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Biomedical Optics Express

Biomedical Optics Express

  • Editor: Joseph A. Izatt
  • Vol. 5, Iss. 5 — May. 1, 2014
  • pp: 1588–1609

Optimal lens design and use in laser-scanning microscopy

Adrian Negrean and Huibert D. Mansvelder  »View Author Affiliations

Biomedical Optics Express, Vol. 5, Issue 5, pp. 1588-1609 (2014)

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In laser-scanning microscopy often an off-the-shelf achromatic doublet is used as a scan lens which can reduce the available diffraction-limited field-of-view (FOV) by a factor of 3 and introduce chromatic aberrations that are scan angle dependent. Here we present several simple lens designs of superior quality that fully make use of high-NA low-magnification objectives, offering diffraction-limited imaging over a large FOV and wavelength range. We constructed a two-photon laser-scanning microscope with optimized custom lenses which had a near diffraction limit point-spread-function (PSF) with less than 3.6% variation over a 400 µm FOV and less than 0.5 µm lateral color between 750 and 1050 nm.

© 2014 Optical Society of America

OCIS Codes
(180.0180) Microscopy : Microscopy
(180.1790) Microscopy : Confocal microscopy
(220.3620) Optical design and fabrication : Lens system design
(220.3630) Optical design and fabrication : Lenses
(180.4315) Microscopy : Nonlinear microscopy

ToC Category:

Original Manuscript: January 17, 2014
Revised Manuscript: March 23, 2014
Manuscript Accepted: March 23, 2014
Published: April 18, 2014

Adrian Negrean and Huibert D. Mansvelder, "Optimal lens design and use in laser-scanning microscopy," Biomed. Opt. Express 5, 1588-1609 (2014)

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  1. W. Denk, J. H. Strickler, and W. W. Webb, “Two-photon laser scanning fluorescence microscopy,” Science248(4951), 73–76 (1990). [CrossRef] [PubMed]
  2. F. Helmchen, K. Svoboda, W. Denk, and D. W. Tank, “In vivo dendritic calcium dynamics in deep-layer cortical pyramidal neurons,” Nat. Neurosci.2(11), 989–996 (1999). [CrossRef] [PubMed]
  3. F. Helmchen and W. Denk, “Deep tissue two-photon microscopy,” Nat. Methods2(12), 932–940 (2005). [CrossRef] [PubMed]
  4. P. Theer and W. Denk, “On the fundamental imaging-depth limit in two-photon microscopy,” J. Opt. Soc. Am. A23(12), 3139–3149 (2006). [CrossRef] [PubMed]
  5. G. Cox and C. J. R. Sheppard, “Practical limits of resolution in confocal and non-linear microscopy,” Microsc. Res. Tech.63(1), 18–22 (2004). [CrossRef] [PubMed]
  6. M. Oheim, E. Beaurepaire, E. Chaigneau, J. Mertz, and S. Charpak, “Two-photon microscopy in brain tissue: parameters influencing the imaging depth,” J. Neurosci. Methods111(1), 29–37 (2001). [CrossRef] [PubMed]
  7. D. Kobat, N. G. Horton, and C. Xu, “In vivo two-photon microscopy to 1.6-mm depth in mouse cortex,” J. Biomed. Opt.16(10), 106014 (2011). [CrossRef] [PubMed]
  8. A. Nagler, “Plossl type eyepiece for use in astronomical instruments,” US Patent 4,482,217 (1984).
  9. R. E. Fischer, B. Tadic-Galeb, and P. R. Yoder, Optical System Design, 2nd ed. (The McGraw-Hill Companies, Inc., 2008), pp. 211–212.
  10. J. B. Pawley, “The intermediate optical system of laser-scanning confocal microscopes,” in Handbook of Biological Confocal Microscopy, 3rd ed. (Springer, 2006), pp. 207–208.
  11. W. J. Smith, Modern Optical Engineering, 4th ed. (McGraw-Hill Professional, 2007), pp. 457–462.
  12. Q. T. Nguyen, N. Callamaras, C. Hsieh, and I. Parker, “Construction of a two-photon microscope for video-rate Ca(2+) imaging,” Cell Calcium30(6), 383–393 (2001). [CrossRef] [PubMed]
  13. W. Denk, D. W. Piston, and W. W. Webb, “Two-photon molecular excitation in laser scanning microscopy,” in Handbook of Biological Confocal Microscopy, J. B. Pawley, ed., 2nd ed. (Plenum Press, New York, 1995), pp. 445–458.
  14. M. Seel, “Beam deflection device,” US Patent 6,433,908 (2002).
  15. G. Sharafutdinova, J. Holdsworth, and D. van Helden, “Improved field scanner incorporating parabolic optics. Part 1: simulation,” Appl. Opt.48(22), 4389–4396 (2009). [CrossRef] [PubMed]
  16. G. Sharafutdinova, J. Holdsworth, and D. van Helden, “Improved field scanner incorporating parabolic optics. Part 2: Experimental verification and potential for volume scanning,” Appl. Opt.49(29), 5517–5527 (2010). [CrossRef] [PubMed]
  17. J. Schindelin, I. Arganda-Carreras, E. Frise, V. Kaynig, M. Longair, T. Pietzsch, S. Preibisch, C. Rueden, S. Saalfeld, B. Schmid, J.-Y. Tinevez, D. J. White, V. Hartenstein, K. Eliceiri, P. Tomancak, and A. Cardona, “Fiji: an open-source platform for biological-image analysis,” Nat. Methods9(7), 676–682 (2012). [CrossRef] [PubMed]
  18. M. Leutenegger, R. Rao, R. A. Leitgeb, and T. Lasser, “Fast focus field calculations,” Opt. Express14(23), 11277–11291 (2006). [CrossRef] [PubMed]
  19. M. J. Nasse and J. C. Woehl, “Realistic modeling of the illumination point spread function in confocal scanning optical microscopy,” J. Opt. Soc. Am. A27(2), 295–302 (2010). [CrossRef] [PubMed]
  20. A. Negrean and H. D. Mansvelder, “Zemax optical models from Figs. 2-9,” https://drive.google.com/folderview?id=0B4HX51TZCOZpQWVnQkVmTjFFOEk&usp=sharing

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