Theory and Practice of Scanning Optical Microscopy, Academic Press, London.Google Scholar