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An optical coherence microscope for 3-dimensional imaging in developmental biology

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Abstract

An optical coherence microscope (OCM) has been designed and constructed to acquire 3-dimensional images of highly scattering biological tissue. Volume-rendering software is used to enhance 3-D visualization of the data sets. Lateral resolution of the OCM is 5 µm (FWHM), and the depth resolution is 10 µm (FWHM) in tissue. The design trade-offs for a 3-D OCM are discussed, and the fundamental photon noise limitation is measured and compared with theory. A rotating 3-D image of a frog embryo is presented to illustrate the capabilities of the instrument.

©2000 Optical Society of America

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Supplementary Material (2)

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Media 2: MOV (386 KB)     

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Figures (6)

Fig. 1.
Fig. 1. Optical schematic of the optical coherence microscope.
Fig. 2.
Fig. 2. Phase dependence of the output fringe signal.
Fig. 3.
Fig. 3. Maximum powers in the output fringe signal at the first two harmonics of the piezo-driving voltage.
Fig. 4.
Fig. 4. Photodetector noise as a function of reference beam power. A spectrum analyzer was used to measure the photodetector noise (solid circles) at 122.5 KHz with a bandwidth BW=3 KHz. The theoretical curve (solid line) is calculated using this value of the bandwidth in equation (6).
Fig. 5.
Fig. 5. OCM signal-to-noise ratio as a function of reference beam power. The solid line is calculated from equation (8) with no adjustable parameters. The dashed line includes an additional multiplicative constant of 0.40.
Fig. 6.
Fig. 6. First frame of the movie of a frog embryo at stage 41 of development. The movie is provided in two sizes: a 2.46 MB file and also a 385 KB version with slightly lower resolution but faster download time.

Equations (10)

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I out = I 0 cos ( α sin ω t + ϕ )
P 1 = 2 I 0 2 J 1 2 ( α ) sin 2 ϕ
P 2 = 2 I 0 2 J 2 2 ( α ) cos 2 ϕ
Var ( n ) = < n > + < n > 2 N
Var ( n ) = 2 [ < n > 2 + 1 N ( < n > 2 ) 2 ] = < n > + 1 2 < n > 2 N
V rms noise = g [ ( R · NEP ) 2 · BW ] + [ 2 η e 2 BW P ref E ν ] + [ ( η e P ref E ν ) 2 τ coh BW ]
V rms signal = 1.31 η eg P ref P samp E ν
SNR = 1.31 η e P ref P samp E ν [ ( R . NEP ) 2 . BW ] + [ 2 η e 2 BW P ref E ν ] + [ ( η e P ref E ν ) 2 τ coh BW ]
[ Pixel RGB ] = n α n * [ Voxel RGB ] n
α n = 1
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