Abstract

We investigated the experimental performance of an afocal scan engine employing two off-axis parabolic reflectors and it was found not to introduce astigmatism when compared to a freely propagated beam. The performance of the new afocal engine is very similar to an ideal single-mirror scan engine in terms of spot size and beam spot profile (or point spread function) and has an improved flatness of field over other two-dimensional laser scan engines. The parabolic scan engine is contrasted with a comparable spherical mirror arrangement and found to produce superior performance at the intermediate image plane when focused through a scan lens. Further modeling and experimentation point toward volume scanning applications. The significant performance improvement provided by this design, now verified experimentally, will result in superior image quality for fast scanning confocal and two-photon microscopy in particular.

© 2010 Optical Society of America

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  1. H. Gross, F. Blechinger, and B. Achtner, Handbook of Optical Systems: Survey of Optical Instruments (Wiley-VCH, 2008), Vol. 4.
  2. G. F. Marshall, Handbook of Optical and Laser Scanning, 2nd ed. (CRC Press, 2004).
    [CrossRef]
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    [CrossRef]
  4. H.M.Muncheryan, ed., Laser and Optoelectronic Engineering (Hemisphere, 1991).
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    [CrossRef]
  6. E. H. K. Stelzer, “The intermediate optical system of laser-scanning confocal microscopes,” in Handbook of Biological Confocal Microscopy, J.B.Pawley, ed., 3rd ed. (Springer, 2006), pp. 207–220.
    [CrossRef]
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    [CrossRef] [PubMed]
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    [CrossRef] [PubMed]
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    [CrossRef]
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    [CrossRef]
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    [CrossRef]
  18. G. Sharafutdinova, J. Holdsworth, and D. van Helden, “Improved field scanner incorporating parabolic optics. part 1: simulation,” Appl. Opt. 48, 4389–4396 (2009).
    [CrossRef] [PubMed]
  19. G. Sharafutdinova, J. Holdsworth, and D. van Helden, “Calculated two-photon fluorescence correction factors for reflective scan engines,” Appl. Opt. 49, 1472–1479 (2010).
    [CrossRef] [PubMed]
  20. W. B. Amos, “Achromatic scanning system,” U.S. patent 4,997,242 (5 March 1991).
  21. Photon, Inc., “BeamScan Profiler,” (2009).

2010

2009

2005

J. M. Girkin and G. McConnell, “Advances in laser sources for confocal and multiphoton microscopy,” Microsc. Res. Tech. 67, 8–14 (2005).
[CrossRef] [PubMed]

2004

B. R. Masters and P. T. C. So, “Antecedents of two-photon excitation laser scanning microscopy,” Microsc. Res. Tech. 63, 3–11 (2004).
[CrossRef]

2003

W. B. Amos and J. G. White, “How the confocal laser scanning microscope entered biological research,” Biol. Cell 95, 335–342 (2003).
[CrossRef] [PubMed]

2001

J. Squier and M. Muller, “High resolution nonlinear microscopy: a review of sources and methods for achieving optimal imaging,” Rev. Sci. Instrum. 72, 2855–2867 (2001).
[CrossRef]

Achtner, B.

H. Gross, F. Blechinger, and B. Achtner, Handbook of Optical Systems: Survey of Optical Instruments (Wiley-VCH, 2008), Vol. 4.

Amos, W. B.

W. B. Amos and J. G. White, “How the confocal laser scanning microscope entered biological research,” Biol. Cell 95, 335–342 (2003).
[CrossRef] [PubMed]

W. B. Amos, “Achromatic scanning system,” U.S. patent 4,997,242 (5 March 1991).

Beiser, L.

L. Beiser, Unified Optical Scanning Technology (IEEE, 2003).
[CrossRef]

Blechinger, F.

H. Gross, F. Blechinger, and B. Achtner, Handbook of Optical Systems: Survey of Optical Instruments (Wiley-VCH, 2008), Vol. 4.

Centonze, V. E.

S. J. Wright, V. E. Centonze, S. A. Stricker, P. J. De Vries, S. W. Paddock, and G. Schatten, “An introduction to confocal microscopy and three-dimensional reconstruction,” in B.Matsumoto, ed., Cell Biological Applications of Confocal Microscopy (Academic, 1993), pp. 1–46.
[CrossRef]

Cox, G.

G. Cox, Optical Imaging Techniques in Cell Biology (CRC, 2007), p. 268.

De Vries, P. J.

S. J. Wright, V. E. Centonze, S. A. Stricker, P. J. De Vries, S. W. Paddock, and G. Schatten, “An introduction to confocal microscopy and three-dimensional reconstruction,” in B.Matsumoto, ed., Cell Biological Applications of Confocal Microscopy (Academic, 1993), pp. 1–46.
[CrossRef]

Denk, W.

W. Denk, J. P. Strickler, and W. W. Webb, “Two-photon laser microscopy,” U.S. patent 5,034,613 (23 July 1991).

Dickinson, M. E.

R. Wolleschensky, M. E. Dickinson, and S. E. Fraser, “Group-velocity dispersion and fiber delivery in multiphoton laser scanning microscopy,” in A.Diaspro, ed., Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances (Wiley-Liss, 2002), pp. 171–190.

Diels, J.-C.

J.-C. Diels and W. Rudolph, “Ultrashort laser pulse phenomena: fundamentals, techniques, and applications on a femtosecond time scale,” in Optics and Photonics (Academic, 1996), p. 581.

Fraser, S. E.

R. Wolleschensky, M. E. Dickinson, and S. E. Fraser, “Group-velocity dispersion and fiber delivery in multiphoton laser scanning microscopy,” in A.Diaspro, ed., Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances (Wiley-Liss, 2002), pp. 171–190.

Girkin, J. M.

J. M. Girkin and G. McConnell, “Advances in laser sources for confocal and multiphoton microscopy,” Microsc. Res. Tech. 67, 8–14 (2005).
[CrossRef] [PubMed]

J. M. Girkin, “Laser sources for nonlinear microscopy,” in Handbook of Biomedical Nonlinear Optical Microscopy, B.R.Masters and P.T. C.So, eds. (Oxford U. Press, 2008), p. 860.

Gross, H.

H. Gross, F. Blechinger, and B. Achtner, Handbook of Optical Systems: Survey of Optical Instruments (Wiley-VCH, 2008), Vol. 4.

Holdsworth, J.

Inoue, S.

S. Inoue, “Foundations of confocal scanned imaging in light microscopy,” in Handbook of Biological Confocal Microscopy, J.Pawley, ed., 3rd ed. (Springer, 2006), pp. 1–19.
[CrossRef]

Marshall, G. F.

G. F. Marshall, Handbook of Optical and Laser Scanning, 2nd ed. (CRC Press, 2004).
[CrossRef]

Masters, B. R.

B. R. Masters and P. T. C. So, “Antecedents of two-photon excitation laser scanning microscopy,” Microsc. Res. Tech. 63, 3–11 (2004).
[CrossRef]

McConnell, G.

J. M. Girkin and G. McConnell, “Advances in laser sources for confocal and multiphoton microscopy,” Microsc. Res. Tech. 67, 8–14 (2005).
[CrossRef] [PubMed]

Muller, M.

J. Squier and M. Muller, “High resolution nonlinear microscopy: a review of sources and methods for achieving optimal imaging,” Rev. Sci. Instrum. 72, 2855–2867 (2001).
[CrossRef]

Paddock, S. W.

S. J. Wright, V. E. Centonze, S. A. Stricker, P. J. De Vries, S. W. Paddock, and G. Schatten, “An introduction to confocal microscopy and three-dimensional reconstruction,” in B.Matsumoto, ed., Cell Biological Applications of Confocal Microscopy (Academic, 1993), pp. 1–46.
[CrossRef]

Rietdorf, J.

J. Rietdorf and E. H. K. Stelzer, “Special optical elements,” in Handbook of Biological Confocal Microscopy, J.B.Pawley, ed., 3rd ed. (Springer, 2006), pp. 43–58.
[CrossRef]

Rudolph, W.

J.-C. Diels and W. Rudolph, “Ultrashort laser pulse phenomena: fundamentals, techniques, and applications on a femtosecond time scale,” in Optics and Photonics (Academic, 1996), p. 581.

Schatten, G.

S. J. Wright, V. E. Centonze, S. A. Stricker, P. J. De Vries, S. W. Paddock, and G. Schatten, “An introduction to confocal microscopy and three-dimensional reconstruction,” in B.Matsumoto, ed., Cell Biological Applications of Confocal Microscopy (Academic, 1993), pp. 1–46.
[CrossRef]

Sharafutdinova, G.

So, P. T. C.

B. R. Masters and P. T. C. So, “Antecedents of two-photon excitation laser scanning microscopy,” Microsc. Res. Tech. 63, 3–11 (2004).
[CrossRef]

Squier, J.

J. Squier and M. Muller, “High resolution nonlinear microscopy: a review of sources and methods for achieving optimal imaging,” Rev. Sci. Instrum. 72, 2855–2867 (2001).
[CrossRef]

Stelzer, E. H. K.

E. H. K. Stelzer, “The intermediate optical system of laser-scanning confocal microscopes,” in Handbook of Biological Confocal Microscopy, J.B.Pawley, ed., 3rd ed. (Springer, 2006), pp. 207–220.
[CrossRef]

J. Rietdorf and E. H. K. Stelzer, “Special optical elements,” in Handbook of Biological Confocal Microscopy, J.B.Pawley, ed., 3rd ed. (Springer, 2006), pp. 43–58.
[CrossRef]

Stricker, S. A.

S. J. Wright, V. E. Centonze, S. A. Stricker, P. J. De Vries, S. W. Paddock, and G. Schatten, “An introduction to confocal microscopy and three-dimensional reconstruction,” in B.Matsumoto, ed., Cell Biological Applications of Confocal Microscopy (Academic, 1993), pp. 1–46.
[CrossRef]

Strickler, J. P.

W. Denk, J. P. Strickler, and W. W. Webb, “Two-photon laser microscopy,” U.S. patent 5,034,613 (23 July 1991).

van Helden, D.

Webb, W. W.

W. Denk, J. P. Strickler, and W. W. Webb, “Two-photon laser microscopy,” U.S. patent 5,034,613 (23 July 1991).

White, J. G.

W. B. Amos and J. G. White, “How the confocal laser scanning microscope entered biological research,” Biol. Cell 95, 335–342 (2003).
[CrossRef] [PubMed]

Wolleschensky, R.

R. Wolleschensky, M. E. Dickinson, and S. E. Fraser, “Group-velocity dispersion and fiber delivery in multiphoton laser scanning microscopy,” in A.Diaspro, ed., Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances (Wiley-Liss, 2002), pp. 171–190.

Wright, S. J.

S. J. Wright, V. E. Centonze, S. A. Stricker, P. J. De Vries, S. W. Paddock, and G. Schatten, “An introduction to confocal microscopy and three-dimensional reconstruction,” in B.Matsumoto, ed., Cell Biological Applications of Confocal Microscopy (Academic, 1993), pp. 1–46.
[CrossRef]

Appl. Opt.

Biol. Cell

W. B. Amos and J. G. White, “How the confocal laser scanning microscope entered biological research,” Biol. Cell 95, 335–342 (2003).
[CrossRef] [PubMed]

Microsc. Res. Tech.

J. M. Girkin and G. McConnell, “Advances in laser sources for confocal and multiphoton microscopy,” Microsc. Res. Tech. 67, 8–14 (2005).
[CrossRef] [PubMed]

B. R. Masters and P. T. C. So, “Antecedents of two-photon excitation laser scanning microscopy,” Microsc. Res. Tech. 63, 3–11 (2004).
[CrossRef]

Rev. Sci. Instrum.

J. Squier and M. Muller, “High resolution nonlinear microscopy: a review of sources and methods for achieving optimal imaging,” Rev. Sci. Instrum. 72, 2855–2867 (2001).
[CrossRef]

Other

J.-C. Diels and W. Rudolph, “Ultrashort laser pulse phenomena: fundamentals, techniques, and applications on a femtosecond time scale,” in Optics and Photonics (Academic, 1996), p. 581.

R. Wolleschensky, M. E. Dickinson, and S. E. Fraser, “Group-velocity dispersion and fiber delivery in multiphoton laser scanning microscopy,” in A.Diaspro, ed., Confocal and Two-Photon Microscopy: Foundations, Applications, and Advances (Wiley-Liss, 2002), pp. 171–190.

S. J. Wright, V. E. Centonze, S. A. Stricker, P. J. De Vries, S. W. Paddock, and G. Schatten, “An introduction to confocal microscopy and three-dimensional reconstruction,” in B.Matsumoto, ed., Cell Biological Applications of Confocal Microscopy (Academic, 1993), pp. 1–46.
[CrossRef]

S. Inoue, “Foundations of confocal scanned imaging in light microscopy,” in Handbook of Biological Confocal Microscopy, J.Pawley, ed., 3rd ed. (Springer, 2006), pp. 1–19.
[CrossRef]

W. B. Amos, “Achromatic scanning system,” U.S. patent 4,997,242 (5 March 1991).

Photon, Inc., “BeamScan Profiler,” (2009).

J. M. Girkin, “Laser sources for nonlinear microscopy,” in Handbook of Biomedical Nonlinear Optical Microscopy, B.R.Masters and P.T. C.So, eds. (Oxford U. Press, 2008), p. 860.

W. Denk, J. P. Strickler, and W. W. Webb, “Two-photon laser microscopy,” U.S. patent 5,034,613 (23 July 1991).

H. Gross, F. Blechinger, and B. Achtner, Handbook of Optical Systems: Survey of Optical Instruments (Wiley-VCH, 2008), Vol. 4.

G. F. Marshall, Handbook of Optical and Laser Scanning, 2nd ed. (CRC Press, 2004).
[CrossRef]

L. Beiser, Unified Optical Scanning Technology (IEEE, 2003).
[CrossRef]

H.M.Muncheryan, ed., Laser and Optoelectronic Engineering (Hemisphere, 1991).

J. Rietdorf and E. H. K. Stelzer, “Special optical elements,” in Handbook of Biological Confocal Microscopy, J.B.Pawley, ed., 3rd ed. (Springer, 2006), pp. 43–58.
[CrossRef]

E. H. K. Stelzer, “The intermediate optical system of laser-scanning confocal microscopes,” in Handbook of Biological Confocal Microscopy, J.B.Pawley, ed., 3rd ed. (Springer, 2006), pp. 207–220.
[CrossRef]

G. Cox, Optical Imaging Techniques in Cell Biology (CRC, 2007), p. 268.

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Figures (15)

Fig. 1
Fig. 1

Schematic diagram of three simulated and measured scan engine designs: (a) single-mirror engine (RSE), (b) spherical reflector engine (SSE), and (c) parabolic reflector engine (PSE). (d) Describes the scan pattern points analyzed. The investigation points for the PSE and SSE are surrounded by the red and blue rectangles, respectively.

Fig. 2
Fig. 2

Experimental setup of the PSE, which includes four reflective surfaces, a flat mirror, two off-axis parabolic reflectors, and a second flat mirror.

Fig. 3
Fig. 3

Experimentally measured and calculated beam size radii along the propagation axis.

Fig. 4
Fig. 4

(a) Beam propagation after the PSE and SSE relays compared to theoretical beam propagation size. Line 1 is the theoretical beam radius, lines 2 and 3 are the two beam radii after the PSE, and lines 4 and 5 are the two beam radii for the elliptical spot after the SSE. (b) Line 6 is the experimental results for the PSE (line 2) shifted left by the optical path length in the PSE.

Fig. 5
Fig. 5

Radius and divergence of the beam exiting the PSE as a function of the distance between the two parabolic reflectors, M2 and M3. The x axis zero represents the position of the first scan mirror M1 and is 2 m distant from the laser in all cases. The positions of all other mirrors are indicated by marks M2–M4 on the graphs. The output beam with the smallest divergence is achieved for 208.2 mm separation between parabolic reflectors.

Fig. 6
Fig. 6

Measured axial focal shift dependence on parabola separation in the PSE.

Fig. 7
Fig. 7

Experimental cross-sectional images of the laser propagated through the PSE at fifteen scan field points. The spots correspond to the red-labeled region of Fig. 1d. The images in rows represent beam spots deflected by the first scan mirror, and the images in columns are for five tilt angles of the second scan mirror. Scan angle was restricted by the clear aperture of M2.

Fig. 8
Fig. 8

Experimental cross-sectional images of the laser beam at 20 scan field points after the SSE. The spots correspond to the blue-labeled region of Fig. 1d. The images in rows represent beam spots deflected by the first scan mirror, and the images in columns are for five tilt angles of the second scan mirror.

Fig. 9
Fig. 9

(a) Experimental and (b) OSLO-calculated beam spot radii along the major and minor axes, R1 and R2, respectively, over the PSE scan field.

Fig. 10
Fig. 10

(a) Experimental and (b) OSLO-calculated beam spot radii along the major and minor axes, R1 and R2, respectively, over the SSE scan field.

Fig. 11
Fig. 11

Elliptical spot radii R1 and R2 along two main axes at the intermediate image plane. (a) RS: the radii increase evenly in both directions from the central spot. (b) PSE: spot radii for 15 positions showing similar performance to the RS engine. (c) SSE: the beam spot radii for 25 positions showing the effects of spherical aberration.

Fig. 12
Fig. 12

RSE: scan field spot profiles at the intermediate image plane. The spots correspond to the blue-labeled region of Fig. 1d.

Fig. 13
Fig. 13

PSE: spot profiles at chosen positions [the red-labeled region of Fig. 1d] at the intermediate image plane. The images in rows represent beam spots deflected by the first scan mirror, and the images in columns are for five tilt angles of the second scan mirror. Scan angle was restricted by the clear aperture of M2.

Fig. 14
Fig. 14

Beam profile across the chosen scan field at 25 positions. The images in rows represent beam spots deflected by the first scan mirror, and the images in columns are for five tilt angles of the second scan mirror. Spots are more elliptical in the central column, which represents zero deflection of the first scan mirror. Beam circularity for other positions is due to astigmatism created by the two tilted spherical reflector surfaces.

Fig. 15
Fig. 15

Comparison of the field curvature created by the RSE and PSE. (a), (b) Field curvatures in RSE and PSE, respectively, and (c) difference in field curvature between the RSE and PSE.

Tables (3)

Tables Icon

Table 1 Single-Mirror Scan Engine Spot Size Ratio over One Quarter of the Scan Area a

Tables Icon

Table 2 Spot Size Ratio in Parabolic and Spherical Scan Engines a

Tables Icon

Table 3 Relative Field Curvatures for Reference and Parabolic Scan Engines

Equations (1)

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w ( z ) = w 0 [ 1 + ( z z R ) 2 ] 1 / 2 ,

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