Abstract
The previous chapter described how the confocal approach developed from conventional light microscopy and outlined the basic advantages to be gained by the use of confocal sampling techniques. This chapter will discuss the fundamental considerations that limit the performance of all confocal microscopes. Though at present no commercially available equipment approaches these limits, some simple tests will be described to help the user assess how closely a given instrument performs. Additional information to help the user to operate the confocal microscope in an optimal manner can be found in Chapter 36, “Tutorial on Practical Confocal Microscopy and Use of the Confocal Test Specimen,” which also includes methods for measuring resolution.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Similar content being viewed by others
References
Gelles, J., Schnapp, B.J., Steur, E., and Scheetz, M.P., 1988, Nanometer scale motion analysis of microtubule-based motor enzymes, Proc. EMSA 46:68–69.
Gunter, W.D. Jr., Grant, G.R., and Shaw, S., 1970, Optical devices to increase photocathode quantum efficiency, Appl. Optics 9(2):251–257.
Oldenbourg, R., Terada, H., Tiberio, R., and Inoué, S., 1993, Image sharpness and contrast transfer in coherent confocal microscopy, J. Microsc. 172:31–39.
Paddy, M.R., Hiraoka, Y., Chen, H., Sedat, J.W., and Agard, D.A., 1988, A structural overview of the nucleus as derived from CCD-based 3-D optical microscopy, Proc. EMSA 46:34–35.
Pawley, J.B., 1994, The sources ofnoise in three-dimensional microscopical data sets. In: Three Dimensional Confocal Microscopy: Volume Investigation of Biological Specimens (J. Stevens, ed.), Academic Press, New York, pp. 47–94.
Pawley, J.B., and Centonze, V., 1994, Practical laser-scanning confocal light microscopy: Obtaining optimal performance from your instrument. In: Cell Biology: A Laboratory Handbook, (J.E. Celis, ed.), Academic Press, New York, Vol. 2, pp. 44–64.
Pawley, J.B., Amos, W.B., Dixon, A., and Brelje, T.C., 1993a, Simultaneous, non-interfering, collection of optimal fluorescent and backscattered light signals on the MRC-500/600, Proc. Microsc. Soc. Am. 51:156–157.
Pawley, J.B., Hasko, D., and Cleaver, J., 1993b, A standard test and calibration specimen for confocal microscopy II. In: Proceedings of the 1993 International Conference on Confocal Microscopy and 3-D Image Processing, Sydney, Australia (C.J.R. Sheppard, ed.), p. 35d.
Pawley, J.B., Wright, A.G., and Garrard, C.C., 1993c, Optical enhancement and pulse-counting improve the quality of confocal data. In: Proceedings of the 1993 International Conference on Confocal Microscopy and 3-D Image Processing, Sydney, Australia (C.J.R. Sheppard, ed.), p. 69.
Pollock, H.N., and Chapman, D.S., 1993, Underground records of changing climate, Sci. Am. 68:44–50.
Rose, A., 1948, Television pickup tubes and the problem of noise, Adv. Electron. 1:131.
Sandison, D.R., Piston, D.W., and Webb, W.W., 1994, Background rejection and optimization of signal to noise ratio in confocal microscopy. In: Three-Dimensional Confocal Microscopy: Volume Investigations of Biological Systems (J.K. Stevens, L.R. Mills, and J.E. Trogadis, eds.), Academic Press, New York, pp. 29–48.
Sheppard, J. R., Gu, M., and Roy, M., 1992, Signal-to-noise ratio in confocal microscope systems, J. Microsc. 168:209–218.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 1995 Springer Science+Business Media New York
About this chapter
Cite this chapter
Pawley, J. (1995). Fundamental Limits in Confocal Microscopy. In: Pawley, J.B. (eds) Handbook of Biological Confocal Microscopy. Springer, Boston, MA. https://doi.org/10.1007/978-1-4757-5348-6_2
Download citation
DOI: https://doi.org/10.1007/978-1-4757-5348-6_2
Publisher Name: Springer, Boston, MA
Print ISBN: 978-1-4757-5350-9
Online ISBN: 978-1-4757-5348-6
eBook Packages: Springer Book Archive