Abstract

The multiple imaging axis microscope (MIAM) is a wide-field optical microscope that observes a sample simultaneously from multiple directions without requiring the sample to be rotated or tilted. The prototype is capable of high-resolution imaging of the interior of a 300µm-diameter sample consisting of fluorescent microbeads suspended in an agarose gel. Compared with a single-axis system, the MIAM can achieve a reduction of the axial point-spread function elongation by a factor of 5.8 and a 3.5-fold improvement in volume resolution by simple linear image combination techniques.

© 2003 Optical Society of America

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    [CrossRef]

1999 (2)

S. Grill and E. H. K. Stelzer, J. Opt. Soc. Am. A 16, 2658 (1999).
[CrossRef]

P. J. Verveer, M. J. Gemkow, and T. M. Jovin, J. Microsc. 193, 50 (1999).
[CrossRef]

1998 (1)

E. H. K. Stelzer, J. Microsc. 189, 15 (1998).
[CrossRef]

1997 (3)

K. Sätzler and R. Elis, Bioimaging 5, 171 (1997).
[CrossRef]

S. Kikuchi, K. Sonobe, S. Mashiko, Y. Hiraoka, and N. Ohyama, Opt. Commun. 138, 21 (1997).
[CrossRef]

K. G. Larkin, M. A. Oldfield, and H. Klemm, Opt. Commun. 139, 99 (1997).
[CrossRef]

1994 (3)

J. Bradl, M. Hausmann, B. Schneider, B. Rinke, and C. Cremer, J. Microsc. 175, 211 (1994).
[CrossRef]

S. Kikuchi, K. Sonobe, L. S. Didharta, and N. Ohyama, Opt. Commun. 107, 432 (1994).
[CrossRef]

E. H. K. Stelzer and S. Lindek, Opt. Commun. 111, 536 (1994).
[CrossRef]

1992 (2)

1990 (1)

W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 4951 (1990).
[CrossRef]

1989 (1)

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef]

Agard, D. A.

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef]

Bradl, J.

J. Bradl, M. Hausmann, B. Schneider, B. Rinke, and C. Cremer, J. Microsc. 175, 211 (1994).
[CrossRef]

Cremer, C.

J. Bradl, M. Hausmann, B. Schneider, B. Rinke, and C. Cremer, J. Microsc. 175, 211 (1994).
[CrossRef]

Denk, W.

W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 4951 (1990).
[CrossRef]

Didharta, L. S.

S. Kikuchi, K. Sonobe, L. S. Didharta, and N. Ohyama, Opt. Commun. 107, 432 (1994).
[CrossRef]

Elis, R.

K. Sätzler and R. Elis, Bioimaging 5, 171 (1997).
[CrossRef]

Gemkow, M. J.

P. J. Verveer, M. J. Gemkow, and T. M. Jovin, J. Microsc. 193, 50 (1999).
[CrossRef]

Gottesfeld Brown, L.

L. Gottesfeld Brown, ACM Comput. Surv. 24, 325 (1992).
[CrossRef]

Grill, S.

Hausmann, M.

J. Bradl, M. Hausmann, B. Schneider, B. Rinke, and C. Cremer, J. Microsc. 175, 211 (1994).
[CrossRef]

Hell, S.

Hiraoka, Y.

S. Kikuchi, K. Sonobe, S. Mashiko, Y. Hiraoka, and N. Ohyama, Opt. Commun. 138, 21 (1997).
[CrossRef]

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef]

Jovin, T. M.

P. J. Verveer, M. J. Gemkow, and T. M. Jovin, J. Microsc. 193, 50 (1999).
[CrossRef]

Kikuchi, S.

S. Kikuchi, K. Sonobe, S. Mashiko, Y. Hiraoka, and N. Ohyama, Opt. Commun. 138, 21 (1997).
[CrossRef]

S. Kikuchi, K. Sonobe, L. S. Didharta, and N. Ohyama, Opt. Commun. 107, 432 (1994).
[CrossRef]

Klemm, H.

K. G. Larkin, M. A. Oldfield, and H. Klemm, Opt. Commun. 139, 99 (1997).
[CrossRef]

Larkin, K. G.

K. G. Larkin, M. A. Oldfield, and H. Klemm, Opt. Commun. 139, 99 (1997).
[CrossRef]

Lindek, S.

E. H. K. Stelzer and S. Lindek, Opt. Commun. 111, 536 (1994).
[CrossRef]

Mashiko, S.

S. Kikuchi, K. Sonobe, S. Mashiko, Y. Hiraoka, and N. Ohyama, Opt. Commun. 138, 21 (1997).
[CrossRef]

Ohyama, N.

S. Kikuchi, K. Sonobe, S. Mashiko, Y. Hiraoka, and N. Ohyama, Opt. Commun. 138, 21 (1997).
[CrossRef]

S. Kikuchi, K. Sonobe, L. S. Didharta, and N. Ohyama, Opt. Commun. 107, 432 (1994).
[CrossRef]

Oldfield, M. A.

K. G. Larkin, M. A. Oldfield, and H. Klemm, Opt. Commun. 139, 99 (1997).
[CrossRef]

Rinke, B.

J. Bradl, M. Hausmann, B. Schneider, B. Rinke, and C. Cremer, J. Microsc. 175, 211 (1994).
[CrossRef]

Sätzler, K.

K. Sätzler and R. Elis, Bioimaging 5, 171 (1997).
[CrossRef]

Schneider, B.

J. Bradl, M. Hausmann, B. Schneider, B. Rinke, and C. Cremer, J. Microsc. 175, 211 (1994).
[CrossRef]

Sedat, J. W.

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef]

Shaw, P. J.

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef]

Sonobe, K.

S. Kikuchi, K. Sonobe, S. Mashiko, Y. Hiraoka, and N. Ohyama, Opt. Commun. 138, 21 (1997).
[CrossRef]

S. Kikuchi, K. Sonobe, L. S. Didharta, and N. Ohyama, Opt. Commun. 107, 432 (1994).
[CrossRef]

Stelzer, E. H. K.

Strickler, J. H.

W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 4951 (1990).
[CrossRef]

Verveer, P. J.

P. J. Verveer, M. J. Gemkow, and T. M. Jovin, J. Microsc. 193, 50 (1999).
[CrossRef]

Webb, W. W.

W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 4951 (1990).
[CrossRef]

ACM Comput. Surv. (1)

L. Gottesfeld Brown, ACM Comput. Surv. 24, 325 (1992).
[CrossRef]

Bioimaging (1)

K. Sätzler and R. Elis, Bioimaging 5, 171 (1997).
[CrossRef]

Biophys. J. (1)

P. J. Shaw, D. A. Agard, Y. Hiraoka, and J. W. Sedat, Biophys. J. 55, 101 (1989).
[CrossRef]

J. Microsc. (3)

P. J. Verveer, M. J. Gemkow, and T. M. Jovin, J. Microsc. 193, 50 (1999).
[CrossRef]

J. Bradl, M. Hausmann, B. Schneider, B. Rinke, and C. Cremer, J. Microsc. 175, 211 (1994).
[CrossRef]

E. H. K. Stelzer, J. Microsc. 189, 15 (1998).
[CrossRef]

J. Opt. Soc. Am. A (2)

Opt. Commun. (4)

E. H. K. Stelzer and S. Lindek, Opt. Commun. 111, 536 (1994).
[CrossRef]

S. Kikuchi, K. Sonobe, L. S. Didharta, and N. Ohyama, Opt. Commun. 107, 432 (1994).
[CrossRef]

S. Kikuchi, K. Sonobe, S. Mashiko, Y. Hiraoka, and N. Ohyama, Opt. Commun. 138, 21 (1997).
[CrossRef]

K. G. Larkin, M. A. Oldfield, and H. Klemm, Opt. Commun. 139, 99 (1997).
[CrossRef]

Science (1)

W. Denk, J. H. Strickler, and W. W. Webb, Science 248, 4951 (1990).
[CrossRef]

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Figures (3)

Fig. 1
Fig. 1

(a) Objective lenses OL1OL4 in the tetrahedral geometry, with corresponding focal planes FP1FP4 intersecting in the sample region. Components of arm 1 are emphasized. (b) Simplified schematic of a single MIAM arm. D, dichroic beam splitter; EF, emission filter; TL, tube lens; CCD, camera; C, collection lens; IF, illumination fiber.

Fig. 2
Fig. 2

Maximum-value projections, 200-nm fluorescent beads. (a) Large field-of-view image, CCD 1. (b), (c), (d), and (e) Cropped images of CCDs 1, 2, 3, and 4, respectively. Fusions of data from (b)–(e) by (f) arithmetic average and (g) weighted average are also shown.

Fig. 3
Fig. 3

Line scans through 3D stacks of a 200-nm bead. Dashed curves CCD 1. Solid curves, weighted-average fusion image. Inset, projections (perpendicular to optical axis of CCD 1) from CCD 1 and the fusion data set; arrows indicate line scan directions.

Tables (1)

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Table 1 Resolution and Gaina

Equations (2)

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If,a=1NjNIj;
If,w=FT-1jNwjI˜j, wj=I˜j/jNI˜k, I˜j=FTIj.

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