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Experiments in Fluids
Erschienen in: Experiments in Fluids 1/2004

01.07.2004 | Original

Optically sliced micro-PIV using confocal laser scanning microscopy (CLSM)

verfasst von: Jae Sung Park, Chang Kyoung Choi, Kenneth D. Kihm

Erschienen in: Experiments in Fluids | Ausgabe 1/2004

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Abstract

Optically sliced microscopic-particle image velocimetry (micro-PIV) is developed using confocal laser scanning microscopy (CLSM). The developed PIV system shows a unique optical slicing capability allowing true depth-wise resolved micro-PIV vector field mapping. A comparative study between CLSM micro-PIV and conventional epi-fluorescence micro-PIV is presented. Both techniques have been applied to the creeping Poiseuille flows in two different microtubes of 99-μm (Re=0.00275) and 516-μm ID diameters (Re=0.021), which are respectively imaged by a 40×-0.75NA objective with an estimated 2.8-μm optical slice thickness, and by a 10×-0.30NA objective with a 26.7-μm slicing. Compared to conventional micro-PIV, CLSM micro-PIV consistently shows significantly improved particle image contrasts, definitions, and measured flow vector fields agreeing more accurately with predictions based on the Poiseuille flow fields. The data improvement due to the optical slicing of CLSM micro-PIV is more pronounced with higher magnification imaging with higher NA objectives for a smaller microtube.
Vorheriger Artikel Analysis of the wake–mixing-layer interaction using multiple plane PIV and 3D classical POD
Nächster Artikel Spatial perturbation of a wing-tip vortex using pulsed span-wise jets

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Fußnoten
1
This is also called a “far-field” diffraction condition, which is defined as R>a 2/λ, where R is the smaller of the two distances from the particle to the objective lens and the objective lens to the imaging detector, a is the particle radius, and λ is the wavelength in the medium. For typical conditions for micro-PIV, R~1 mm, a~200 nm, and λ~500 nm, the inequality is well satisfied by a ratio of greater than 12,000.
 
2
Numerical aperture, NA, is defined as \(NA \equiv n_{i} \sin \theta _{{\max }} \), where n i is the refractive index of the immersing medium (air, water, oil, etc.) adjacent to the objective lens, and θ max is the half-angle of the maximum cone of the light apertured by the lens.
 
3
Modified pinhole diameter, PD, is defined as pinhole diameter/magnification, with the pinhole diameter measured in μm.
 
4
Airy unit, \({\text{AU}} \equiv {1.22\lambda _{{{\text{ex}}}} } \mathord{\left/ {\vphantom {{1.22\lambda _{{{\text{ex}}}} } {NA}}} \right. \kern-\nulldelimiterspace} {NA} \), with λ ex being the fluorescent excitation wavelength.
 
5
The mean wavelength is defined as, \(\ifmmode\expandafter\bar\else\expandafter\=\fi{\lambda } = {\sqrt 2 }\frac{{\lambda _{{{\text{ex}}}} \cdot \lambda _{{{\text{em}}}} }}{{{\sqrt {\lambda _{{{\text{ex}}}} ^{2} + \lambda _{{{\text{em}}}} ^{2} } }}} \).
 
6
The reduced apparent depth, h a, is derived as h/n, based on Snell’s law of refraction (Hecht 2002). Strictly speaking, the analysis assumes a planar interface and zero ray-incident angle, thus, for the case of a circular microtube, it is only valid along the centerline (refer to Fig. 9).
 
7
The spatial uncertainty of the imaging planes is estimated as an rms of the one-half of the micro-stage reading resolution, 0.5 μm, and the uncertainty level for identifying the top-end point is estimated to be approximately identical to the image depth-of-field (DOF), 0.92 µm for 40× and 5.72 µm for 10× magnification, for the conventional microscope. Note that the values of DOF for CLSM are 0.63 µm and 4.66 µm for 40× and 10× magnification, respectively.
 
8
The background noise from the off-focus particle images can be reduced to an acceptable level by limiting the PIV measurement depth to a base-cut level where the field-wide-averaged image intensity reaches one-tenth of the maximum in-focus image intensity (Meinhart et al. 2000).
 
9
The actual recorded image of a seed particle on the CCD is a convolution of the geometric particle image, Md p, with the FPS, d s, of the recording optics. Approximating both of the geometric and diffraction-limited images as Gaussian functions, the image diameter, d e, can be expressed as (Born and Wolf 1999) \(d_{{\text{e}}} = {\left[ {M^{2} d_{{\text{p}}} ^{2} + d_{{\text{s}}} ^{2} } \right]}^{{1/2}} \) where d e is the effective particle diameter in the CCD, M is the magnification of the microscope, d p is the particle diameter, and d s is the characteristic diameter of the PSF. For magnifications much larger than unity, the diameter of the diffraction-limited PSF, in the image plane, is given by \(d_{{\text{s}}} = 2.44M\frac{\lambda }{{2NA}} \) where NA is the numerical aperture and λ is the wavelength of light.
 
10
Optical path length (OPL) is defined as the local medium thickness multiplied by the local refractive index.
 
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Metadaten
Titel
Optically sliced micro-PIV using confocal laser scanning microscopy (CLSM)
verfasst von
Jae Sung Park
Chang Kyoung Choi
Kenneth D. Kihm
Publikationsdatum
01.07.2004
Verlag
Springer-Verlag
Erschienen in
Experiments in Fluids / Ausgabe 1/2004
Print ISSN: 0723-4864
Elektronische ISSN: 1432-1114
DOI
https://doi.org/10.1007/s00348-004-0790-6

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