Abstract
The previous chapter described how the confocal approach developed from conventional light microscopy and outlined the basic advantages gained by the use of confocal sampling techniques, primarily that the exclusion of light by the confocal pinhole makes it possible to record data from optical sections. 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 well a given instrument performs. Additional information to help the user to operate the confocal microscope in an optimal manner can be found in Chapter 35, “ATutorial on Confocal Microscopy,” and Chapter 36, “Practical Confocal Microscopy.” These also include methods for measuring resolution and other useful parameters.
This is a preview of subscription content, log in via an institution to check access.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
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. Opt. 9:251–257.
Oldenbourg, R., Terada, H., Tiberio, R., Inoué, S., 1993, Image sharpness and contrast transfer in coherent confocal microscopy. J. Microscopy 172:31–39.
Pawley, J.B., 1994, The sources of noise 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., 2000, The 39 steps: Acautionary tale about “quantitative" 3D fluorescence microscopy, BioTechniques, 28:884.
Pawley, J.B., 2002, Limitations on optical sectioning in live-cell confocal microscopy, Scanning, 21:241–246.
Pawley, J.B., and Centonze, V., 1998, 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, pp. 149–169.
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., Blouke, M., and Janesick, J., 1996, The CCDiode: An optimal detector for laser confocal microscopes, Proc. SPIE 2655:125–129.
Pawley, J.B., Hasko, D., and Cleaver, J., 1993, 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 (C.J.R. Sheppard, ed). Sydney, Australia, p. 35.
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., Williams, M., Webb, W.W., 1994, Quantitative comparison of background rejection, signal-to-noise ratio, and resolution in confocal and fullfield laser scanning microscopes. Appl. Opt. 1994: 3576–3588.
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
© 2006 Springer Science+Business Media, LLC
About this chapter
Cite this chapter
Pawley, J.B. (2006). Fundamental Limits in Confocal Microscopy. In: Pawley, J. (eds) Handbook Of Biological Confocal Microscopy. Springer, Boston, MA. https://doi.org/10.1007/978-0-387-45524-2_2
Download citation
DOI: https://doi.org/10.1007/978-0-387-45524-2_2
Publisher Name: Springer, Boston, MA
Print ISBN: 978-0-387-25921-5
Online ISBN: 978-0-387-45524-2
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)