Name: a millimeter scale flexural testing system for measuring the mechanical properties of marine sponge spicules
Uploaded: 2017-10-11T15:00:00
Description: Watch this Scientific Journal Video about A Millimeter Scale Flexural Testing System for Measuring the Mechanical Properties of Marine Sponge Spicules at JoVE.com

यह काम राष्ट्रीय विज्ञान फाउंडेशन द्वारा समर्थित किया गया था [सामग्री और संरचनाओं कार्यक्रम के यांत्रिकी, अनुदान संख्या १५६२६५६]; और अमेरिकी सोसायटी ऑफ मैकेनिकल इंजीनियर्स [Haythornthwaite यंग जांचकर्ता पुरस्कार] ।

1. विधानसभा और संरेखण <ol> <li> एक ब्रैकट चुनें जिसका कड़ा इरादा प्रयोग के लिए उपयुक्त है । एल. पी. #4 का उपयोग कर ब्रैकट को अनुलग्न करें-४० सॉकेट सिर कैप शिकंजा (SHCSs) (see < सुदृढ वर्ग = "xfig" > चित्रा ३ अ ). एल. प?...

<ol>
	<li>Wegst, U. G., Bai, H., Saiz, E., Tomsia, A. P., Ritchie, R. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bioinspired+structural+materials.">Bioinspired structural materials.</a> Nat. Mater. 14 (1), 23-36 (2015).</li><li>Meyers, M. A., McKittrick, J., Chen, P. Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structural+biological+materials:+critical+mechanics-materials+connections.">Structural biological materials: critical mechanics-materials connections.</a> Science. 339 (6121), 773-779 (2013).</li><li>Bodde, S. G., Meyers, M. A., McKittrick, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Correlation+of+the+mechanical+and+structural+properties+of+cortical+rachis+keratin+of+rectrices+of+the+Toco+Toucan+(Ramphastos+toco).">Correlation of the mechanical and structural properties of cortical rachis keratin of rectrices of the Toco Toucan (Ramphastos toco).</a> J. Mech. Behav. Biomed. Mater. 4 (5), 723-732 (2011).</li><li>Gibson, L. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+hierarchical+structure+and+mechanics+of+plant+materials.">The hierarchical structure and mechanics of plant materials.</a> J. R. Soc. Interface. , (2012).</li><li>Monn, M. A., Kesari, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+structure-property+connection+in+the+skeletal+elements+of+the+marine+sponge+Tethya+aurantia+that+guards+against+buckling+instability.">A new structure-property connection in the skeletal elements of the marine sponge Tethya aurantia that guards against buckling instability.</a> Sci. Rep. 7, (2017).</li><li>Monn, M. A., Weaver, J. C., Zhang, T., Aizenberg, J., Kesari, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+functional+insights+into+the+internal+architecture+of+the+laminated+anchor+spicules+of+Euplectella+aspergillum.">New functional insights into the internal architecture of the laminated anchor spicules of Euplectella aspergillum.</a> Proc. Natl. Acad. Sci. U.S.A. 112 (16), 4976-4981 (2015).</li><li>Monn, M. 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Mech. 43 (3), 248-255 (2003).</li><li>Gudlavalleti, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Mechanical testing of solid materials at the micro-scale. , (2002).</li><li>Tohmyoh, H., Ishihara, M., Akanda, M. S., Yamaki, S., Watanabe, T., Iwabuchi, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Accurate+determination+of+the+structural+elasticity+of+human+hair+by+a+small-scale+bending+test.">Accurate determination of the structural elasticity of human hair by a small-scale bending test.</a> J. Biomech. 44 (16), 2833-2837 (2011).</li><li>Waters, J. 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N., Mazzolai, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Structure+and+mechanical+properties+of+Octopus+vulgaris+suckers.">Structure and mechanical properties of Octopus vulgaris suckers.</a> J. R. Soc. Interface. 11 (91), (2014).</li><li>Ehrlich, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Nanostructural+organization+of+naturally+occurring+composites:+Part+I.+Silica-Collagen-based+biocomposites.">Nanostructural organization of naturally occurring composites: Part I. Silica-Collagen-based biocomposites.</a> J. Nanomater. 53, (2008).</li><li>Ehrlich, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mineralization+of+the+meter-long+biosilica+structures+of+glass+sponges+is+templated+on+hydroxylated+collagen.">Mineralization of the meter-long biosilica structures of glass sponges is templated on hydroxylated collagen.</a> Nat. Chem. 2, 1084-1088 (2010).</li><li>Ehrlich, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=First+evidence+of+the+presence+of+chitin+in+skeletons+of+marine+sponges.+Part+II.+Glass+sponges+(Hexactinellida:+Porifera).">First evidence of the presence of chitin in skeletons of marine sponges. Part II. Glass sponges (Hexactinellida: Porifera).</a> J. Exp. 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Mater. 4 (10), 1608-1613 (2016).</li><li>Ehrlich, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Calcite+reinforced+silica-silica+joints+in+the+biocomposite+skeleton+of+deep-sea+glass+sponges.">Calcite reinforced silica-silica joints in the biocomposite skeleton of deep-sea glass sponges.</a> Adv. Funct. Mater. 21, 3473-3481 (2011).</li><li>Werner, P., Blumtritt, H., Zlotnikov, I., Graff, A., Dauphin, Y., Fratzl, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electron+microscope+analyses+of+the+bio-silica+basal+spicule+from+the+Monorhaphis+chuni+sponge.">Electron microscope analyses of the bio-silica basal spicule from the Monorhaphis chuni sponge.</a> J. Struct. Biol. 191 (2), 165-174 (2015).</li><li>Kolednik, O., Predan, J., Fischer, F. 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प्रोटोकॉल के कई कदम है कि बलों और विस्थापन सही मापा जाता है यह सुनिश्चित करने के लिए विशेष रूप से महत्वपूर्ण हैं । जबकि इन महत्वपूर्ण कदम के कुछ सभी तीन सूत्री झुकने परीक्षण के लिए सार्वभौमिक हैं, दूसरो...

<table>
	<tbody>
		<tr>
			<td>TMC 36&#34; x 48&#34; isolation table with 4&#34; CleanTop breadboard</td>
			<td>TMC</td>
			<td>63-563</td>
			<td>Isolation Table</td>
		</tr>
		<tr>
			<td>Diffeential Screw Adjuster</td>
			<td>Thorlabs</td>
			<td>DAS110</td>
			<td>For stage leveling plate</td>
		</tr>
		<tr>
			<td>1&#34; Travel Micrometer Head with 0.001&#34; Graduations</td>
			<td>Thorlabs</td>
			<td>150-801ME</td>
			<td>For stage leveling plate</td>
		</tr>
		<tr>
			<td>Right-Angle Bracket for PT Series Translation Stages, 1/4&#34;-20 Mounting Holes</td>
			<td>Thorlabs</td>
			<td>PT102</td>
			<td>For microscope mount</td>
		</tr>
		<tr>
			<td>1&#34; Dovetail Translation Stage, 1/4&#34;-20 Taps</td>
			<td>Thorlabs</td>
			<td>DT25</td>
			<td>For microscope mount</td>
		</tr>
		<tr>
			<td>1&#34; Translation Stage with 1/4&#34;-170 Adjustment Screw, 1/4&#34;-20 Taps</td>
			<td>Thorlabs</td>
			<td>PT1B</td>
			<td>For microscope mount</td>
		</tr>
		<tr>
			<td>12&#34; Length, Dovetail Optical Rail</td>
			<td>Edmund Optics</td>
			<td>54-401</td>
			<td>For microscope mount</td>
		</tr>
		<tr>
			<td>2.5&#34; Width, Dovetail Carrier</td>
			<td>Edmund Optics</td>
			<td>54-404</td>
			<td>For microscope mount</td>
		</tr>
		<tr>
			<td>0.5&#34; Width, Dovetail Carrier</td>
			<td>Edmund Optics</td>
			<td>54-403</td>
			<td>For microscope mount</td>
		</tr>
		<tr>
			<td>InfiniTube Mounting C-Clamp with &#188;-20</td>
			<td>Edmund Optics</td>
			<td>57-788</td>
			<td>Microscope component</td>
		</tr>
		<tr>
			<td>Standard (with no In-Line Attachment), InfiniTube</td>
			<td>Edmund Optics</td>
			<td>56-125</td>
			<td>Microscope component</td>
		</tr>
		<tr>
			<td>Standard In-Line Attachment (Optimized at 2X-10X), InfiniTube</td>
			<td>Edmund Optics</td>
			<td>56-126</td>
			<td>Microscope component</td>
		</tr>
		<tr>
			<td>Mitutoyo/Achrovid Objective Adapter (M26 to M27)</td>
			<td>Edmund Optics</td>
			<td>53-787</td>
			<td>Microscope component</td>
		</tr>
		<tr>
			<td>5X Infinity Achrovid Microscope Objective</td>
			<td>Edmund Optics</td>
			<td>55-790</td>
			<td>Microscope component</td>
		</tr>
		<tr>
			<td>0.316&#34; ID, Fiber Optic Adapter SX-6</td>
			<td>Edmund Optics</td>
			<td>38-944</td>
			<td>Microscope component</td>
		</tr>
		<tr>
			<td>&#188;&#34; x 36&#34;, Flexible Fiber Optic Light Guide</td>
			<td>Edmund Optics</td>
			<td>42-347</td>
			<td>Microscope component</td>
		</tr>
		<tr>
			<td>115V, MI-150 Fiber Optic Illuminator w/IR Filter and Holder</td>
			<td>Edmund Optics</td>
			<td>55-718</td>
			<td>Microscope component</td>
		</tr>
		<tr>
			<td>Allied Vision Manta G-223 2/3&#34; Color CMOS Camera</td>
			<td>Edmund Optics</td>
			<td>88-452</td>
			<td>Microscope component</td>
		</tr>
		<tr>
			<td>Power Supply for Manta/ Guppy Pro/ Stingray/ Pike</td>
			<td>Edmund Optics</td>
			<td>68-586</td>
			<td>Microscope component</td>
		</tr>
		<tr>
			<td>1/4&#34; Travel Single Axis Translation Stage</td>
			<td>Thorlabs</td>
			<td>MS1S</td>
			<td>FODS micrometer</td>
		</tr>
		<tr>
			<td>Analog Reflectance Dependent Fiber Optic Displacement Sensor</td>
			<td>Philtec</td>
			<td>D20</td>
			<td>FODS</td>
		</tr>
		<tr>
			<td>30V, 3A DC Power Supply</td>
			<td>Agilent</td>
			<td>U8001A</td>
			<td>Power supply for DAQ and FODS</td>
		</tr>
		<tr>
			<td>14-Bit, 48 kS/s Low-Cost Multifunction DAQ</td>
			<td>National Instruments</td>
			<td>USB-6009</td>
			<td>DAQ for FODS</td>
		</tr>
		<tr>
			<td>Three Axis Motorized Translation Stage</td>
			<td>Thorlabs</td>
			<td>Thorlabs T25 XYZ-E/M</td>
			<td>Translation stage</td>
		</tr>
		<tr>
			<td>T-Cube DC Servo Motor Controller</td>
			<td>Thorlabs</td>
			<td>TDC001</td>
			<td>Motor controller for stage</td>
		</tr>
		<tr>
			<td>T-Cube Power Supply</td>
			<td>Thorlabs</td>
			<td>TPS001</td>
			<td>Power supply for motor controller</td>
		</tr>
		<tr>
			<td>National Instruments LabVIEW (2013 SP1)</td>
			<td>National Instruments</td>
			<td></td>
			<td>Used for running software</td>
		</tr>
		<tr>
			<td>National Instruments LabVIEW Vision Acquisition Software (2016)</td>
			<td>National Instruments</td>
			<td></td>
			<td>Used for running software</td>
		</tr>
		<tr>
			<td>Nikon Eclipse Ci-POL Main Body</td>
			<td>MVI</td>
			<td>MDA96000</td>
			<td>Polarized light microscope</td>
		</tr>
		<tr>
			<td>Nikon Pi Intermediate Tube with Analyzer Slider</td>
			<td>MVI</td>
			<td>MDB45305</td>
			<td>Polarized light microscope</td>
		</tr>
		<tr>
			<td>Nikon Dia-Polarizer</td>
			<td>MVI</td>
			<td>MDN11920</td>
			<td>Polarized light microscope</td>
		</tr>
		<tr>
			<td>Power Cord - 7&#39;6&#34;</td>
			<td>MVI</td>
			<td>79035</td>
			<td>Polarized light microscope</td>
		</tr>
		<tr>
			<td>Nikon P-Amh Mechanical Stage</td>
			<td>MVI</td>
			<td>MDC45000</td>
			<td>Polarized light microscope</td>
		</tr>
		<tr>
			<td>Nikon Lwd Achromat Condenser</td>
			<td>MVI</td>
			<td>MBL16100</td>
			<td>Polarized light microscope</td>
		</tr>
		<tr>
			<td>Nikon LV-NBD5BD-CH Manual Quint Nosepiece ESD</td>
			<td>MVI</td>
			<td>MBP60125</td>
			<td>Polarized light microscope</td>
		</tr>
		<tr>
			<td>Nikon C-TF Trinocular Tube F</td>
			<td>MVI</td>
			<td>MBB93100</td>
			<td>Polarized light microscope</td>
		</tr>
		<tr>
			<td>Nikon CFI 10X Eyepiece FN 22mm NC</td>
			<td>MVI</td>
			<td>MAK10110</td>
			<td>Polarized light microscope</td>
		</tr>
		<tr>
			<td>Nikon TU Plan Flour BD 10x Objective</td>
			<td>MVI</td>
			<td>MUE42100</td>
			<td>Polarized light microscope</td>
		</tr>
		<tr>
			<td>Venus Flower Basket Sponge</td>
			<td>Denis Brand</td>
			<td>N/A</td>
			<td>Sponge skeleton</td>
		</tr>
		<tr>
			<td>3.5X Headband Flip-Up Magnifier</td>
			<td>McMaster Carr</td>
			<td>1490T5</td>
			<td>Used for spicule sectioning</td>
		</tr>
		<tr>
			<td>&#216;1&#34; Silicon Wafer, Type P / &#60;100&#62;</td>
			<td>Ted Pella</td>
			<td>16011</td>
			<td>Used for load point mirror</td>
		</tr>
		<tr>
			<td>Low Lint Tapered Tip Cotton Swab</td>
			<td>McMaster Carr</td>
			<td>71035T31</td>
			<td>Used for cleaning LP mirror</td>
		</tr>
		<tr>
			<td>Rubber grip precision knife</td>
			<td>McMaster Carr</td>
			<td>35575A68</td>
			<td>Used for sectioning spicules</td>
		</tr>
		<tr>
			<td>Microscope Slides, frosted end, 75 x 25 x 1mm</td>
			<td>Ted Pella</td>
			<td>260409</td>
			<td>Used for sectioning spicules</td>
		</tr>
		<tr>
			<td>Sable Brushes, #00000, 0.08mm W x 4.0mm L</td>
			<td>Ted Pella</td>
			<td>11806</td>
			<td>Used for handling spicules</td>
		</tr>
		<tr>
			<td>PELCO Pro High Precision Tweezers, extra fine tips, superior finish</td>
			<td>Ted Pella</td>
			<td>5367-5NM</td>
			<td>Used for handling spicules</td>
		</tr>
		<tr>
			<td>Dual Axis Linear Scale Micrometer</td>
			<td>Edmund Optics</td>
			<td>58-608</td>
			<td>Used for calibrating the microscopes</td>
		</tr>
		<tr>
			<td>FLEX-A-TOP FT-38 CAS</td>
			<td>ESD Plastic Containers</td>
			<td>FT-38-CAS</td>
			<td>Used for storing spicules</td>
		</tr>
		<tr>
			<td>Plastic Vial Bullseye Level</td>
			<td>McMaster Carr</td>
			<td>2147A11</td>
			<td>Used for leveling the stage</td>
		</tr>
		<tr>
			<td>Analytical Balance</td>
			<td>Mettler Toledo</td>
			<td>MS105DU</td>
			<td>Used to mass calibration weights</td>
		</tr>
	</tbody>
</table>

a millimeter scale flexural testing system for measuring the mechanical properties of marine sponge spicules

कई लोड असर जैविक संरचनाओं (LBBSs) — जैसे पंख rachises और कंटक — छोटे होते हैं (& #60; 1 mm) लेकिन माइक्रोस्कोपी नहीं । इन LBBSs के वंक व्यवहार को मापने उनके उल्लेखनीय यांत्रिक कार्यों के मूल को समझने के लिए महत्वपूर्ण है ।
हम एक कस्टम निर्मित यांत्रिक परीक्षण डिवाइस है कि 10-5 से 101 एन और विस्थापन 10 से लेकर 10 से लेकर बलों को मापने कर सकते है का उपयोग कर तीन-बिंदु झुका परीक्षण प्रदर्शन के लिए एक प्रोटोकॉल का वर्णन-7 -2 एम । इस यांत्रिक परीक्षण उपकरण का प्राथमिक लाभ यह है कि बल और विस्थापन क्षमता आसानी से अलग LBBSs के लिए समायोजित किया जा सकता है । इस उपकरण का ऑपरेटिंग सिद्धांत एक परमाणु शक्ति माइक्रोस्कोप के समान है । अर्थात्, बल एक लोड बिंदु है कि एक ब्रैकट के अंत से जुड़ा है द्वारा LBBS के लिए लागू किया जाता है । लोड बिंदु विस्थापन एक फाइबर ऑप्टिक विस्थापन संवेदक द्वारा मापा जाता है और एक मापा ब्रैकट कठोरता का उपयोग कर बल में परिवर्तित । डिवाइस के बल रेंज अलग stiffnesses के cantilevers का उपयोग करके समायोजित किया जा सकता है ।
डिवाइस की क्षमताओं समुद्री स्पंज Euplectella aspergillumके कंकाल तत्वों पर तीन सूत्री झुकने परीक्षण प्रदर्शन करके प्रदर्शन कर रहे हैं । कंकाल तत्वों-कंटक के रूप में जाना-सिलिका फाइबर कि लगभग ५० व्यास में µm हैं । हम यांत्रिक परीक्षण उपकरण जांच के लिए प्रक्रियाओं का वर्णन, एक ≈ १.३ mm अवधि के साथ एक तीन-बिंदु झुकने स्थिरता पर कंटक बढ़ते, और एक झुकने परीक्षण प्रदर्शन । लागू बल के स्थान पर spicule और उसके विक्षेपन पर लागू बल मापा जाता है.

किसी भी यांत्रिक परीक्षण के सबसे बुनियादी outputs के नमूने के लिए लागू बल की भयावहता और स्थान है जहां बल लागू किया जाता है पर विस्थापन कर रहे हैं । तीन सूत्री झुकने परीक्षण के मामले में, इस लक्ष्?...

Watch this Scientific Journal Video about A Millimeter Scale Flexural Testing System for Measuring the Mechanical Properties of Marine Sponge Spicules at JoVE.com

A Millimeter Scale Flexural Testing System for Measuring the Mechanical Properties of Marine Sponge Spicules

हम उप मिलीमीटर पैमाने पर एक कस्टम निर्मित यांत्रिक परीक्षण डिवाइस का उपयोग कर फाइबर पर तीन-बिंदु झुकने परीक्षण प्रदर्शन के लिए एक प्रोटोकॉल प्रस्तुत करते हैं । डिवाइस 10 N करने के लिए 20 µN से लेकर बलों उपाय कर सकते हैं और इसलिए फाइबर आकार की एक किस्म को समायोजित कर सकते हैं.

समुद्री स्पंज कंटक के यांत्रिक गुणों को मापने के लिए एक मिलीमीटर पैमाने वंक परीक्षण प्रणाली

Many load bearing biological structures (LBBSs)&#8212;such as feather rachises and spicules&#8212;are small (&#60;1 mm) but not microscopic. Measuring the ...

The overall goal of this experiment is to measure the flexural behavior of fibers whose diameters are between 10 and 100 micrometers. This method can help answer key questions about the mechanical behavior of biological structures, such as the strength and stiffness properties of marine sponge spicules. The main advantage of this technique is that it can be used to measure the mechanical behavior of a wide variety of materials with different sizes and elastic properties.Though this method can provide insight into the mechanical behavior of spicules, it can also be applied to other loading-bearing biological structures, such as plant stems and feather rachises. To begin, attach the load point to the cantilever, using number 4-40 socket head cap screws. Take care not to plastically deform the cantilever arms while attaching the load point.Then, position the load point tip away from the cantilever plate, and loosely attached the cantilever to the plate, using number 6-32 socket head cap screws. Next, insert the 1/8-inch alignment pins through the cantilever and plate, tighten the screws, and then remove the alignment pins. Retract the fiber optic displacement sensor as much as possible by turning the sensor micrometer counterclockwise.Then, loosely attach the cantilever plate to the frame, using number 6-32 socket head cap screws with the load point tip pointing in the negative z-direction. Again, insert the 1/8-inch alignment pins, this time through the frame and cantilever plate, tighten the screws, and then remove the alignment pins. Now, seat the stage on the stage baseplate so that the tips of the micrometer heads on the leveling plate rest in the stage base plate divots.Place a bubble level on the isolation table, and adjust the pressure in each of the table's legs by turning the valve arm thumb screws so that the surface is level. Move the bubble level to the top of the stage leveling plate, and adjust the micrometer so that it is also level. Note the micrometer positions, and remove the stage from the stage baseplate.Use a pair of tweezers to grasp one anchor spicule by its distal end, and pull to remove it from the skeleton. Place the spicule on a clean microscope slide. Using a quintuple zero size red sable brush, hold the spicule against the slide.Cut a four millimeter section of the spicule by pushing a razor blade against the spicule on either side of the brush, perpendicular to the slide surface. Then, discard the distal and proximal spicule sections, and keep the four millimeter section cut from the midpoint. Transfer the spicule section to the sample stage.Position it across the trench with the desired span for the bending test, and gently push it in the positive y-direction against the trench ridge to ensure that the spicule is perpendicular to the trench edges. Seat the stage on the stage baseplate so that the tips of the micrometer spindles rest in the stage baseplate divots. If needed, adjust the micrometers on the stage leveling plate.Open the Bending Test program found in the supplementary code file, and set the Step Size to two micrometers, the Maximum Displacement to 0.5 millimeters, the Low Voltage Stop to 1.5 volts, and the High Voltage Stop to 4.6 volts, using the text boxes shown in the user interface. Select the desired image, data directories, and the output file name, using the text boxes in the user interface. Then, set the Save Images switch in the user interface to the down position, and click the green rectangular button below the words Voltage Difference so that it becomes illuminated.Now, run the Bending Test program, and wait for the motor controller and camera interfaces to initialize. Turn on the illuminator, and adjust the brightness so that the load point tip is visible. Then, turn the fiber optic displacement sensor micrometer clockwise until the output voltage displayed in the user interface graph is about 1.7 volts.Now, use the potentiometer slider on the z-axis motor controller to move the stage in the positive z-direction until it is about one centimeter below the load point tip, and set the z-axis home position by clicking the Home button. Use the potentiometer sliders on the x-and y-axis motor controllers to position the load point tip over the center of the thin steel strip located on the sample stage in the negative x-direction from the trench. Then, use the potentiometer slider on the z-axis motor controller to move the stage in the positive z-direction until the stage is within the microscope's field of view.Click the button labeled Begin Test, and when prompted, enter values of 0.003 volts and 0.001 millimeters for touch sensitivity and touch-off step size, respectively. Click OK, and wait several minutes for the calibration step to finish. Open and run the Basic Data program found in the supplementary code file, and turn the fiber optic displacement sensor micrometer counterclockwise until the output voltage displayed on the user interface graph is approximately three volts.Then, use the potentiometer slider on the x-axis motor controller to position the load point tip between the trench edges above the spicule. Also, use the potentiometer slider on the z-axis motor controller to move the stage in the positive z-direction until the load point tip is below the top surface of the trench ridge. Finally, use the potentiometer slider on the y-axis motor controller to bring the front surface of the trench ridge into focus so that the complete width of the load point tip is between the edges of the trench ridge.Then, stop the Basic Data program by clicking the Stop button. Next, open and run the Center Load Point program, as found in the supplementary code file. Use the x-axis motor controller to move the stage until the load point tip is nearly in contact with the right trench edge.Then, click on the Find Edge button. When prompted, use the x-axis motor controller to move the stage until the load point tip is nearly in contact with the left trench edge. At this point, click on the Find Edge button again, and wait for the program to position the load point tip midway across the trench span.Next, open the Bending Test program. Set the Step Size to two micrometers, Maximum Displacement to 0.5 millimeters, Low Voltage Stop to 1.5 volts, and High Voltage Stop to 4.5 volts using the text boxes in the user interface. Additionally, select the desired image and data directories and the output file name, using the text boxes in the user interface.Set the Save Images switch in the user interface to the up position, and click the green rectangular button below words Voltage Difference so that it is not illuminated. Then, run the Bending Test program, and wait for the motor controller and camera interfaces to initialize. Once initialized, move the stage in the positive z-direction, using the potentiometer slider on the motor controller, until the spicule is within the microscope's field of view.Then, user the potentiometer slider on the y-axis motor controller to move the stage until the spicule is under the load point tip. Next, adjust the microscope focus knob so that the spicule is in focus in the user interface. Then, turn the fiber optic displacement sensor micrometer counterclockwise until the output voltage is approximately 1.8 volts.Once set, click on Begin Test, and wait until the bending test is completed and the stage returns to the z-axis home position. The displacement of the spicule in the z-direction and the force applied by the load point tip can be computed using the voltage-displacement interpolation file, the force calibration file, and the bending test file obtained from the three-point bending test. The voltage-displacement interpolation file is used to measure the cantilever displacement during the bending test.To estimate the stiffness of the cantilever, the force calibration is used, which is then used to relate the cantilever displacement to the force applied by the load point tip. Taken together, these can be used to create the force displacement responses. Shown here, are three different E.aspergillum anchor spicules from successful three-point bending tests.Once mastered, a bending test can be performed with this device in approximately 10 to 15 minutes. The most important aspect of this procedure is ensuring that the spicule is properly seated on the stage and that its axis is perpendicular to the trench edges. Three-point bending tests provide a relatively simple way for researchers studying load-bearing biological structures to gain insight into their mechanical behavior.Following the bending test, beam theories can be used to compute the spicules Young's modulus and fracture strength from the force displacement data.

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