OSA's Digital Library

Applied Optics

Applied Optics


  • Editor: Joseph N. Mait
  • Vol. 50, Iss. 28 — Oct. 1, 2011
  • pp: 5524–5533

New method for determining the depth of field of microscope systems

Xiaodong Chen, Liqiang Ren, Yuchen Qiu, and Hong Liu  »View Author Affiliations

Applied Optics, Vol. 50, Issue 28, pp. 5524-5533 (2011)

View Full Text Article

Enhanced HTML    Acrobat PDF (720 KB)

Browse Journals / Lookup Meetings

Browse by Journal and Year


Lookup Conference Papers

Close Browse Journals / Lookup Meetings

Article Tools



This paper presents new formulas to determine the depth of field (DOF) of optical and digital microscope systems. Unlike the conventional DOF formula, the new methods consider the interplay of geometric and diffraction optics for infinite and finite optical microscopes and for corresponding digital microscope systems. It is shown that in addition to the well understood parameters such as numerical apertures, focal length, and light wavelength, system components such as aperture stops also affect the DOF. For the same objective lens, the DOF is inversely proportional to the size of the aperture stop, and it is proportional to the focal length of the ocular lens. It is also shown that under optimal viewing and operating conditions, the visual accommodation of human observers has no meaningful impact on DOF. The new formulas reported are useful for accurately calculating the DOF of microscopes.

© 2011 Optical Society of America

OCIS Codes
(110.0110) Imaging systems : Imaging systems
(110.0180) Imaging systems : Microscopy
(260.0260) Physical optics : Physical optics
(260.1960) Physical optics : Diffraction theory

Original Manuscript: April 20, 2011
Revised Manuscript: July 3, 2011
Manuscript Accepted: August 8, 2011
Published: September 30, 2011

Virtual Issues
Vol. 6, Iss. 11 Virtual Journal for Biomedical Optics

Xiaodong Chen, Liqiang Ren, Yuchen Qiu, and Hong Liu, "New method for determining the depth of field of microscope systems," Appl. Opt. 50, 5524-5533 (2011)

Sort:  Author  |  Year  |  Journal  |  Reset  


  1. P. Gualtieri and L. Barsanti, “Identification of cellular and subcellular features by means of digital microscopy,” Int. J. Biomed. Comput. 20, 79–86 (1987). [CrossRef] [PubMed]
  2. J. W. Bacus and L. J. Grace, “Optical microscope system for standardized cell measurements and analysis,” Appl. Opt. 26, 3280–3293 (1987). [CrossRef] [PubMed]
  3. I. J. Cox and J. R. Sheppard, “Scanning optical microscope incorporating a digital framestore and microcomputer,” Appl. Opt. 22, 1474–1478 (1983). [CrossRef] [PubMed]
  4. M. S. Elliot and W. C. K. Poon, “Conventional optical microscopy of colloidal suspensions,” Adv. Colloid Interface Sci. 92, 133–194 (2001). [CrossRef] [PubMed]
  5. S. Yasuda, D. N. Futaba, M. Yumura, S. Iijima, and K. Hata, “Diagnostics and growth control of single-walled carbon nanotube for using a telecentric optical system for in situ height monitoring,” Appl. Phys. Lett. 93, 143115 (2008). [CrossRef]
  6. C. Maurer, S. Khan, S. Fassl, S. Bernet, and M. Ritsch-Marte, “Depth of field multiplexing in microscopy,” Opt. Express 18, 3023–3034 (2010). [CrossRef] [PubMed]
  7. S. Liu and H. Hua, “Extended depth-of-field microscopic imaging with a variable focus microscope objective,” Opt. Express 19, 353–362 (2011). [CrossRef] [PubMed]
  8. H. Piller, Microscope Photometry, Vol. 16 (Springer-Verlag, 1977), p. 16.
  9. L. C. Martin, The Theory of the Microscope (Elsevier, 1966), pp. 192–204.
  10. M. Born and E. Wolf, with contributions by A. B. Bhatia, P. C. Clemmow, D. Gabor, A. R. Stokes, A. M. Taylor, P. A. Wayman, and W. L. Wilcock, Principles of Optics: Electromagnetic Theory of Propagation, Interference and Diffraction of Light, 7th (expanded) ed. (Cambridge University, 1999). [PubMed]
  11. B. Michael, Handbook of Optics Volume I: Geometrical and Physical Optics, Polarized Light, Components and Instruments, 3rd ed. (McGraw-Hill, 2010). [PubMed]
  12. W. J. Smith, Modern Optical Engineering, 4th ed. (McGraw-Hill, 2008).
  13. S. Inoue and K. R. Spring, Video Microscopy: the Fundamentals (Plenum, 1997). [CrossRef]
  14. H. Gross, H. Zügge, M. Peschka, and F. Blechinger, Handbook of Optical Systems, Vol.  3 (Wiley-VCH, 2007), p. 125.
  15. D. Hong, K. Park, H. Cho, and M. Kim, “Flexible depth-of-field imaging system using a spatial light modulator,” Appl. Opt. 46, 8591–8599 (2007). [CrossRef] [PubMed]
  16. G. Häusler, “A method to increase the depth of focus by two step image processing,” Opt. Commun. 6, 38–42 (1972). [CrossRef]
  17. P. Green, W. Sun, W. Matusik, and F. Durand, “Multi-aperture photography,” ACM Trans. Graph. 26, 68 (2007). [CrossRef]
  18. A. Levin, S. W. Hasinoff, P. Green, F. Durand, and W. T. Freeman, “4D frequency analysis of computational cameras for depth of field extension,” ACM Trans. Graph. 28, 97 (2009). [CrossRef]
  19. F. Diaz, F. Goudail, B. Loiseaux, and J.-P. Huignard, “Increase in depth of field taking into account deconvolution by optimization of pupil mask,” Opt. Lett. 34, 2970–2972(2009). [CrossRef] [PubMed]
  20. B. Chebbi, S. Minko, N. Al-Akwaa, and I. Golub, “Remote control of extended depth of field focusing,” Opt. Commun. 283, 1678–1683 (2010). [CrossRef]

Cited By

Alert me when this paper is cited

OSA is able to provide readers links to articles that cite this paper by participating in CrossRef's Cited-By Linking service. CrossRef includes content from more than 3000 publishers and societies. In addition to listing OSA journal articles that cite this paper, citing articles from other participating publishers will also be listed.

« Previous Article  |  Next Article »

OSA is a member of CrossRef.

CrossCheck Deposited