Bibliography of Confocal Microscopes

  • Robert H. Webb


This is a selected bibliography, as of early 1994, of accessible papers on confocal microscopy. Not included are conference reports and other documents not likely to be available in most technical libraries. I have tried to include a note as to the content— often taken from the abstract.


Confocal Microscopy Confocal Scanning Fluorescence Correlation Spectroscopy Spherical Aberration Optical Transfer Function 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.


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Books and Review Articles

  1. Baxes, G.A., 1984, Digital Image Processing: A Practical Primer, Prentice-Hall, Englewood Cliffs, New Jersey. Also (1988) from Cascade Press, P.O. Box 27631, Denver, CO 80227. Truly a “practical primer.”Google Scholar
  2. Boyde, A., 1994, Bibliography on confocal microscopy and its applications, Scanning 16:33–56. Some overlap with this chapter, but not a direct mapping.Google Scholar
  3. Boyde, A., Jones, S.J., Taylor, M.L., Wolfe, L.A., and Watson, T.F., 1990, Fluorescence in the tandem scanning microscope, J. Microsc. 157:39–49. TSMs and CSMs.PubMedCrossRefGoogle Scholar
  4. Brelje, T.C., Wessendorf, M.W., and Sorenson, R.L., 1993, Multi-color laser scanning confocal immunofluorescence microscopy: Practical application and limitations, Methods Cell Biol. 38:98–182. A review of the dyes and the techniques for using them. Very specific. Complete.Google Scholar
  5. Cagnet, M., Françon, M., and Thrierr, J.C., 1962, Atlas of Optical Phenomena, Springer-Verlag, Berlin. See p. 23 for a beautiful through-focus display.Google Scholar
  6. Castleman, K.R., 1979, Digital Image Processing, Prentice-Hall, Englewood Cliffs, New Jersey. A standard reference on this subject.Google Scholar
  7. Inoué, S., 1986, Video Microscopy, Plenum Press, New York. A very complete basic book.Google Scholar
  8. Inoué, S, and Oldenbourg, R., 1994, Optical instruments: Microscopes. In: Handbook of Optics (M. Bass, ed.), 2nd ed., Vol. 2, Chap. 17, McGraw-Hill, New York. An up-to-date general reference.Google Scholar
  9. Kino, G.S., and Corle, T.R., 1989, Confocal scanning optical microscopy, Phys. Today 42:55–62. A review article for nonspecialist physicists.CrossRefGoogle Scholar
  10. Oppenheim, A.V., Willsky, A.S., and Young, LT., 1983, Signals and Systems, Prentice-Hall, Englewood Cliffs, New Jersey. Chapter 8 describes sampling theory very well.Google Scholar
  11. Pawley, J.B., 1991, Fundamental and practical limits in confocal light microscopy, Scanning 13:184–198. A general review of some of the choices we all make, with attention to photobleaching problems, optimization of optics, and pinholes.CrossRefGoogle Scholar
  12. Petran, M., Hadravsky, M., and Boyde, A., 1985 The tandem scanning reflected light microscope, Scanning 7:97–108. Summarizes the TSM.CrossRefGoogle Scholar
  13. Pratt, W.K., 1978, Digital Image Processing, Wiley, New York. A standard reference in this field.Google Scholar
  14. Shotton, D., ed., 1993, Electronic Light Microscopy—Techniques in Modem Biomedical Microscopy, Wiley-Liss, New York. Many of the usual suspects have contributed chapters, with some further chapters on display, video and sample preparation. A good companion to this volume.Google Scholar
  15. Slater, E.M., and Slater, H.S., 1993, Light and Electron Microscopy, Cambridge University Press, London. A fine general book on microscopes.Google Scholar
  16. Stevens, J.K., Mills, L.R., and Trogadis, J., eds., 1993, Three-Dimensional Confocal Microscopy, Academic Press, New York. A recent addition.Google Scholar
  17. Webb, R.H., 1991, Confocal microscopes, Optics & Photonics News 2:8–13. A review for optical scientists who do not specialize in microscopes.CrossRefGoogle Scholar
  18. White, J.G., and Amos, W.B., 1987, Confocal microscopy comes of age, Nature 328:183. General article by two of the early users of the CSM.CrossRefGoogle Scholar
  19. Wilson, T., 1980, Imaging properties and applications of scanning optical microscopes, Appl. Phys. (Germany) 22:119–128. A review paper, with 57 references.CrossRefGoogle Scholar
  20. Wilson, T., 1985, Scanning optical microscopy, Scanning 7:79–87. Summary article.CrossRefGoogle Scholar
  21. Wilson, T., ed., 1990, Confocal Microscopy, Academic Press, New York. Chapters by many of the usual folk. Much of the thinking of the 1980s summarized and updated.Google Scholar
  22. Wilson, T., and Sheppard, C.J.R., 1984, Theory and Practice ofScanning Optical Microscopy, Academic Press, New York. Reprints some of their early papers.Google Scholar

Historical Interest

  1. Some of the original articles, mostly now superseded by later work—often of the same authors. I have tried to avoid the “gee whiz” articles.Google Scholar
  2. Amos, W.B., White, J.G., and Fordham, M., 1987, Use of confocal imaging in the study of biological structures, Appl. Optics 26:3239–3243. A good general review, and an early description of the MRC (Biorad) microscope.CrossRefGoogle Scholar
  3. Åslund, N., Liljeborg, A., Forsgren, P.-O., and Wahlsten, S., 1987, Three-dimensional digital microscopy using the PHOIBOS scanner, Scanning 9:227–235. Consecutive optical sections to generate digital three-dimensional microscopy.CrossRefGoogle Scholar
  4. Baer, S.C., 1970, Optical apparatus providing focal-plane specific information, U.S. Patent 3,547,512. An early CSM idea, largely ignored for 17 years.Google Scholar
  5. Brakenhoff, G.J., Blom, P., and Barends, P., 1979, Confocal scanning light microscopy with high aperture immersion lenses, J. Microsc. 117:219–232. For point objects the theoretically expected factor of 1.4 can be realized. A further improvement by a factor of 1.25 after apodization with an annular aperture.CrossRefGoogle Scholar
  6. Brakenhoff, G.J., van der Voort, H.T.M., van Spronsen, E.A., Linnemans, W.A.M., and Nanninga, N., 1985, Three-dimensional chromatin distribution in neuroblastoma nuclei shown by confocal scanning laser microscopy, Nature 317:748–749. An early use of optical sectioning and higher resolution for studying the three-dimensional morphology of biological structures.PubMedCrossRefGoogle Scholar
  7. Chou, C.-H., and Kino, G.S., 1987, The evaluation of V(z) in a type II reflection microscope, IEEE Trans. Ultrason. Ferroelectr. Freq. Control 34:341–345. Theory: V(z) of an acoustic microscope, nonparaxial and finite pinhole and the asymmetry of the V(z) curve for a perfect reflector.PubMedCrossRefGoogle Scholar
  8. Davidovits, P., and Egger, M.D., 1971, Scanning laser microscope for biological investigations, Appl. Optics 10:1615–1619. Also: Davidovits, P., and Egger, M.D., 1969, Scanning laser microscope. Nature 223:831. Two seminal papers of historical interest.CrossRefGoogle Scholar
  9. Egger, M.D., and Petráň, M., 1967, New reflected-light microscope for viewing unstained brain and ganglion cells, Science 157:305–307. A largely unnoticed description of the TSM, but it was all therePubMedCrossRefGoogle Scholar
  10. Hamilton, D.K., and Wilson, T., 1982, Three-dimensional surface measurement using the confocal scanning microscope, Appl. Phys. B 27:211–213. Early profilometry on semiconductors, to 0.1 p. m.CrossRefGoogle Scholar
  11. Hamilton, D.K., Wilson, T., and Sheppard, C.J.R., 1981, Experimental observations of the depth-discrimination properties of scanning microscopes, Opt. Lett. 6:625–626. Optical sectioning.PubMedCrossRefGoogle Scholar
  12. Maurice, D.M., 1973, A scanning slit optical microscope, Invest. Ophthalmol. 13:1033–1037. A pioneer paper describing an early form of confocal microscopy for imaging layers in the cornea of the eye. This system used a scanning slit 3 μm wide to give depth definition, and scanning was carried out by moving a photographic film and the specimen in opposite directions. High-quality images of the cornea were obtained, which took about 20 minutes to form.Google Scholar
  13. Minsky, M., 1988, Memoir on inventing the CSM, Scanning 10:128–138. A valuable historical document, and enjoyable reading. Minsky’s patent ran out before the world was ready for the idea, but his early ideas have all proved out well. See Minsky, M., 1961, Microscopy apparatus, U.S. Patent 3,013,467.CrossRefGoogle Scholar
  14. Petráň, M., and Hadravsky, M., 1967, Method and arrangement for improving the resolving power and contrast, U.S. Patent 3,517,980, filed 4–12-67, granted 30–6–70.Google Scholar
  15. Petráň, M., and Hadravsky, M., 1968, Tandem-scanning reflected-light microscope, J. Opt. Soc. Am. 58:661–664. Source paper on TSM. See also Petráň, M., Hadravsky, M., Benes, J., Kucera, R., and Boy de, A., 1985, The tandem scanning reflected light microscope. Part 1—The principle, and its design, Proc. R. Microsc. Soc. 20:125–129.CrossRefGoogle Scholar
  16. Ploem, J.S., 1987, Laser scanning fluorescence microscopy, Appl. Optics 26 3226–3231. Another general description of CSM.CrossRefGoogle Scholar
  17. Sheppard, C.J.R., and Choudhury, A., 1977, Image formation in the scanning microscope, Opt. Acta 24:1051–1073. Fourier imaging in microscopes of type 1 (conventional) and type 2 (confocal). Single- and two-point resolution, response to a straight edge, annular pupil functions.CrossRefGoogle Scholar
  18. Sheppard, C.J.R., and Wilson, T., 1978, Depth of field in the scanning microscope, Opt. Lett. 3:115–117. Various definitions of depth of field in the microscope are discussed.PubMedCrossRefGoogle Scholar
  19. Toraldo di Francia, G., 1955, Resolving power and information, J. Opt. Soc. Am. 45:497–501. Two-point resolution is impossible unless the observer has a priori an infinite amount of information about the object.CrossRefGoogle Scholar
  20. Welford, W.T., 1972, On the relationship between the modes of image formation in scanning microscopy and conventional microscopy, J. Microsc. 96:105–107.PubMedCrossRefGoogle Scholar
  21. White, J.G., Amos, W.B., and Fordham, M., 1987, An evaluation of confocal versus conventional imaging of biological structures by fluorescence light microscopy, J. Cell Biol. 105–41-48. An early general article which helped introduce the technique.PubMedCrossRefGoogle Scholar
  22. Wilke, V., 1985, Optical scanning microscopy—The laser scan microscope, Scanning 7:88–96. The Zeiss CSM.CrossRefGoogle Scholar
  23. Wilson, T., and Hamilton, D.K., 1982, Dynamic focusing in the confocal scanning microscopes, J. Microsc. 128:139–143. Image built up as each section of the object passes through the focal plane.CrossRefGoogle Scholar

Theory (Mostly)

  1. Bertero, B., De Mol, C., and Pike, E.R., 1987, Analytic inversion formula for confocal scanning microscopy, J. Opt. Soc. Am. A 4:1748–1750. A simple analytic expression for the inverse problem in CSMs.CrossRefGoogle Scholar
  2. Carlsson, K., 1991, The influence of specimen refractive index, detector signal integration, and non-uniform scan speed on the imaging properties in confocal microscopy, J. Microsc. 163:2, 167–178. Index mismatch causes spherical aberrations, which affect axial resolution most.CrossRefGoogle Scholar
  3. Cogswell, C.J., and Sheppard, C.J.R., 1992, Confocal differential interference contrast (DIC) microscopy: Including a theoretical analysis of conventional and confocal DIC imaging, J. Microsc. 165:81–101. DIC with a CSM is compared to existing confocal differential phase contrast (DPC) techniques and to conventional Nomarski DIC. A theoretical treatment of DIC imaging is presented, which takes into account vignetting by the lens pupils.CrossRefGoogle Scholar
  4. Corle, T.R., and Kino, G.S., 1990, Differential interference contrast imaging on a real time confocal scanning optical microscope, Appl. Optics 29:3769–3774. The advantage of DIC in a CSM is that both the height and width of an edge can be measured without ambiguity, even if the edge is taller than half a wavelength.CrossRefGoogle Scholar
  5. Cox, I.J., and Sheppard, C.J.R., 1986, Information capacity and resolution in an optical system, J. Opt. Soc. Am. 3:1152–1158. A nonstandard approach that gives useful general results without major math.CrossRefGoogle Scholar
  6. Cox, I.J., Sheppard, C.J.R., and Wilson, T., 1981, Improvement in resolution by nearly confocal microscopy: The theory of the direct-view confocal microscope, J. Microsc. 124:107–117. Resolution increased by offsetting the pinhole. Dark-field conditions are produced with the pinhole over the first dark ring in the Airy disk. Theory for conventional and scanning microscopes, partial coherence and TSMs.CrossRefGoogle Scholar
  7. Drazic, V., 1992, Three-dimensional transfer function of coherent confocal microscopes with extended source and detector, J. Mod. Opt. 39:1777–1790. The effect of finite size of the source and detector on the three-dimensional transfer function of an incident light coherent CSM.CrossRefGoogle Scholar
  8. Gu, M., and Sheppard, C.J.R., 1992, Confocal fluorescent microscopy with a finite-sized circular detector, J. Opt. Soc. Am. A, Opt. Image Sci. 9:151–153. OTF has negative values when the detector radius exceeds certa in magnitudes.CrossRefGoogle Scholar
  9. Gu, M., and Sheppard, C.J.R., 1992, Three-dimensional optical transfer function in a fiber-optical confocal fluorescence microscope using annular lenses, J. Opt. Soc. Am. A, Opt. Image Sci. 9:1991–1999. Annular lenses in a system with optical fibers can result in improved resolution in both transverse and axial directions.CrossRefGoogle Scholar
  10. Gu, M., and Sheppard, C.J.R., 1992, Effects of defocus and primary spherical aberration on three-dimensional coherent transfer functions in confocal microscopes, Appl. Optics 31:2541–2549. Three-dimensional confocal imaging is strongly degraded if the amount of aberration is larger than a quarter wavelength. Spherical aberration compensated by defocus.CrossRefGoogle Scholar
  11. Hamilton, D.K., and Wilson, T., 1984, Two-dimensional phase imaging in the scanning optical microscope, Appl. Optics 23:348–352. Early split-detector phase imaging.CrossRefGoogle Scholar
  12. Hegedus, Z.S., 1985, Annular pupil arrays. Application to confocal scanning, Opt. Acta 32:815–826. Radially symmetrical pupil masks with a continuously varying transmittance can be made by vacuum deposition or photographic integration, or a binary mask of a concentric array of annuli.CrossRefGoogle Scholar
  13. Hell, S., Reiner, G., Cremer, C., and Stelzer, E.H.K., 1993, Aberrations in confocal fluorescence microscopy induced by mismatches in refractive index, J. Microsc. 169:391–405. An extensive and readable analysis of this frequent topic. Scaling factors for correction are given.CrossRefGoogle Scholar
  14. Hobbs, P.C.D., and Kino, G.S., 1990, Generalizing the confocal microscope via heterodyne interferometry and digital filtering, J. Microsc. 160:245–264. A true heterodyne CSM, yielding both phase and amplitude.CrossRefGoogle Scholar
  15. Inoué, S., 1989, Imaging of unresolved objects, superresolution, and precision of distance measurement with video microscopy, Methods Cell Biol. 30:85–112.PubMedCrossRefGoogle Scholar
  16. Mendez, E.R., 1986, Speckle contrast variation in the CSM. Hard-edged apertures, Opt. Acta 33:269–278. Speckle contrast variation as a function of defocus, and the statistical properties of random diffusing objects.CrossRefGoogle Scholar
  17. Ooki, H., and Iwasaki, J., 1991, A novel type of laser scanning microscope: Theoretical considerations, Opt. Commun. 85:177–182. Differential interference contrast by means of mode interference in a waveguide device. Phase and amplitude separately.CrossRefGoogle Scholar
  18. Sandison, D.R., and Webb, W.W., 1994, Background rejection and signal-to-noise optimization in the confocal and alternative fluorescence microscopes, Applied Optics 33:603. A complete analysis of signal, background and noise in the family of confocal microscopes.PubMedCrossRefGoogle Scholar
  19. Sheppard, C.J.R., 1988, Aberrations in high aperture conventional and confocal imaging systems, Appl. Optics 27:4782–4786. In an aberration function for high numerical aperture the effects on the defocus signal of a confocal imaging system of aberrations, high aperture, finite Fresnel number, system configuration, and surface tilt are discussed.CrossRefGoogle Scholar
  20. Sheppard, C.J.R., and Cogswell, C.J., 1990, Three-dimensional image formation in confocal microscopy, J. Microsc. 159:179–194. 3D imaging in terms of 3D transfer functions.CrossRefGoogle Scholar
  21. Sheppard, C.J.R., and Wilson, T., 1981, The theory of the direct-view confocal microscope, J. Microsc. 124:107–117. Theory embracing conventional microscopes with partially coherent source and scanning microscopes with partially coherent effective source and detector, including confocal microscopes and the TSM of Petráň.PubMedCrossRefGoogle Scholar
  22. Sheppard, C.J.R., and Wilson, T., 1986, Reciprocity and equivalence in scanning microscopes, J. Opt. Soc. Am. A 3:755–756. The principle of reciprocity and methods of Fourier optics in conventional and scanning microscopes: their behavior is identical even for objects thick enough for multiple scattering to occur, provided that there is no inelastic scattering or birefringence present.CrossRefGoogle Scholar
  23. Sheppard, C.J.R., Cogswell, C.J., and Gu, M., 1991, Signal strength and noise in confocal microscopy: Factors influencing selection of an optimum detector aperture, Scanning 13:233–240.CrossRefGoogle Scholar
  24. van der Voort, H.T.M., and Brakenhoff, G.J., 1990, 3-D image formation in high-aperture fluorescence confocal microscopy: A numerical analysis, J. Microsc. 158:43–54. Electromagnetic diffraction theory of the field near focus as developed by Richards and Wolf is used to compute the optical properties of the model.CrossRefGoogle Scholar
  25. Visser, T.D., Groen, F.C.A., and Brakenhoff, G.J., 1991, Absorption and scattering correction in fluorescence confocal microscopy, J. Microsc. 163:189–200. With one space-dependent extinction coefficient, the total attenuation process can be calculated to the deeper layers.CrossRefGoogle Scholar
  26. Wilson, T., 1991, Comment on “Image formation in a superresolution phase conjugate scanning microscope,” Appl. Phys. Lett. 58:314. Johnson, Cathey, and Mao, 1989, Appl. Phys. Lett. 55:1707, proposed using a phase conjugate mirror to exhibit superresolution. A phase conjugate mirror is not strictly necessary and a similar, or even enhanced, effect may be obtained by processing the image from a standard confocal microscope in an extremely simple way.CrossRefGoogle Scholar
  27. Wilson, T., and Carlini, A.R., 1987, Size of the detector in confocal imaging systems, Opt. Lett. 12:227–229. Original of many familiar figures.PubMedCrossRefGoogle Scholar
  28. Wilson, T., Carlini, A.R., 1989, The effect of aberrations on the axial response of confocal imaging systems, J. Microsc. 154:243–256. Aberrations are diminished by smaller pinholes.CrossRefGoogle Scholar


  1. Aikens, R.S., Agard, D.A., and Sedat, J.W., 1989, Solid state imagers for microscopy, Methods Cell Biol. 29:291–313. A complete discussion of the detector of choice in many disk-scanning and wide-field microscopes.PubMedCrossRefGoogle Scholar
  2. Art, J.J., and Goodman, M.B., 1993, Rapid scanning confocal microscopy, Methods Cell Biol. 38. Includes the NORAN Odyssey instrument.Google Scholar
  3. Benedetti, P.A., Evangelista, V., Guidarini, D., and Vestri, S., 1992, Confocal-line microscopy, J. Microsc. 165:119–129. Confocal in one dimension, no moving parts. Linear imagers permit transmission, reflection, and emission images simultaneously.PubMedCrossRefGoogle Scholar
  4. Bliton, C., Lechleiter, J., and Clapham, D.E., 1993, Optical modifications enabling simultaneous confocal imaging with dyes excited by ultra-violet and visible-wavelength light, J. Microsc. 169:15–26. Modifications to a BioRad MRC 600.CrossRefGoogle Scholar
  5. Boyde, A., Xiao, G.Q., Corle, T., Watson, T.F., and Kino, G.S., 1990, An evaluation of unilateral TSM for biological applications, Scanning 12:273–279. Two designs of tandem scanning reflected light microscopes, due to Petráň and to Kino.CrossRefGoogle Scholar
  6. Brakenhoff, G.J., and Visscher, K., 1992, Confocal imaging with bilateral scanning and array detectors, J. Microsc. 165:139–146. A hybrid CSM/TSM using an array detector such as a CCD for confocal image collection and a double-sided scanning mirror to scan and collect data. This is the scheme implemented by Meridian.CrossRefGoogle Scholar
  7. Brakenhoff, G.J., and Visscher, K., 1993, Imaging modes for bilateral confocal scanning microscopy, J. Microsc. 171:17–26. Details of the double-sided mirror rescanning system. The remitted light traverses the confocal pinhole to yet another scanning mirror (the second side of the original one) to be spread again into an image viewable by eye or camera. See the Koester papers for earlier implementations. This is the scheme implemented by Meridian.CrossRefGoogle Scholar
  8. Buurman, E.P., Sanders, R., Draaijer, A., Gerritsen, H.C., Van Veer, J.J.F., Haupt, P.M., and Levine, Y.K., 1992, Fluorescence lifetime imaging using a confocal laser scanning microscope, Scanning 14:155–159. Fluorescence lifetime imaging method in a CSM uses a low-power CW argon laser chopped to 25-MHz nanosecond pulses. Time-gated detection measures the lifetime of a pixel in 40 μ sec.CrossRefGoogle Scholar
  9. Carlsson, K., and Åslund, N., 1987 Confocal imaging or 3-D digital microscopy, Appl. Optics 26:3232–3238. This is the Phoibos instrument of Sarastro, now of Molecular Dynamics.CrossRefGoogle Scholar
  10. Cogswell, C.J., Hamilton, D.K., and Sheppard, C.J.R., 1992, Colour confocal reflection microscopy using red, green and blue lasers, J. Microsc. 165:103–117. Lasers: HeNe (633 nm); NdYAG (532 nm); HeCd (442 nm) and three photomultiplier detectors.CrossRefGoogle Scholar
  11. Dixon, A.E., Damaskinos, S., and Atkinson, M.R., 1991, A scanning confocal microscope for transmission and reflection imaging, Nature 351:551–553. The optical slices in transmission do not change intensity with depth.CrossRefGoogle Scholar
  12. Dixon, A.E., Damaskinos, S., and Atkinson, M.R., 1991, Transmission and double-reflection scanning stage confocal microscope, Scanning 13:299–306. A stage-scanning transmitted- and reflected-light laser microscope is described.CrossRefGoogle Scholar
  13. Entwistle, A., and Noble, M., 1992, The quantification of fluorescent emission from biological samples using analysis of polarization, J. Microsc. 165:347–365. Analysis of fluorescence depolarization can identify regions in which fluorophore concentration exceeds the range of linear fluorescent emission.PubMedCrossRefGoogle Scholar
  14. Fricker, M.D., and White, N.S., 1992, Wavelength considerations in confocal microscopy of botanical specimens, J. Microsc. 166:29–42. Some of the problems associated with multiple wavelengths. CSMs make severe demands on achromatized optics.CrossRefGoogle Scholar
  15. Glass, M., and Dabbs, T., 1991, The experimental effect of detector size on confocal lateral resolution, J. Microsc. 164:153–158. The discrepancy between data and theory attributed to apodization from multielement thick lenses and a nonideal, truncated Gaussian beam profile.CrossRefGoogle Scholar
  16. Hamilton, D.K., and Sheppard, C.J.R., 1986, Interferometric measurements of the complex amplitude of the defocus signal V(z) in the confocal scanning optical microscope, J. Appl. Phys. 60:2708–2712. A confocal interference microscope with electro-optic phase modulator makes simultaneous measurements of the in-phase and quadrature components of the confocal signal as a reflecting surface is scanned axially, the so-called V(z) response.CrossRefGoogle Scholar
  17. Hansen, E.W., Allen, R.D., Strohbehn, J.W., Chaffee, M.A., Farrington, D.L., Murray, W.J., Pillsbury, T.A., and Riley, M.F., 1985, Laser scanning phase modulation microscope, J. Microsc. 140:371–381. Quantitative polarized light imaging with a phase modulation feedback loop for precise measurement of birefringence.PubMedCrossRefGoogle Scholar
  18. Hell, S., Witting, S., Schickfus, M.V., and Neiger, M., 1991, A confocal beam scanning white-light microscope, J. Microsc. 163:179–187. CSM with a continuous Xe short-arc lamp operating in the visible spectrum. Resolution of the white-light microscope is equivalent to that of the scanning laser microscope, without artifacts caused by interference.CrossRefGoogle Scholar
  19. Highett, M.I., Rawlins, D.J., and Shaw, P.J., 1993, Different patterns of rDNA distribution in Pisum sativum nucleoli correlate with different levels of nucleolar activity, J. Cell Sci. 104:843–852. Compares CSM and decon-volution of wide-field images. Both together are best.Google Scholar
  20. Hong Qian, and Elson, E.L., 1991, Analysis of confocal laser-microscope optics for 3-D fluorescence correlation spectroscopy, Appl. Optics 30:1185–1195. Quantitative fluorescence correlation spectroscopy and fluorescence photobleaching recovery measurements.CrossRefGoogle Scholar
  21. Horikawa, Y., Yamamoto, M., andDosaka, S., 1987, Laser scanning microscope: Differential phase images, J. Microsc. 148:1–10. A TV-rate acousto-op-tic deflector laser scanning microscope for differential phase contrast images using the split-detector technique.PubMedCrossRefGoogle Scholar
  22. Janssen, G.C.A.M., Rousseeuw, B.A.C., and van der Voort, H.T.M., 1987, Test pattern for fluorescence microscopy, Rev. Sci. Instrum. 58:598–599. Fluorescent test pattern with submicron dimensions. Measurements of the transfer function of the CSM.CrossRefGoogle Scholar
  23. Mansfield, S.M., and Kino, G.S., 1990, Solid immersion microscope, Appl. Phys. Lett. 57:2615–2616. A real-time optical microscope with the liquid replaced by a solid lens of high-refractive-index material.CrossRefGoogle Scholar
  24. Marsman, H.J.B., Stricker, R., Wijnaendts van Resandt, R.W., Brakenhoff, G.J., and Blom, P. 1983, Mechanical scan system for microscopic applications, Rev. Sci. Instrum. 54:1047–1052. A high-speed mechanical scanning stage for high-resolution confocal UV microscopy.CrossRefGoogle Scholar
  25. Matthews, H.J., Hamilton, D.K., and Sheppard, C.J.R., 1989, Aberration measurement by confocal interferometry, J. Mod. Opt. 36:233–280. Aberrations and apodization of microscope objectives measured from the defocus signal in a confocal interference microscope system. Phase distortions can be measured to approximately 1/100, and quantitative information is given about the imaging performance of the lenses in situ in the optical system.CrossRefGoogle Scholar
  26. Oldenbourg, R., Terada, H., Tiberio, R., and Inoué, S., 1993, Image sharpness and contrast transfer in coherent confocal microscopy, J. Microsc., 172:31–39. How to measure the resolution of your CSM.PubMedCrossRefGoogle Scholar
  27. See, C.W., and Iravani, M.V., 1988, Differential amplitude scanning optical microscope: Theory and applications, Appl. Optics 27:2786–2792. Differential means two adjacent spots, to catch small changes of height or index. This is a dynamic range stretcher for those quantities.CrossRefGoogle Scholar
  28. Shack, R.V., Bartels, P.H., Buchroeder, R.A., Shoemaker, R.L., Hillman, D.W., and Vukobratovich, D., 1987, Design for a fast fluorescence laser scanning microscope, Anal. Quant. Cytol. Histol. 9:509–520. Discussion of the design consideration of the principal components, including the optical elements.PubMedGoogle Scholar
  29. Shaw, P.J., 1990, Three-dimensional optical microscopy using tilted views, J. Microsc. 158:165–172. Multiple tilted views increase the resolution in z. This technique was applied to metaphase chromosomes from intact embryos of Drosophila melanogaster. As determined from significant intensity in the Fourier transform, the resolution of the final reconstruction was about 0.25 μm in x andy, and 0.4 μm in z.PubMedCrossRefGoogle Scholar
  30. Shaw, P.J., and Rawlins, D.J., 1991, The point-spread function of a confocal microscope: Its measurement and use in deconvolution of 3-D data, J. Microsc. 163:151–165. PSF for an MRC-500 CSM using subresolution fluorescent beads. The resulting optical transfer functions were used in an iterative, constrained deconvolution procedure to get three-dimensional confocal data sets from a biological specimen.CrossRefGoogle Scholar
  31. Sheppard, C.J.R., and Cogswell, C.J., 1990, Confocal microscopy with detector arrays, J. Mod. Opt. 37:267–279. Imaging in a scanning optical microscope with a detector consisting of an array of rings is considered. It is found that both transverse and axial resolution can be increased simultaneously.CrossRefGoogle Scholar
  32. Sheppard, C.J.R., and Gu, M., 1991, Aberration compensation in confocal microscopy, Appl. Optics 30:3563–3568. Spherical aberration due to focusing deep within the specimen can be compensated by altering the effective tube length.CrossRefGoogle Scholar
  33. Sheppard, C.J.R., and Gu, M., 1992, Axial imaging through an aberrating layer of water in confocal microscopy, Opt. Commun. 88:180–190. The axial response modeled by a series of signals reflected from different interfaces. Altering the effective tube length for an optimum axial response, experimental results are well in agreement with theoretical predictions.CrossRefGoogle Scholar
  34. Sheppard, C.J.R., Hamilton, D.K., and Cox, I.J., 1982, Optical microscopy with extended depth of field: Observation of optical signatures of materials, Appl. Phys. Lett. 41:604–606. Depth of field may be extended while high-resolution, diffraction-limited imaging is retained. The technique is similar to one already used in acoustic microscopy.CrossRefGoogle Scholar
  35. Shoemaker, R.L., Bartels, P.H., Hillman, D.W., Jonas, J., Kessler, D., Shack, R.V., and Vukobratovich, D., 1982, An ultrafast laser scanner microscope for digital image analysis (cytology application), IEEE Trans. Biomed. Eng. BME-29:82–91. An ultrafast laser scanner microscope to make high-resolution cell analysis practical at data rates comparable to flow cytometry.PubMedCrossRefGoogle Scholar
  36. Stelzer, E.H.K., Marsman, H.J.B., and Wijnaendts van Resandt, R.W., 1986, A setup for a confocal scanning laser interference microscope, Optikl3:30–33. The interference mode installed on a CSM is compared with that on a conventional microscope and high quality images are presented.Google Scholar
  37. Szarowski, D.H., Barnard, D.P., Smith, K.L., Swann, J.W., Holmes, K.V., and Turner, J.N., 1990, Confocal laser-scanned microscopy: Analog signal processing, Scanning 12:265–272. Analog signal processing to adjust dynamic range in a BioRad MRC-500 CSM.CrossRefGoogle Scholar
  38. Szarowski, D.H., Smith, K.L., Herchenroder, A., Matuszek, G., Swann, J.W., and Turner, J.N., 1992, Optimized reflection imaging in laser confocal microscopy and its application to neurobiology: Modifications to the BioRad MRC-500, Scanning 14:104–111. Use of polarization components to control stray light.CrossRefGoogle Scholar
  39. Tsien, R.Y., 1989, Fluorescent probes of cell signaling, Annu. Rev. Neurosci. 12:227–253. A general review of the dyes CSM uses.PubMedCrossRefGoogle Scholar
  40. Tsien, R.Y., and Poenie, M., 1986, Fluorescence ratio imaging: A new window into intracellular ionic signaling, Trends Biochem. Sci. 11:450–455. Describes one of the major tools used in CSM.CrossRefGoogle Scholar
  41. van der Oord, C.J.R., Gerritsen, H.C., Levine, Y.K., Myring, W.J., Jones, G.R., and Munro, LH., 1992, Synchrotron radiation as a light source in confocal microscopy, Rev. Sci. Instrum. 63:632–633. A CSM using a synchrotron as light source from 200 nm up to 700 nm. Using 325-nm laser light, it is shown that the lateral resolution is about 125 nm, and the axial resolution better than 250 nm.CrossRefGoogle Scholar
  42. Van Oostveldt, P., and Bauwens, S., 1990, Quantitative fluorescence in confocal microscopy. The effect of the detection pinhole aperture on the re-absorption and inner filter phenomena, J. Microsc. 158:121–132. The numerical aperture of the objective does not affect the integrated fluorescence intensity and the integrated absorbance in Feulgen-stained pigeon erythrocyte nuclei hydrolyzed for different periods of time and stained at different dye concentrations.CrossRefGoogle Scholar
  43. Webb, R.H., and Hughes, G.W., 1993, Detectors for scanning video imagers, Appl. Optics 32:6227–6235. Avalanche photodiodes are superior to photomultiplier tubes when scan rates are in the video range.CrossRefGoogle Scholar
  44. Wilson, T., 1990, The role of detector geometry in confocal imaging, J. Microsc. 158:133–144. Point detectors, slit detectors, detector arrays, and finite-sized detectors.CrossRefGoogle Scholar
  45. Wilson, T., and Carlini, A.R., 1988, Effect of detector displacement in confocal imaging systems, Appl. Opt. 27:3791–3799. In certain cases a degree of detector offset may be used to advantage in determining the position of an edge.PubMedCrossRefGoogle Scholar
  46. Wilson, T., and Hewlett, S.J., 1990, Imaging in scanning microscopes with slit-shaped detectors, J. Microsc. 160:115–139. An important addition to Wilson’s opera, spelling out the variations for slits. Not particularly reader-friendly.PubMedCrossRefGoogle Scholar
  47. Wilson, T., Carlini, A.R., and Sheppard, C.J.R., 1985, Phase contrast microscopy by nearly full illumination, Optik 70:166–169. A simple phase contrast technique where the two lenses in a scanning microscope are not equal is equally applicable to conventional and CSMs and is demonstrated experimentally.Google Scholar
  48. Wilson, T., Hewlett, S.J., and Sheppard, C.J.R., 1990, Use of objective lenses with slit pupil functions in the imaging of line structures, Appl. Optics 29:4705–4714. The gradient of the image of a straightedge is 17.8% sharper if one of the lenses has a slit pupil function.CrossRefGoogle Scholar
  49. Xiao, G.Q., Corle, T.R., and Kino, G.S., 1988, Real-time confocal scanning optical microscope, Appl. Phys Lett. 53:716–718. This describes the one-sided TSM from Kino’s lab—Now available from Technical Instruments.CrossRefGoogle Scholar
  50. Young, M.R., Jiang, S.H., Davies, R.E., Walker, J.G., Pike, E.R., and Bertero, M., 1992, Experimental confirmation of super-resolution in coherent confocal scanning microscopy using optical masks, J. Microsc. 165:131–138. Super-resolution in a coherent scanning microscope with a special optical mask, a Fourier lens, and detector pinhole to carry out optical processing of the image. The form of the special mask was calculated using the theory of singular systems.CrossRefGoogle Scholar


  1. Arndt-Jovin, D.J., Robert Nicoud, M., and Jovin, T.M., 1990, Probing DNA structure and function with a multi-wavelength fluorescence confocal laser microscope, J. Microsc. 157:61–72 Three levels of organization in DNA structure in the interphase cell nucleus are assessed by CSM..PubMedCrossRefGoogle Scholar
  2. Becker, D.L., Dekkers, J., Navarrete, R., Green, C.R., and Cook, J.E., 1991, Enhancing the laser scanning confocal microscopic visualization of Lucifer Yellow filled cells in whole-mounted tissue, Scanning Microsc. 5:619–624. Clearing Lucifer Yellow-injected neurons in methyl salicylate and mounting them in Entellan enhances their visualization with fluorescein and reduces photobleaching.PubMedGoogle Scholar
  3. Carlsson, K., Wallen, P., and Brodin, L., 1989, Three-dimensional imaging of neurons by confocal fluorescence microscopy, J. Microsc. 155:15–26. Specimen preparation, the instrument and its performance are described. The limits in photometric quality are set by photon quantum noise.PubMedCrossRefGoogle Scholar
  4. Cohen, A.R., Roysam, B., and Turner, J.N., 1994, Automated tracing and volume measurements of neurons from 3-D confocal fluorescence microscopy data, J. Microsc. 173:103–114. A recent example of the extraction of measurements from CSM data.PubMedCrossRefGoogle Scholar
  5. Dirnagl, U., Villringer, A., and Einhaupl, K.M., 1992, In vivo confocal scanning laser microscopy of the cerebral microcirculation, J. Microsc. 165:147–157. CSM to study the microcirculation of the brain neocortex in anaesthetized rats injected with fluorescein, through a cranial window, to a depth of 250μm.PubMedCrossRefGoogle Scholar
  6. Hernandez Cruz, A., Sala, F., and Adams, P.R., 1990, Subcellular calcium transients visualized by confocal microscopy in a voltage-clamped vertebrate neuron, Science 247:858–862. CSM with long-wavelength Ca2+ indicators.PubMedCrossRefGoogle Scholar
  7. Lloyd, C.W., Venverloo, C.J., Goodbody, K.C., and Shaw, P.J., 1992, Confocal laser microscopy and three-dimensional reconstruction of nucleus-associated microtubules in the division plane of vacuolated plant cells, J. Microsc. 166:99–109. Appearance and gradual reorganization of nucleus-associated microtubules (NAMTs) over the premitotic period. Epidermal explants fluorescently labeled with antitubulin were optically sectioned.CrossRefGoogle Scholar
  8. Masters, B.R., 1992, Confocal microscopy of the in situ crystalline lens, J. Microsc. 165:159–167. Images through the full thickness of the cornea and aqueous humor to a depth of 50μm in the anterior ocular lens of an excised rabbit eye.PubMedCrossRefGoogle Scholar
  9. Masters, B.R., Kriete, A., and Kukulies, J., 1993, Ultraviolet confocal fluorescence microscopy of the in vitro cornea: Redox metabolic imaging Appl. Optics 34:592–596. Active metabolites exhibit a fluorescence shift.CrossRefGoogle Scholar
  10. Mattfeldt, T., Clarke, A., and Archenhold, G., 1994, Estimation of the directional distribution of spatial fibre processes using stereology and confocal scanning laser microscopy, J. Microsc. 173:87–101. Another demonstration that CSM yields numerical data.CrossRefGoogle Scholar
  11. Montag, M., Kukulies, J., Jorgens, R., Gundlach, H., Trendelenburg, M.F., and Spring, H., 1991, Working with the confocal scanning UV-laser microscope: Specific DNA localization at high sensitivity and multiple-parameter fluorescence, J. Microsc. 163:201–210. Multiple-parameter studies and identification of the double-stranded DNA of lampbrush chromosome loops in germinal vesicles of amphibian oocytes.PubMedCrossRefGoogle Scholar
  12. Mossberg, K., and Ericsson, M., 1990, Detection of doubly stained fluorescence specimens using confocal microscopy, J. Microsc. 158:215–224. Lucifer Yellow, Texas Red, fluorescein isothiocyanate, and tetramethylrho-damine isothiocyanate.PubMedCrossRefGoogle Scholar
  13. Oud, J.L., Mans, A., Brakenhoff, G.J., van der Voort, H.T.M., van Spronsen, E.A., and Nanninga, N., 1989, Three-dimensional chromosome arrangement of crepis-capillaris in mitotic prophase and anaphase as studied by confocal scanning laser microscopy, J. Cell Sci. 92:329–340. Actual use is made of the 3D displays we see so often.PubMedGoogle Scholar
  14. Rawlins, D.J., and Shaw, P.J., 1990, Localization of ribosomal and telomeric DNA sequences in intact plant nuclei by in-situ hybridization and three-dimensional optical microscopy, J. Microsc. 157:83–89. Sections of root tips of Pisum sativum, using cDNA probes, telomeres are arranged around the nuclear periphery and ribosomal genes exist in discrete, 3D domains.PubMedCrossRefGoogle Scholar
  15. Smith, P.J., and Sykes, H.R., 1992, Simultaneous measurement of cell cycle phase position and ionizing radiation-induced DNA strand breakage in single human tumour cells using laser scanning confocal imaging, Int. J. Radial Biol. 61:553–560. Measurement of nucleoid relaxation in response to DNA damage. The volumes of spherical nucleoids and their relative DNA contents from ethidium bromide staining and equatorial sections of the nucleoids.CrossRefGoogle Scholar
  16. Valkenburg, J.A., Woldringh, C.L., Brakenhoff, G.J., van der Voort, H.T., and Nanninga, N., 1985, Confocal scanning light microscopy of the Escheri-cia coli nucleoid: Comparison with phase-contrast and electron microscope images, J. Bacteriol. 161:478–483. A comparison of E. coli strains with a phase-contrast microscope, a CSM, and an electron microscope.PubMedGoogle Scholar
  17. Van Dekken, H., Van Rotterdam, A., Jonker, R., van der Voort, H.T.M., Brakenhoff, G.J., and Bauman, J.G.J., 1990, Confocal microscopy as a tool for the study of the intranuclear topography of chromosomes, J. Microsc. 158:207–214. Centromeric regions within blood cell nuclei fluorescently labeled by in situ hybridization to suspended nuclei with a centromere-1-specific DNA probe.PubMedCrossRefGoogle Scholar
  18. van Meer, G., Stelzer, E.H.K., Wijnaendts van Resandt, R.W., and Simons, K., 1987, Sorting of sphingolipids in epithelial (Madin-Darby canine kidney) cells, J. Cell Biol. 105:1623–1635. The fluorescent marker most likely concentrated in the Golgi complex itself.PubMedCrossRefGoogle Scholar
  19. Wikler, K.C., and Rakic, P., 1991, Relation of an array of early-differentiating cones to the photoreceptor mosaic in the primate retina, Nature 351:397 – 400. The inner segments of immunolabeled and surrounding unlabeled cones are transiently in apposition with one another.PubMedCrossRefGoogle Scholar
  20. Xiao, G.Q., Kino, G.S., and Masters, B.R., 1990, Observation of the rabbit cornea and lens with a new real-time confocal scanning optical microscope, Scanning 12:161–166. Excised rabbit eye.CrossRefGoogle Scholar

Variants on the Main Theme

  1. Bailey, B., Farkas, D.L., Taylor, D.L., and Lanni, F., 1993, Enhancement of axial resolution in fluorescence microscopy by standing-wave excitation, Nature 366:44–48. A means of increasing axial sectioning resolution at the expense of an uncertainty in choice of which standing wave plane is addressed. Combined with CSM, an improvement to 0.05μm is claimed, although the standing waves are λ/2ŋ apart (0.15 μm).PubMedCrossRefGoogle Scholar
  2. Benschop, J., and van Rosmalen, G., 1991, Confocal compact scanning optical microscope based on compact disc technology, Appl. Optics 30:1179–1184. A compact CSM based on the optics and mechanics of a compact disc (CD) player is equipped with automatic focusing. The laser in the CD player is replaced by the endface of a single-mode fiber which acts both as the emitting source and as a point detector.CrossRefGoogle Scholar
  3. Burns, D.H., Hatanagadi, R.B., and Spelman, F.A., 1990, Scanning slit aperture confocal microscopy for three-dimensional imaging, Scanning 12:156 – 160. A CSM using stationary slit apertures of variable width employing an oscillating mirror at 150 frames/sec.CrossRefGoogle Scholar
  4. Carter, K.C., Bowman, D., Carrington, W., Fogarty, K., McNeil, J.A., Fay, F.S., and Lawrence, J.B., 1993, A three-dimensional view of precursor messenger RNA metabolism within the mammalian nucleus, Science 259:1330–1335. A demonstration of the alternative (software) approach of Carrington and Fay, in real use.PubMedCrossRefGoogle Scholar
  5. Chim, S.S.C., and Kino, G.S., 1990, Correlation microscope, Opt. Lett. 15:579–581. A Mirau interferometer used with Kino’s disk-scanning confocal microscope allows collection of both amplitude and phase information.PubMedCrossRefGoogle Scholar
  6. Ching Bo Juang, Finzi, L., and Bustamante, C.J., 1988, Design and application of a computer-controlled confocal scanning differential polarization microscope, Rev. Sci. Instrum. 59:2399–2408. Basic theory and design. Anisotropy 10~5, typical CSM resolutionsCrossRefGoogle Scholar
  7. Cohen-Sabban, J., Rodier, J.C., Roussel, A., and Simon, J., 1984, Scanning ophthalmoscope, Innov. Tech. Biol. Med. 5:24 A scanning laser ophthalmoscope with two galvo mirrors..Google Scholar
  8. Curley, P.F., Ferguson, A.I., White, J.G., and Amos, W.B., 1992, Application of a femtosecond self-sustaining mode-locked Ti:sapphire laser to the field of laser scanning confocal microscopy, Opt. Quantum Electron. 24:851–859. With a femtosecond Ti:sapphire laser in a CSM, spectra are given for various fluorescent stains under two-photon excitation.CrossRefGoogle Scholar
  9. Denk W., Strickler J.H., and Webb W.W., 1990, Two-photon laser scanning fluorescence in microscopy, Science 248:73–76. Two infrared photons excite the same fluorescence as one ultraviolet photon, but there must be a lot of IR in one small volume. No confocal pinhole is necessary, since the requisite convolution of two J 1(r)2/r 2 functions comes from the two illuminations.PubMedCrossRefGoogle Scholar
  10. Draaijer, A., and Houpt, P.M., 1988, A standard video-rate confocal laser-scanning reflection and fluorescence microscope, Scanning 10:139–145. The CSM developed at TNO has standard video-rate imaging, and is capable of working in reflection and in fluorescence mode simultaneously.CrossRefGoogle Scholar
  11. Gimitri, A.F., and Aziz, D., 1993, Confocal microscopy through a fiber-optic imaging bundle, Opt. Lett. 18:565–567. A coherent fiber bundle relays the scan between objectives, presently with low-power objectives.CrossRefGoogle Scholar
  12. Goldstein, S.R., Hubin, T., Rosenthal, S., and Washburn, C., 1990, A confocal video-rate laser-beam scanning reflected-light microscope with no moving parts, J. Microsc. 157:29–38. Acousto-optic scanners for the incident beam, and confocal descanning and detection by an image dissector tube.PubMedCrossRefGoogle Scholar
  13. Hell, S., and Stelzer, E.H.K., 1992, Fundamental improvement of resolution with a 4Pi-confocal fluorescence microscope using two-photon excitation, Opt. Commun. 93:277–282. An implementation of Watt Webb’s microscope is proposed. Calculations demonstrate that an axial resolution on the order of 100 nm can be achieved in a confocal microscope with enlarged aperture when two-photon excitation is applied. The proposed microscope yields the highest point resolution ever achieved in far-field microscopy.CrossRefGoogle Scholar
  14. Hiraoka, Y., Minden, J.S., Swedlow, J.R., Sedat, J.W., and Agard, D.A., 1989, Focal points for chromosome condensation and decondensation from three-dimensional in vivo time-lapse microscopy, Nature 342:293–296. Chromosomes can now be studied in the transitions to and from interphase. Demonstrates 3D analysis, time lapse display, and the (software) emulation of confocal microscopy.PubMedCrossRefGoogle Scholar
  15. Jungerman, R.L., Hobbs, P.C., and Kino, G.S., 1984, Phase sensitive scanning optical microscope, Appl. Phys. Lett. 45:846–848. An electronically scanned optical microscope measuring amplitude and phase and insensitive to mechanical vibrations. Phase information gives an axial accuracy of 10 nm and can improve the lateral resolution.CrossRefGoogle Scholar
  16. Juskaitis, R., Wilson, T., and Reinholtz, F., 1993, Spatial filtering by laser detection in confocal microscopy, Opt. Lett. 18:1135–1137. The laser itself is the confocal pinhole, and is modulated by the remitted light.PubMedCrossRefGoogle Scholar
  17. Koester, C.J., 1980, Scanning mirror microscope with optical sectioning characteristics: Applications in ophthalmology, Appl. Optics 19:1749–1757. The original paper on Koester’s real-time slit scanner.CrossRefGoogle Scholar
  18. Koester, C.J., Auran, J.D., Rosskothen, H.D., Florakis, G.J., and Tackaberry, R.B., 1993, Clinical microscopy of the cornea utilizing optical sectioning and a high-numerical-aperture objective, J. Opt. Soc. Am. A 10:1670–1679. Use of the specular microscope with a dipping cone and film recording.PubMedCrossRefGoogle Scholar
  19. Lichtman, J.W., Sunderland, W.J., and Wilkinson, R.S., 1989, High-resolution imaging of synaptic structure with a simple confocal microscope, The New Biologist 1:75–82. An oscillating slit at a relayed image plane turns a conventional microscope into a TSM variant. Origin of the Newport CSM.PubMedGoogle Scholar
  20. McLaren, J.W., and Brubaker, R.F., 1988, A scanning ocular spectrofluoro-photometer, Invest. Ophthalmol. Vis. Sci. 29:1285–1293. Measures fluorescence in a 2D cross section through the anterior chamber and cornea of the eye.PubMedGoogle Scholar
  21. Morgan, C.G., Mitchell, A.C., and Murray, J.G., 1992, Prospects for confocal imaging based on nanosecond fluorescence decay time, J. Microsc. 165:49–60. Two detection schemes: one based on an imaging single photon detector equipped with an external photon correlator and the other using an rf-modulated intensified CCD camera. Possible routes to development of a confocal version of the microscope.CrossRefGoogle Scholar
  22. Piston, D.W., Kirby, M.S., Cheng, H., Lederer, W.J., and Webb, W.W., 1994, Two-photon-excitation fluorescence imaging of three-dimensional calcium-ion activity, Appl. Optics 33:662. Actual imaging with the two-photon microscope. Some 50 mW of near IR in 100-fs pulses into an MRC 600.CrossRefGoogle Scholar
  23. Puppels, G.J., Colier, W., Olminkhof, J.H.F., Otto, C., de Mul, F.F.M., and Grève, J., 1991, Description and performance of a highly sensitive confocal Raman microspectrometer, J. Raman Spectrosc. 22:217–225. Lateral resolution 0.45μm and 1.3μm axial, for single living cells and metaphase and polytene chromosomes. Excitation 660 nm to avoid sample degradation. 1000 cm-1 Raman shift. For signal detection a liquid nitrogen-cooled slow-scan CCD camera is used. Laser powers of 5–10 mW suffice to obtain high-quality Raman spectra, with signal integration times of the order of minutes. As an example, spectra obtained from the nucleus and the cytoplasm of an intact human lymphocyte are shown.CrossRefGoogle Scholar
  24. Sawatari, T., 1973, Optical heterodyne scanning microscope, Appl. Optics 12:2768–2772. Image similar to CSM because the same self convolution of the diffraction pattern is required. Ambient light effects eliminated. High contrast images from low-contrast objects.CrossRefGoogle Scholar
  25. van Norren, D., and van de Kraats, J., 1989, Imaging retinal densitometry with a confocal scanning laser ophthalmoscope, Vis. Res. 29:1825–1830. Use of the scanning laser ophthalmoscope (SLO) as an imaging retinal densitometer.PubMedCrossRefGoogle Scholar
  26. Webb, R.H., 1984, Optics for laser rasters, Appl. Optics 23:3680–3683. Optics for shaping and manipulating laser beams in a video scanning system, including an awkward but useable solution to the problem of chromaticity in diffraction scanners.CrossRefGoogle Scholar
  27. Webb, R.H., Hughes, G.W., and Delori, F.C., 1987, Confocal scanning laser ophthalmoscope, Appl. Optics 26:1492–1499. The SLO sold by Roden-stock. Works in real time, using an avalanche photodiode as detector. Last in a series on this instrument.CrossRefGoogle Scholar

Fiber Optic CSMs

  1. Dabbs, T., and Glass, M., 1992, Single-mode fibers used as confocal microscope pinholes, Appl. Optics 31:705–706, and Dabbs, T., and Glass, M., 1992, Fiber-optic confocal microscope: FOCON, Appl. Optics 31:3030–3035. The beamsplitter is replaced by a fiber-optic splitter, and the core of a single-mode fiber takes the place of both the source and detector pinholes. Scanning in the x, y, and z directions is by moving the end of the optical fiber.CrossRefGoogle Scholar
  2. Gan, X., Gu, M., and Sheppard, C.J.R., 1992, Fluorescent image formation in the fibre-optical confocal scanning microscope, J. Mod. Opt. 39:825–834. Unlike a finite circular pinhole, there is no missing cone of spatial frequencies, and no negative tail in the transfer function.CrossRefGoogle Scholar
  3. Ghiggino, K.P., Harris, M.R., and Spizzirri, P.G., 1992, Fluorescence lifetime measurements using a novel fibre-optic laser scanning confocal microscope, Rev. Sci. Instrum. 63:2999–3002. A fiber-optic CSM with a mode-locked dye laser excitation source, avalanche photodiode detector, and time-correlated photon counting electronics, allows spatially resolved fluorescence decay profiles from fluorescent dyes in solution and polymer films.CrossRefGoogle Scholar
  4. Giniunas, L., Juskaitis, R., and Shatalin, S.V., 1991 Scanning fibre-optic microscope, Electron. Lett. 27:724–726. A phase-sensitive scanning fiber-optic microscope designed to be used as an endoscope. Confocal operation with single mode fiber yields 0.8 μm lateral resolution.CrossRefGoogle Scholar
  5. Gu, M., and Sheppard, C.J.R., 1991, Signal level of the fibre-optical CSM, J. Mod. Opt. 38:1621–1630. The efficiency of total power transformation and signal level of the fiber-optical CSM, including defocus effects.CrossRefGoogle Scholar
  6. Gu, M., and Sheppard, C.J.R., 1992, Axial resolution in the fibre-optical confocal scanning microscope using annular lenses, Opt. Commun. 88:27–32, and Gu, M., and Sheppard, C.J.R., 1992, Image of a straight edge in fibre-optical confocal scanning microscopy, Opt. Commun. 94:455–490. Relationship of the central obstruction to the fiber spot size.CrossRefGoogle Scholar
  7. Gu, M., Sheppard, C.J.R., and Gan, X., 1991, Image formation in a fiber-optical confocal scanning microscope, J. Opt. Soc. Am. A, Opt. Image Sci. 8:1755–1761. Coherent transfer functions in both in-focus and defocused cases are derived and calculated.CrossRefGoogle Scholar
  8. Juskaitis, R., and Wilson, T., 1992, Differential CSM with a two-mode optical fiber, Appl. Optics 31:898–902. Both the differential amplitude and the differential phase images can be obtained by adjusting the differential phase delay between the fiber modes.CrossRefGoogle Scholar
  9. Juskaitis, R., and Wilson, T., 1992, Imaging in reciprocal fibre-optic based CSMs, Opt. Commun. 92:315–325. Single- and two-mode fibers in a reciprocal scheme. Confocal, differential amplitude or phase contrast imaging and surface profilometry with reduced alignment tolerances.CrossRefGoogle Scholar
  10. Juskaitis, R., Reinholz, F., and Wilson, T., 1992, Fibre-optic based confocal scanning microscopy with semiconductor laser excitation and detection, Electron. Lett. 28:986–988. Collected light reenters the laser and the image is detected as a modulation of the laser power.CrossRefGoogle Scholar
  11. Kimura, S., and Wilson, T., 1991, Confocal scanning optical microscope using single-mode fiber for signal detection, Appl. Optics 30:2143–2150. Always coherent, which is fundamentally different from a finite-sized pinhole.CrossRefGoogle Scholar


  1. Corle, T.R., Fanton, J.T., and Kino, G.S., 1987, Distance measurements by differential confocal optical ranging, Appl. Optics 26:2416–2420. Dithering generates a differential measurement, placing a zero-crossing at the peak of the depth response. Sensitivities to surface vibrations of 0.01 nm and thin film measurements to 0.04 μmCrossRefGoogle Scholar
  2. Hamilton, D.K., and Matthews, H.J., 1985, The confocal interference microscope as a surface profilometer, Optik 71:31–34. A noncontacting high-resolution surface profiling technique, with feedback to maintain a constant phase relationship between the signal and the reference beams of a confocal interference microscope.Google Scholar
  3. Hamilton, D.K., and Wilson, T., 1982, Three-dimensional surface measurement using the confocal scanning microscope, Appl. Phys. B 27:211–213. 0.1 urn axial resolution on a semiconductor.CrossRefGoogle Scholar
  4. Lee, B.S., and Strand, T.C., 1990, Profilometry with a coherence scanning microscope, Appl. Optics 29:3784–3788. Uses coherence effects rather than physical apertures. Longitudinal resolution is decoupled from lateral.CrossRefGoogle Scholar
  5. Matthews, H.J., Hamilton, D.K., and Sheppard, C.J.R., 1986, Surface profiling by phase-locked interferometry, Appl. Optics 25:2372–2374. 1-nm height resolution. A feedback arrangement keeps in quadrature the two arms of a confocal interference microscope by modulating the reference beam using an electro-optic phase modulator.CrossRefGoogle Scholar
  6. Toy, D.A., 1990, Confocal microscopy: The ups and downs of 3-D profiling. Semicond. Int. 13:120–123. Confocal techniques can solve the problems of depth of field versus resolution.Google Scholar


  1. Carlsson, K., Danielsson, P.E., Lenz, R., Liljeborg, A., Majlof, L., and Åslund, N., 1985, Three-dimensional microscopy using a confocal laser scanning microscope, Opt. Lett. 10:53–55. Visualizes in stereo and rotation by making look-through projections of the digital data in different directions. Contrast enhanced by generating gradient volume to display the border surfaces between regions in opposing senses for the two images.PubMedCrossRefGoogle Scholar
  2. Conchello, J.A., and Hansen, E.W., 1990, Enhanced 3-D reconstruction from CSM images. I. Deterministic and maximum likelihood reconstructions, Appl. Optics 29:3795–3804. The feasibility of obtaining longitudinal resolution comparable to the lateral diffraction limit by posterior processing of confocal sections.CrossRefGoogle Scholar
  3. Dilworth, D.S., Leith, E.N., and Lopez, J.L., 1991, Three-dimensional confocal imaging of objects embedded within thick diffusing media, Appl. Optics 30:1796–1803. Exfoliative deconvolution to sharpen a volume image in which the blur is depth variant.CrossRefGoogle Scholar
  4. Hiraoka, Y., Sedat, J.W., and Agard, D.A., 1990, Determination of three-dimensional imaging properties of a light microscope system. Partial confocal behavior in epifluorescence microscopy, Biophys. J. 57:325–333. Through-focus series of a point object were recorded on a charge-coupled device image detector. From these images, the 3D point spread function and its Fourier transform, the optical transfer function, were derived for use in processing a CSM image.PubMedCrossRefGoogle Scholar
  5. Oldmixon, E.H., and Carlsson, K., 1993, Methods for large data volumes from confocal scanning laser microscopy of lung, J. Microsc. 170:221–228. How to stitch together adjacent small fields to achieve high resolution over a large area (500 resels per diameter). Intensity variations are adjusted for uniformity of measurement.PubMedCrossRefGoogle Scholar
  6. Schormann, T., and Jovin, T.M., 1992, Contrast enhancement and depth perception in three-dimensional representations of differential interference contrast and CSM images, J. Microsc. 166:155–168. A new contrast enhancement transformation based upon local statistics and a gray-level probability density function provides improved depth perception, increasing the number of usable optical sections by up to five.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 1995

Authors and Affiliations

  • Robert H. Webb
    • 1
  1. 1.Wellman LaboratoriesSchepens Eye Research Institute and MGH Laser CenterBostonUSA

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