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Diffracted Light Contrast: Improving the Resolution of a Basic Light Microscope by an Order of Magnitude

Published online by Cambridge University Press:  14 March 2018

W. Barry Piekos
W. Barry Piekos*
Affiliation:
Yale University, New Haven, CT
*
[email protected]

Extract

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The discovery that the diffracted light from a convex edge can be used to form a very high-quality, shadowcast image on any light microscope has led to a device and method, diffracted-light contrast (DLC), which will allow shadowcast imaging to be routinely performed on student/laboratory microscopes (Piekos, 1999, 2003). The surface lattice of Surirella gema was easily resolved, and micrographs comparing the subcellular details of buccal epithelial cells viewed with DLC vs. Nomarski DIC showed that, on the microscopes used, DLC was superior in both the detail it rendered and depth of field. Although the images presented revealed DLC to be an excellent technique, the full capabilities of the technique were not known at the time.

Type
Research Article
Information
Microscopy Today , Volume 14 , Issue 6 , November 2006 , pp. 10 - 15
Copyright
Copyright © Microscopy Society of America 2006

References

Axelrod, D. (1980) Zero-cost modification of bright field microscopes for imaging phase gradient on cells: schlieren optics. Cell Biophys 3, 167–173.Google Scholar
Cogswell, C.J. (1995) In Handbook of Biological Confocal Microscopy, Second Edition (ed. Pawley, J.B.). Plenum Press, New York and London, pp. 507–513.Google Scholar
de Lange, E, Cambi, A., Huijbens, R., de Bakker, B., Rensen, W., Garcia-Parajo, M., van Hulst, N. & Figdor, C.G. (2001) Cell biology beyond the diffraction limit: near-field scanning optical microscopy. / Cell Sci, 114,4153-4 160.Google Scholar
Diggins, B.A. (1940) Oblique light diaphragm. U.S. Patent 2,195,166.Google Scholar
Gonzalez-Melendi, P., Boudonek, K., Abranches, R., Wells, B., Dolan, L. & Shaw, P. (2000) The nucleus: a highly organized but dynamic structure. / Microscopy 198(3), 199–207.Google Scholar
Gustafsson, M.G.L., Agard, D.A. & Sedat, J.W. (1999) I5M: 3D widefield light microscopy with better than 100 nm axial resolution. /. Microscopy 195(1), 10–16.CrossRefGoogle ScholarPubMed
Ott, H.N. (1924) Oblique light diaphragm. U.S. Patent 1,503,800.Google Scholar
Piekos, W.B. (1999) Diffracted-light contrast enhancement: a re-examination of oblique illumination. Micros Res Tech 46, 334–337.Google Scholar
Piekos, W.B. (2003) Apparatus and method for producing diffracted light contrast enhancement in microscopes. U.S. Patent 6,600,598.Google Scholar
Saylor, C.P. (1935) Accuracy of microscopical methods for determining refractive index by immersion. / Res Nat Bur Standards 15, 277–294.Google Scholar
Schrader, M., Hell, S.W & van der Voort, H.T.M. (1998) Three-dimensional superresolution with a 4Pi-confocal microscope using image restoration. / Applied Physics 84(8), 4033–4042.Google Scholar
Watanabe, T, Iketaki, Y., Omatsu, T., Yamamoto, K., Sakai, M. & Fujii, M. (2003) Two-point-separation in super-resolution fluorescence microscope based on up-conversion fluorescence depletion technique. Optics Express 11(24), 3271–3276.Google Scholar
Wright, F.E. (1913) Oblique illumination in petrographic microscope work. Am J Sci 205, 63–82.CrossRefGoogle Scholar