Abstract

We have developed a high-speed confocal laser microscope. A microlens-array disk set in front of a pinhole-array disk improved optical efficiency more than ten times compared with that of conventional Nipkow confocal microscopy. This new microscope achieves a high-speed measurement of 1 frame/ms. We expect that it will be used for measuring biological and industrial active samples.

© 2002 Optical Society of America

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References

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  1. R. C. Mellors, R. Silver, “A microfluorometric scanner for the differential detection of cells,” Science 114, 356–361 (1951).
    [CrossRef] [PubMed]
  2. T. Tanaami, Y. Sugiyama, K. Mikuriya, “High-speed confocal laser microscopy,” Yokogawa Tech. Rep. 9, English ed. (Yokogawa Electric Corporation, Tokyo, Japan, 1994), pp. 7–10.
  3. A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, I. Umeda, “High-speed confocal fluorescent microscopy using a Nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimages 4(2), 57–62 (1996).
  4. C. Genka, H. Ishida, K. Ichimori, Y. Hirota, T. Tanaami, H. Nakazawa, “Visualization of biphasic Ca2+ diffusion from cytesol to nucleus in contracting adult rat cardiac myocytes with an ultra-fast confocal imaging system,” Cell Calcium 25, 199–208 (1999).
    [CrossRef] [PubMed]

1999 (1)

C. Genka, H. Ishida, K. Ichimori, Y. Hirota, T. Tanaami, H. Nakazawa, “Visualization of biphasic Ca2+ diffusion from cytesol to nucleus in contracting adult rat cardiac myocytes with an ultra-fast confocal imaging system,” Cell Calcium 25, 199–208 (1999).
[CrossRef] [PubMed]

1996 (1)

A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, I. Umeda, “High-speed confocal fluorescent microscopy using a Nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimages 4(2), 57–62 (1996).

1951 (1)

R. C. Mellors, R. Silver, “A microfluorometric scanner for the differential detection of cells,” Science 114, 356–361 (1951).
[CrossRef] [PubMed]

Abe, M.

A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, I. Umeda, “High-speed confocal fluorescent microscopy using a Nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimages 4(2), 57–62 (1996).

Genka, C.

C. Genka, H. Ishida, K. Ichimori, Y. Hirota, T. Tanaami, H. Nakazawa, “Visualization of biphasic Ca2+ diffusion from cytesol to nucleus in contracting adult rat cardiac myocytes with an ultra-fast confocal imaging system,” Cell Calcium 25, 199–208 (1999).
[CrossRef] [PubMed]

Hirota, Y.

C. Genka, H. Ishida, K. Ichimori, Y. Hirota, T. Tanaami, H. Nakazawa, “Visualization of biphasic Ca2+ diffusion from cytesol to nucleus in contracting adult rat cardiac myocytes with an ultra-fast confocal imaging system,” Cell Calcium 25, 199–208 (1999).
[CrossRef] [PubMed]

Ichihara, A.

A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, I. Umeda, “High-speed confocal fluorescent microscopy using a Nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimages 4(2), 57–62 (1996).

Ichimori, K.

C. Genka, H. Ishida, K. Ichimori, Y. Hirota, T. Tanaami, H. Nakazawa, “Visualization of biphasic Ca2+ diffusion from cytesol to nucleus in contracting adult rat cardiac myocytes with an ultra-fast confocal imaging system,” Cell Calcium 25, 199–208 (1999).
[CrossRef] [PubMed]

Ishida, H.

C. Genka, H. Ishida, K. Ichimori, Y. Hirota, T. Tanaami, H. Nakazawa, “Visualization of biphasic Ca2+ diffusion from cytesol to nucleus in contracting adult rat cardiac myocytes with an ultra-fast confocal imaging system,” Cell Calcium 25, 199–208 (1999).
[CrossRef] [PubMed]

Isozaki, K.

A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, I. Umeda, “High-speed confocal fluorescent microscopy using a Nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimages 4(2), 57–62 (1996).

Kosugi, Y.

A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, I. Umeda, “High-speed confocal fluorescent microscopy using a Nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimages 4(2), 57–62 (1996).

Mellors, R. C.

R. C. Mellors, R. Silver, “A microfluorometric scanner for the differential detection of cells,” Science 114, 356–361 (1951).
[CrossRef] [PubMed]

Mikuriya, K.

A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, I. Umeda, “High-speed confocal fluorescent microscopy using a Nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimages 4(2), 57–62 (1996).

T. Tanaami, Y. Sugiyama, K. Mikuriya, “High-speed confocal laser microscopy,” Yokogawa Tech. Rep. 9, English ed. (Yokogawa Electric Corporation, Tokyo, Japan, 1994), pp. 7–10.

Nakazawa, H.

C. Genka, H. Ishida, K. Ichimori, Y. Hirota, T. Tanaami, H. Nakazawa, “Visualization of biphasic Ca2+ diffusion from cytesol to nucleus in contracting adult rat cardiac myocytes with an ultra-fast confocal imaging system,” Cell Calcium 25, 199–208 (1999).
[CrossRef] [PubMed]

Silver, R.

R. C. Mellors, R. Silver, “A microfluorometric scanner for the differential detection of cells,” Science 114, 356–361 (1951).
[CrossRef] [PubMed]

Sugiyama, Y.

A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, I. Umeda, “High-speed confocal fluorescent microscopy using a Nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimages 4(2), 57–62 (1996).

T. Tanaami, Y. Sugiyama, K. Mikuriya, “High-speed confocal laser microscopy,” Yokogawa Tech. Rep. 9, English ed. (Yokogawa Electric Corporation, Tokyo, Japan, 1994), pp. 7–10.

Tanaami, T.

C. Genka, H. Ishida, K. Ichimori, Y. Hirota, T. Tanaami, H. Nakazawa, “Visualization of biphasic Ca2+ diffusion from cytesol to nucleus in contracting adult rat cardiac myocytes with an ultra-fast confocal imaging system,” Cell Calcium 25, 199–208 (1999).
[CrossRef] [PubMed]

A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, I. Umeda, “High-speed confocal fluorescent microscopy using a Nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimages 4(2), 57–62 (1996).

T. Tanaami, Y. Sugiyama, K. Mikuriya, “High-speed confocal laser microscopy,” Yokogawa Tech. Rep. 9, English ed. (Yokogawa Electric Corporation, Tokyo, Japan, 1994), pp. 7–10.

Umeda, I.

A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, I. Umeda, “High-speed confocal fluorescent microscopy using a Nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimages 4(2), 57–62 (1996).

Bioimages (1)

A. Ichihara, T. Tanaami, K. Isozaki, Y. Sugiyama, Y. Kosugi, K. Mikuriya, M. Abe, I. Umeda, “High-speed confocal fluorescent microscopy using a Nipkow scanner with microlenses for 3-d imaging of single fluorescent molecule in real time,” Bioimages 4(2), 57–62 (1996).

Cell Calcium (1)

C. Genka, H. Ishida, K. Ichimori, Y. Hirota, T. Tanaami, H. Nakazawa, “Visualization of biphasic Ca2+ diffusion from cytesol to nucleus in contracting adult rat cardiac myocytes with an ultra-fast confocal imaging system,” Cell Calcium 25, 199–208 (1999).
[CrossRef] [PubMed]

Science (1)

R. C. Mellors, R. Silver, “A microfluorometric scanner for the differential detection of cells,” Science 114, 356–361 (1951).
[CrossRef] [PubMed]

Other (1)

T. Tanaami, Y. Sugiyama, K. Mikuriya, “High-speed confocal laser microscopy,” Yokogawa Tech. Rep. 9, English ed. (Yokogawa Electric Corporation, Tokyo, Japan, 1994), pp. 7–10.

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Figures (8)

Fig. 1
Fig. 1

Microlens- and Nipkow-disk scanning confocal microscope.

Fig. 2
Fig. 2

Diagrams to explain the arrangement of pinholes.

Fig. 3
Fig. 3

System construction. PC, personal computer.

Fig. 4
Fig. 4

Resolution along the optical axis.

Fig. 5
Fig. 5

Noise estimation of camera.

Fig. 6
Fig. 6

Noise estimation of camera and image intensifier.

Fig. 7
Fig. 7

High-speed confocal image; encoder of linear actuator (pitch, 0.17 mm).

Fig. 8
Fig. 8

Fluorescent images of ventricular cell from adult rat (1 ms/frame).

Tables (2)

Tables Icon

Table 1 Typical Features of Confocal Scanners

Tables Icon

Table 2 Comparison of Pinhole Patterns

Equations (7)

Equations on this page are rendered with MathJax. Learn more.

r=ro+θma/2π,
L=00 rdθ=roθ+maθ22×2π.
θ=2πma-ro+ro2+maLπ1/2.
L=ia,
ri=ro+θima/2π,
θ=2πma-ro+ro2+ima2π1/2.
Nshot=2q Pin B1/2,

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