CHARACTERIZATION OF FISH SCALE USING 3D PROFILOMETRY 2 4 6 8 1 mm 1 2 3 4 5 6 7 8 mm Prepared by Andrea Novitsky 6 Morgan, Ste156, Irvine CA 92618 P: 949.461.9292 F: 949.461.9232 nanovea.com Today's standard for tomorrow's materials. 214 NANOVEA
INTRODUCTION: The morphology, patterns, and other features of a fish scale are studied using the Nanovea 3D Non-Contact Profilometer. The delicate nature of this biological sample along with its very small and high angled grooves also highlights the importance of the profilometer s non-contact technique. The grooves on the scale are called circuli, and can be studied to estimate the age of the fish, and even distinguish periods of different rates of growth, similar to the rings of a tree. This is very important information for the management of wild fish populations in order to prevent overfishing. IMPORTANCE OF 3D NON CONTACT PROFILOMETER FOR BIOLOGICAL STUDIES Unlike other techniques such as touch probes or interferometry, the 3D Non-Contact Profilometer, using axial chromatism, can measure nearly any surface, sample sizes can vary widely due to open staging and there is no sample preparation needed. Nano through macro range is obtained during surface profile measurement with zero influence from sample reflectivity or absorption, has advanced ability to measure high surface angles and there is no software manipulation of results. Easily measure any material: transparent, opaque, specular, diffusive, polished, rough etc. The technique of the Non Contact Profilometer provides an ideal, broad and user friendly capability to maximize surface studies; along with the benefits of combined 2D & 3D capability. MEASUREMENT OBJECTIVE In this application the Nanovea ST4 Profilometer is used to scan the entire surface of the scale, along with a smaller higher resolution scan in the center of the scale The outer and inner side surface roughness of the scale was also measured for comparison. 2
RESULTS: 3D & 2D Surface Characterization of Outer Scale The 3D View and False Color View of the outer scale show a complex structure similar to a finger print or the rings of a tree. This provides users a straightforward tool to directly observe the surface characterization of the scale from different angles. Various other measurements of the outer scale are shown along with the comparison of the outer and inner side of the scale. 2 4 6 8 1 mm 1 False Color 3 3D View 3 2 25 25 3 2 2 4 15 15 5 1 6 1 5 7 5 8 mm NM..5 1. 1.5 2. 2.5 3. 3.5 4. 4.5 5. 5.5 6. 6.5 7. 7.5 8. mm 1 2 3 4 5 6 7 8 9 1 mm C B A Distances Unit A B C HDist mm 2.156 4.49 1.387 3
Volume 8. Number of islands 75 Threshold 8. Parameters Stat. Value Unit Area Mean.1736 mm² Perimeter Mean 365.3 Orientation Mean 97.43 Volume Mean 2847 ³ Max height Mean 26.6 Step Height 4 L1R1 L2R2 L3R3 L4R4 L5R5 L6R6 2-2 -4..2.4.6.8 1. 1.2 1.4 1.6 1.8 2. mm Parameters Unit Step 1 Step 2 Step 3 Step 4 Step 5 Step 6 Maximum height 57.91 63.46 59.96 57.24 57.5 52.12 4
RESULTS: Surface Roughness Comparison Outer Scale..5 1. 1.5 2. mm..2.4.6.8 1. 1.2 1.4 1.6 1.8 2. mm 12 11 1 9 8 7 6 5 4 3 2 1 Inner Scale 11 1 9 8 7 6 5 4 3 2 1 5
CONCLUSION: In this application, we have shown how the Nanovea 3D Non Contact Profilometer can characterize a fish scale in a variety of ways. The outer and inner surfaces of the scale can be easily distinguished by surface roughness alone, with roughness values of 15.92μm and 1.56μm respectively. Additionally, precise and accurate information can be learned about a fish scale by analyzing the grooves, or circuli, on the outer surface of the scale. The distance of bands of circuli from the center focus was measured, and the height of the circuli was also found to be approximately 58μm high on average. The data shown here represents only a portion of the calculations available in the analysis software. Learn more about the Nanovea Profilometer or Lab Services 6
MEASUREMENT PRINCIPLE: The Chromatic Confocal technique uses a white light source, where light passes through an objective lens with a high degree of chromatic aberration. The refractive index of the objective lens will vary in relation to the wavelength of the light. In effect, each separate wavelength of the incident white light will re-focus at a different distance from the lens (different height). When the measured sample is within the range of possible heights, a single monochromatic point will be focalized to form the image. Due to the confocal configuration of the system, only the focused wavelength will pass through the spatial filter with high efficiency, thus causing all other wavelengths to be out of focus. The spectral analysis is done using a diffraction grating. This technique deviates each wavelength at a different position, intercepting a line of CCD, which in turn indicates the position of the maximum intensity and allows direct correspondence to the Z height position. Unlike the errors caused by probe contact or the manipulative Interferometry technique, Chromatic Confocal technology measures height directly from the detection of the wavelength that hits the surface of the sample in focus. It is a direct measurement with no mathematical software manipulation. This provides unmatched accuracy on the surface measured because a data point is either measured accurately without software interpretation or not at all. The software completes the unmeasured point but the user is fully aware of it and can have confidence that there are no hidden artifacts created by software guessing. Nanovea optical pens have zero influence from sample reflectivity or absorption. Variations require no sample preparation and have advanced ability to measure high surface angles. Capable of large Z measurement ranges. Measure any material: transparent or opaque, specular or diffusive, polished or rough. Measurement includes: Profile Dimension, Roughness Finish Texture, Shape Form Topography, Flatness Warpage Planarity, Volume Area, Step-Height Depth Thickness and many others. A.1
1 z(x, y) dxdy A A 1 A A z2 (x, y)dxdy 1 Sq 3 [1 A A z3 (x, y)dxdy] 1 Sq 4 [1 A A z4 (x, y)dxdy] A.2