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Imaging quality assessment of multi-modal miniature microscope

Junwon Lee, Jeremy D. Rogers, Michael R. Descour, Elizabeth Hsu, Jesse S. Aaron, Konstantin Sokolov, and Rebecca R. Richards-Kortum

Open Access Open Access

Abstract

We are developing a multi-modal miniature microscope (4M device) to image morphology and cytochemistry in vivo and provide better delineation of tumors. The 4M device is designed to be a complete microscope on a chip, including optical, micro-mechanical, and electronic components. It has advantages such as compact size and capability for microscopic-scale imaging. This paper presents an optics-only prototype 4M device, the very first imaging system made of sol-gel material. The micro-optics used in the 4M device has a diameter of 1.3 mm. Metrology of the imaging quality assessment of the prototype device is presented. We describe causes of imaging performance degradation in order to improve the fabrication process. We built a multi-modal imaging test-bed to measure first-order properties and to assess the imaging quality of the 4M device. The 4M prototype has a field of view of 290 µm in diameter, a magnification of -3.9, a working distance of 250 µm and a depth of field of 29.6±6 µm. We report the modulation transfer function (MTF) of the 4M device as a quantitative metric of imaging quality. Based on the MTF data, we calculated a Strehl ratio of 0.59. In order to investigate the cause of imaging quality degradation, the surface characterization of lenses in 4M devices is measured and reported. We also imaged both polystyrene microspheres similar in size to epithelial cell nuclei and cervical cancer cells. Imaging results indicate that the 4M prototype can resolve cellular detail necessary for detection of precancer.

©2003 Optical Society of America

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Figures (10)
Fig. 1.
Fig. 1. Conceptual geometry of 4M device. Every component is mounted on one silicon substrate [a.k.a., the Micro Optical Table, MOT] including an imaging sensor and an illumination system. The substrate dimensions are 13mm (L)×10 mm (W). Blank lines are geometrical rays traced non-sequentially in the 4M device.

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Fig. 2.
Fig. 2. Lens Design of 4M Device. 4M devices consist of one objective lens from Edmund industrial optics and three hybrid lenses that are fabricated by photo-lithography. The object is assumed to be in water.

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Fig. 3.
Fig. 3. Magnified view of prototype 4M-device. (a) Front view of 4M device. Part (b) shows a side view of a complete, “optics-only” 4M device. An objective lens is embedded in the MOT.

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Fig. 4.
Fig. 4. Schematic diagram of multi-mode imaging test-bed: (a) Top view of entire system. It shows two fibers for two different illumination modes: Fiber A for trans-illumination mode and Fiber B for epi-illumination mode. (b) Side view of region inside the blue box in (a). It shows trans-illumination mode.

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Fig. 5.
Fig. 5. Actual multi-modal imaging test-bed in figure (a). The 3-axis actuators shown within the red oval are used to control the object, the 4M device and the trans-illumination system. The 4M device is located inside the oval. Figure (b) provides the magnified view of the 4M device being used with an imaging chamber used to hold cells. An object, usually cells in liquid media, is contained in sample holder underneath 4M device.

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Fig. 6.
Fig. 6. MTF curve for the 4M device reported in this paper (Green Line). The black dots are the measured CTF data and the red line is the interpolated CTF. The blue line above other lines is the diffraction limited MTF of a 4M device with NA=0.15.

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Fig. 7.
Fig. 7. Zernike coefficients of objective lens and 3 hybrid lenses: (a) objective lens, (b) hybrid lens 1, (c) hybrid lens 2, and (d) hybrid lens 3. Red triangles are correct Zernike coefficients and black rectangles are reconstructed Zernike coefficients from measured surface profiles.

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Fig. 8.
Fig. 8. Contour plots of objective lens and 3 hybrid lenses: (a) objective lens, (b) hybrid lens 1, (c) hybrid lens 2, and (d) hybrid lens 3. It shows irregularity of contours and non-rotationally symmetric shape which are main causes of imaging performance degradation. The FOV is 224 µm×295 µm in plot (a) and is 456 µm×600 µm in plot (b), (c) and (d).

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Fig. 9.
Fig. 9. Images of polystyrene beads in trans-illumination mode in figure (a) and (b). As expected, the negative contrasts (dark ring patterns) are visible around polystyrene beads in all parts of the Figure. Parts (c) and (d) show that two polystyrene beads can adhere to each other and still be clearly resolvable. Parts (b) and (d) are the magnified sections indicated by red boxes in Parts (a) and (c), respectively.

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Fig. 10.
Fig. 10. Images of cervical-cancer cells (SiHa) in trans-illumination mode. The cell membrane is shown due to small change of refractive index between cells and media (a). Figure (b) shows the magnified view of region inside red box in figure (a). Within the cell images in figure (b), the unstained nuclei are visible.

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Tables (6)

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Table 1. First order design of 4M devices

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Table 2. Lens prescription data of 4M devices

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Table 3. RMS wavefront error in waves of three hybrid lenses used in 4M device

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Table 4. First-order properties of 4M device.

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Table 5. Surface roughness of objective lens and hybrid lenses. Surface roughness is defined as RMS error between measured surface data and designed surface data. The unit is nano-meter. FOV 1 has 240 µm×180 µm of FOV and FOV 2 has 610 µm×460 µm of FOV.

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Table 6. Radii of curvature of objective lens and hybrid lenses in mm.

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Equations (6)

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

s ( ξ o ) = 4 π ∑ n = 0 ∞ ( − 1 ) n ( 2 n + 1 ) m [ ( 2 n + 1 ) ξ o ] ,
m ( ξ o ) = π 4 ∑ n = 0 ∞ B 2 n + 1 s [ ( 2 n + 1 ) ξ o ] ( 2 n + 1 ) .
m ( ξ o ) = π 4 ∑ n = 0 N B 2 n + 1 s [ ( 2 n + 1 ) ξ o ] ( 2 n + 1 ) for ξ c ( 2 N + 3 ) < v ≤ ξ c ( 2 N + 1 ) .
Contrast = I max − I min I max + I min .
S ( x , y ) = ∑ i = 1 i max c i Z i ( x , y ) ,
r = ρ 2 2 ( 2 c 3 − 6 c 8 + 12 c 15 … ) ,
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