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Applied Optics

Applied Optics

APPLICATIONS-CENTERED RESEARCH IN OPTICS

  • Editor: Joseph N. Mait
  • Vol. 49, Iss. 15 — May. 20, 2010
  • pp: 2954–2968

Sample-less calibration of the differential interference contrast microscope

Shalin B. Mehta and Colin J. R. Sheppard  »View Author Affiliations


Applied Optics, Vol. 49, Issue 15, pp. 2954-2968 (2010)
http://dx.doi.org/10.1364/AO.49.002954


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Abstract

Analysis of image formation in a differential interference contrast (DIC) microscope and retrieval of the specimen’s properties require calibration of its key parameters, viz. shear and bias. We present a method of measuring the shear and the bias of a DIC microscope from the interference fringes produced in the back focal plane of the objective. Previous approaches, which use calibrated specimens such as polysty rene or fluorescent beads, provide rather approximate measurements of shear or require a complex optical setup. The method presented is accurate, relies on simple image analysis, and does not require a specimen. We provide a succinct and accurate description of properties of Nomarski prisms to explain the rationale behind the method.

© 2010 Optical Society of America

OCIS Codes
(110.2960) Imaging systems : Image analysis
(110.2990) Imaging systems : Image formation theory
(110.4980) Imaging systems : Partial coherence in imaging
(180.3170) Microscopy : Interference microscopy

ToC Category:
Microscopy

History
Original Manuscript: January 11, 2010
Revised Manuscript: April 8, 2010
Manuscript Accepted: April 9, 2010
Published: May 19, 2010

Virtual Issues
Vol. 5, Iss. 10 Virtual Journal for Biomedical Optics

Citation
Shalin B. Mehta and Colin J. R. Sheppard, "Sample-less calibration of the differential interference contrast microscope," Appl. Opt. 49, 2954-2968 (2010)
http://www.opticsinfobase.org/ao/abstract.cfm?URI=ao-49-15-2954

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Supplementary Material


» Media 1: MOV (3749 KB)     
» Media 2: MOV (1427 KB)     
» Media 3: MOV (748 KB)     
» Media 4: MOV (1204 KB)     
» Media 5: MOV (1357 KB)     

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