Abstract

We present a new imaging method to record multicolor digital holograms from objects emitting fluorescent light. The fluorescent light specific to the emission wavelength of various fluorescent dyes after excitation of three dimensional (3D) objects is recorded on a digital monochrome camera after reflection from a diffractive optical element (DOE). For each wavelength of fluorescent emission, the camera sequentially records three holograms reflected from the DOE, each with a different phase factor of the DOE’s function. The three holograms are superposed in a computer to create a complex valued Fresnel hologram of each fluorescent emission. The holograms for each fluorescent color are further combined in a computer to produce a multicolored fluorescence hologram and 3D color image.

© 2007 Optical Society of America

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References

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  1. J. Garcia-Sucerquia, W. Xu, S. K. Jericho, P. Klages, M. H. Jericho, and H. J. Kreuzer, "Digital in-line holographic microscopy," Appl. Opt. 45, 836-850 (2006).
    [CrossRef] [PubMed]
  2. B. W. Schilling, T.-C. Poon, G. Indebetouw, B. Storrie, K. Shinoda, Y. Suzuki, and M. H. Wu "Three-dimensional holographic fluorescence microscopy," Opt. Lett. 22,1506-1508 (1997).
    [CrossRef]
  3. G. Indebetouw and W. W. Zhong, "Scanning holographic microscopy of three-dimensional fluorescent specimens," J. Opt. Soc. of Am. A 23, 1699-1707 (2006).
    [CrossRef]
  4. J. Rosen, G. Indebetouw, and G. Brooker, "Homodyne scanning holography," Opt. Express 14, 4280-4285 (2006).
    [CrossRef]
  5. J. Rosen and G. Brooker "Digital spatially incoherent Fresnel holography," To be published in Opt. Lett.
  6. I. Yamaguchi and T. Zhang, "Phase-shifting digital holography," Opt. Lett. 22,1268-1269 (1997).
    [CrossRef] [PubMed]
  7. G. Indebetouw and P. Klysubun, "Spatiotemporal digital microholography," J. Opt. Soc. of Am. A 18, 319-325 (2001).
    [CrossRef]
  8. J. Goodman, Introduction to Fourier Optics, 2nd ed. (McGraw-Hill, New York, 1996) 63-95.

2006

2001

G. Indebetouw and P. Klysubun, "Spatiotemporal digital microholography," J. Opt. Soc. of Am. A 18, 319-325 (2001).
[CrossRef]

1997

Brooker, G.

J. Rosen, G. Indebetouw, and G. Brooker, "Homodyne scanning holography," Opt. Express 14, 4280-4285 (2006).
[CrossRef]

J. Rosen and G. Brooker "Digital spatially incoherent Fresnel holography," To be published in Opt. Lett.

Garcia-Sucerquia, J.

Indebetouw, G.

J. Rosen, G. Indebetouw, and G. Brooker, "Homodyne scanning holography," Opt. Express 14, 4280-4285 (2006).
[CrossRef]

G. Indebetouw and W. W. Zhong, "Scanning holographic microscopy of three-dimensional fluorescent specimens," J. Opt. Soc. of Am. A 23, 1699-1707 (2006).
[CrossRef]

G. Indebetouw and P. Klysubun, "Spatiotemporal digital microholography," J. Opt. Soc. of Am. A 18, 319-325 (2001).
[CrossRef]

B. W. Schilling, T.-C. Poon, G. Indebetouw, B. Storrie, K. Shinoda, Y. Suzuki, and M. H. Wu "Three-dimensional holographic fluorescence microscopy," Opt. Lett. 22,1506-1508 (1997).
[CrossRef]

Jericho, M. H.

Jericho, S. K.

Klages, P.

Klysubun, P.

G. Indebetouw and P. Klysubun, "Spatiotemporal digital microholography," J. Opt. Soc. of Am. A 18, 319-325 (2001).
[CrossRef]

Kreuzer, H. J.

Poon, T.-C.

Rosen, J.

J. Rosen, G. Indebetouw, and G. Brooker, "Homodyne scanning holography," Opt. Express 14, 4280-4285 (2006).
[CrossRef]

J. Rosen and G. Brooker "Digital spatially incoherent Fresnel holography," To be published in Opt. Lett.

Schilling, B. W.

Shinoda, K.

Storrie, B.

Suzuki, Y.

Wu, M. H.

Xu, W.

Yamaguchi, I.

Zhang, T.

Zhong, W. W.

G. Indebetouw and W. W. Zhong, "Scanning holographic microscopy of three-dimensional fluorescent specimens," J. Opt. Soc. of Am. A 23, 1699-1707 (2006).
[CrossRef]

Appl. Opt.

J. Opt. Soc. of Am. A

G. Indebetouw and W. W. Zhong, "Scanning holographic microscopy of three-dimensional fluorescent specimens," J. Opt. Soc. of Am. A 23, 1699-1707 (2006).
[CrossRef]

G. Indebetouw and P. Klysubun, "Spatiotemporal digital microholography," J. Opt. Soc. of Am. A 18, 319-325 (2001).
[CrossRef]

Opt. Express

Opt. Lett.

Other

J. Goodman, Introduction to Fourier Optics, 2nd ed. (McGraw-Hill, New York, 1996) 63-95.

J. Rosen and G. Brooker "Digital spatially incoherent Fresnel holography," To be published in Opt. Lett.

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Figures (2)

Fig. 1.
Fig. 1.

Schematic of an incoherent digital hologram recorder. SLM: spatial light modulator; CCD: charge-coupled device; L1, L2, L3 are spherical lenses and F1, F2 are chromatic filters.

Fig. 2.
Fig. 2.

(a). Magnitude and (b) phase of the complex Fresnel hologram of the dice. Digital reconstruction of the non-fluorescence hologram: (c) at the face of the red-dots on the die, and (d) at the face of the green dots on the die. (e) Magnitude and (f) phase of the complex Fresnel hologram of the red dots. Digital reconstruction of the red fluorescence hologram: (g) at the face of the red-dots on the die, and (h) at the face of the green dots on the die. (i) Magnitude and (j) phase of the complex Fresnel hologram of the green dots. Digital reconstruction of the green fluorescence hologram: (k) at the face of the red-dots on the die, and (l) at the face of the green dots on the die. Compositions of Figs 2(c), 2(g) and 2(k) and Figs 2(d), 2(h) and 2(l) are depicted in Figs. 2(m) and 2(n), respectively.

Equations (9)

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R x D y D = 1 2 + 1 2 exp [ i π λ a ( x D 2 + y D 2 ) i θ ] ,
d o ( z s ) ( a d 2 ) ( f 3 f 2 ) 2 + M ¯ A z s = ( a d 2 ) ( f 3 f 2 ) 2 + ( f 3 a f 2 f 1 ) 2 z s
= [ ( r 2 f 1 λ N ) 2 z s + r 2 λ N d 2 ] ( f 3 f 2 ) 2 ,
I p x y C 1 + exp { i π λ d o ( z s ) [ ( x M ¯ T x s ) 2 + ( y M ¯ T y s ) 2 ] i θ } 2 ,
H x y = A ( C + g x s y s z s exp { i π λ d o ( z s ) [ ( x M ̅ T x s ) 2 + ( y M ̅ T y s ) 2 ] + i θ } dx s dy s dz s
+ g x s y s z s exp { i π λ d o ( z s ) [ ( x M ̅ T x s ) 2 + ( y M ̅ T y s ) 2 ] i θ } dx s dy s dz s ) .
H F x y = H 1 x y [ exp ( i θ 3 ) exp ( i θ 2 ) ] + H 2 x y [ exp ( i θ 1 ) exp ( i θ 3 ) ]
+ H 3 x y [ exp ( i θ 2 ) exp ( i θ 1 ) ]
= g x s y s z s exp { i π λ d o ( z s ) [ ( x M ¯ T x s ) 2 + ( y M ¯ T x s ) 2 ] } dx s dy s dz s ,

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