Name: high speed sub-ghz spectrometer for brillouin scattering analysis
Uploaded: 2015-12-22T15:00:00
Description: Watch this Scientific Journal Video about High Speed Sub-GHz Spectrometer for Brillouin Scattering Analysis at JoVE.com

This work was supported in part by the National Institutes of Health (P41-EB015903, R21EY023043, K25EB015885), National Science of Foundation (CBET-0853773) and Human Frontier Science Program (Young Investigator Grant).

Bemærk: Brillouin spektralanalyse kræver et enkelt længde-mode laser (~ 10 mW ved prøven). Til justering øjemed, anvendelse en stærkt svækket del af denne laserstråle (&lt;0,1 mW). 1. Indledende opsætning af Fiber og EMCCD (Electron ganget Charge Coupled Device) Kamera <ol><li> Identificere omkring 1.600 mm fri justering plads til spektrometer på en optisk tabel. </li><li> Monter EMCCD kameraet ved udgangen af ​​den frie justering plads. <ol><li> Brug skruerne til at montere kamerae...

<ol>
	<li>Brillouin, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Diffusion+de+la+lumiere+et+des+rayonnes+X+par+un+corps+transparent+homogene;+influence+del'agitation+thermique.">Diffusion de la lumiere et des rayonnes X par un corps transparent homogene; influence del'agitation thermique.</a> Ann. Phys. (Paris) . 17, 88-122 (1922).</li><li>Scarcelli, G., Yun, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Multistage+VIPA+etalons+for+high-extinction+parallel+Brillouin+spectroscopy.">Multistage VIPA etalons for high-extinction parallel Brillouin spectroscopy.</a> Opt. Exp. 19 (11), 10913-10922 (2011).</li><li>Scarcelli, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Confocal+Brillouin+microscopy+for+three-dimensional+mechanical+imaging.">Confocal Brillouin microscopy for three-dimensional mechanical imaging.</a> Nat. Phot. 2 (1), 39-43 (2008).</li><li>Nichols, A. J., Evans, C. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Video-rate+Scanning+Confocal+Microscopy+and+Microendoscopy.">Video-rate Scanning Confocal Microscopy and Microendoscopy.</a> J. Vis. Exp. (56), e3252 (2011).</li><li>Steelman, Z., Meng, Z., Traverso, A. J., Yakovlev, V. V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brillouin+spectroscopy+as+a+new+method+of+screening+for+increased+CSF+total+protein+during+bacterial+meningitis.">Brillouin spectroscopy as a new method of screening for increased CSF total protein during bacterial meningitis.</a> J. Biophoton. 8 (5), 1-7 (2014).</li><li>Koski, K. J., Yarger, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brillouin+imaging.">Brillouin imaging.</a> Appl. Phys. Lett. 87 (6), 061903 (2005).</li><li>Faris, G. W., Jusinski, L. E., Hickman, A. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=High-resolution+stimulated+Brillouin+gain+spectroscopy+in+glasses+and+crystals.">High-resolution stimulated Brillouin gain spectroscopy in glasses and crystals.</a> J. Opt. Soc. Am. B. 10 (4), 587-599 (1993).</li><li>Scarcelli, G., Yun, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+Brillouin+optical+microscopy+of+the+human+eye.">In vivo Brillouin optical microscopy of the human eye.</a> Opt. Exp. 20 (8), 9197-9202 (2012).</li><li>Scarcelli, G., Besner, S., Pineda, R., Kalout, P., Yun, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+Vivo+Biomechanical+Mapping+of+Normal+and+Keratoconus+Corneas.">In Vivo Biomechanical Mapping of Normal and Keratoconus Corneas.</a> Jama Ophtalmol. , (2015).</li><li>Scarcelli, G., Kim, P., Yun, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=+In+Vivo+Measurement+of+Age-Related+Stiffening+in+the+Crystalline+Lens+by+Brillouin+Optical+Microscopy."> In Vivo Measurement of Age-Related Stiffening in the Crystalline Lens by Brillouin Optical Microscopy.</a> Biophys. J. 101 (6), 1539-1545 (2011).</li><li>Antonacci, G., Foreman, M. R., Paterson, C., T&#246;r&#246;k, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spectral+broadening+in+Brillouin+imaging.">Spectral broadening in Brillouin imaging.</a> Appl. Phys. Lett. 103 (22), 221105 (2013).</li><li>Meng, Z., Traverso, A. J., Yakovlev, V. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Background+clean-up+in+Brillouin+microspectroscopy+of+scattering+medium.">Background clean-up in Brillouin microspectroscopy of scattering medium.</a> Opt. Exp. 22 (5), 5410-5415 (2014).</li><li>Reiss, S., Burau, G., Stachs, O., Guthoff, R., Stolz, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spatially+resolved+Brillouin+spectroscopy+to+determine+the+rheological+properties+of+the+eye+lens.">Spatially resolved Brillouin spectroscopy to determine the rheological properties of the eye lens.</a> Biomed. Opt. Exp. 2 (8), 2144-2159 (2011).</li><li>Scarcelli, G., Kling, S., Quijano, E., Pineda, R., Marcos, S., Yun, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brillouin+microscopy+of+collagen+crosslinking:+noncontact+depth-dependent+analysis+of+corneal+elastic+modulus.">Brillouin microscopy of collagen crosslinking: noncontact depth-dependent analysis of corneal elastic modulus.</a> Invest. Ophthalmol. Vis. Sci. 54 (2), 1418-1425 (2013).</li></ol>

Et centralt træk i denne spektrometer konfiguration er, at de to faser kan justeres uafhængigt af hinanden. Når en VIPA Etalon glides ud af den optiske bane, de resterende linser af spektrometret fase danner en 1: 1 imaging system, således at den spektrale mønster fra hvert trin er afbildet på CCD-kameraet. Derfor er det ligetil at gå tilbage til enten en af ​​de faser for at forbedre sine resultater uden at påvirke tilpasningen af ​​den anden fase. Sættet af translationelle stadier og frihedsgrader for...

Brillouin spredning, først beskrevet af Leon Brillouin 1 i 1922, er uelastisk spredning af lys fra de termiske akustiske modes i et solidt og fra den termiske udsving densitet i en væske eller gas. Den spektrale forskydning af det spredte lys, som regel i sub GHz-området, giver oplysninger om samspillet mellem det indfaldende lys og de akustiske fononer i prøven. Som et resultat, kan det give nyttig information om de viskoelastiske egenskaber af det undersøgte materiale. I sin spontane udgave, generelt Brillouin spredning har tværsnit i størrelsesordenen Raman-spredning, hvilket resulterer i et meget svagt signal. Derudover Brillouin frekvens skift er størrelsesordener mindre end Raman skift. Som følge heraf, elastisk spredte lys (fra Rayleigh eller Mie spredning), falsk lys, og back-off refleksioner af prøven kan alle let overskygge Brillouin spektrale signatur. Derfor, En Brillouin spektrometer skal ikke kun nå sub-GHz spektral opløsning, men også høj spektral kontrast eller udryddelse. I traditionelle Brillouin spektrometre disse krav er opfyldt ved scanning-rivning monokromatorer, optiske bankende metoder og, mest populært, multiple-pass scanning Fabry-Perot interferometre 2. Disse metoder måler hver spektralkomponent sekventielt. Denne tilgang fører til erhvervelse gange for en enkelt Brillouin spektrum lige fra nogle få minutter til flere timer, afhængigt af instrumentet, og på prøven. De to-trins VIPA spektrometer, bygget ved hjælp af denne protokol, har evnen til at samle alle de spektrale komponenter inden for mindre end et sekund samtidig give tilstrækkelig udslettelse (&gt; 60 dB) til effektivt at undertrykke andre falske signaler 2. Integrationen af ​​VIPA etalons er det centrale element i denne spektrometer. En VIPA er en solid etalon med tre forskellige cvariabelt områder: i den forreste overflade, giver en smal antireflekterende belægning strimmel af lys ind i VIPA, mens resten af ​​overfladen har et stærkt reflekterende (HR) belægning; i den bageste overflade, en delvist reflekterende belægning tillader en lille del (~ 5%) af det lys, der skal transmitteres. Når fokuseret på den smalle indgangen til lidt på skrå VIPA, bliver lysstrålen reflekteres i delkomponenter med fast fase forskel i VIPA 2. Interferens mellem sub komponenter opnår den tilstræbte høj spektral spredning. Justering to VIPAs sekventielt på tværs af aksen konfiguration introducerer spektral spredning i vinkelrette retninger 3. Den spektrale dispersion i vinkelrette retninger rumligt adskiller Brillouin toppe fra uønskede krydstale, hvilket gør det muligt kun at stige Brillouin signal. Figur 1 viser en skematisk afbildning af de to trin VIPA spektrometer. Pilene under de optiske elementer angiver degree frihed, hvor translationelle etaper bør være orienteret. <img alt="Figur 1" class="jove_content" p="" src="/files/ftp_upload/53468/53468fig1.jpg" /> Figur 1. Instrumental opsætning. En optisk fiber leverer Brillouin spredning i spektrometer. En cylindrisk linse C1 (f = 200 mm) fokuserer lyset ind i indgangen til den første VIPA (VIPA1). En anden cylindrisk linse C2 (f = 200 mm) kortlægger den spektrale kantede dispersion i en rumlig adskillelse i brændplan C2. I dette plan, er en lodret maske anvendes til at vælge den ønskede del af spektret. Et analogt konfiguration følger, vippes ved 90 grader. Strålen passerer gennem en sfærisk linse S1 (f = 200 mm) og fokuseres i indgangsslidsen for det andet VIPA (VIPA2). En sfærisk linse S2 (f = 200 mm) skaber det todimensionale spektralt adskilte mønster i brændplanet, hvor en anden vandret maske er placeret. Den horsontale maske afbildes på EMCCD kamera ved hjælp af en akromatisk linse par. <a href="https://www.jove.com/files/ftp_upload/53468/53468fig1large.jpg" target="_blank">Klik her for at se en større version af dette tal.</a> En bachelor studerende med nogle optik kursusaktiviteter og grundlæggende justering erfaring bør være i stand til at opbygge og bruge denne to-trins spektrometer. Spektrometeret er for nylig blevet vist at være kompatibel med en række standard optiske prober 3,4,5 (f.eks konfokalt mikroskop, endoskop, spaltelampe oftalmoskop). Her er spektrometer forbundet til et konfokalt mikroskop. Laserlyset er rettet ind i en standard forskning omvendt mikroskop systemet efter at integrere en 90:10 stråledeler. Den tilbagespredning fra prøven kobles til en enkelt mode fiber, hvilket gør mikroskopet konfokal.

<table border="0" cellpadding="0" cellspacing="0" width="573">
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			<td height="21" style="height:21px;width:216px;">OPTICS:</td>
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			<td height="63" style="height:63px;width:216px;">VIPA (virtual image phase array)</td>
			<td style="width:71px;">LIGH MACHINERY</td>
			<td style="width:119px;"></td>
			<td>Quantity: 2</td>
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			<td height="63" style="height:63px;width:216px;">Bundle of Three 423 Linear Stages with SM-25 Micrometers</td>
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			<td style="width:119px;">423-MIC&#160;</td>
			<td>Quantity: 1</td>
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			<td height="42" style="height:42px;">SS Crossed-Roller Bearing Translation Stage, 0.5 in., 8-32, 1/4-20</td>
			<td style="width:71px;">NEWPORT</td>
			<td>9066-X</td>
			<td>Quantity: 1</td>
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			<td height="42" style="height:42px;">Vernier Micrometer, 13 mm Travel, 9 lb Load Capacity, 50.8 TPI</td>
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			<td>SM-13</td>
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			<td height="42" style="height:42px;">Adjustable Width Slit</td>
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			<td>SV-0.5</td>
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			<td height="42" style="height:42px;">Compact Dovetail Linear Stage, 0.20 in. Z Travel, 1.57x1.57x1.38 in.</td>
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			<td>DS40-Z</td>
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			<td height="42" style="height:42px;">Slotted Base Plate, 25 or 40mm to 65mm Stage, 1.1 in. Range</td>
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			<td>B-2B</td>
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			<td height="42" style="height:42px;">&#216;1/2&#34; Optical Post, 8-32 Setscrew, 1/4&#34;-20 Tap, L = 2&#34;, 5 Pack</td>
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			<td>TR2-P5</td>
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			<td height="42" style="height:42px;">&#216;1/2&#34; Post Holders, Spring-Loaded Hex-Locking Thumbscrews, L = 2&#34;, 5 Pack</td>
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			<td>PH2-P5</td>
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			<td height="42" style="height:42px;">&#216;1/2&#34; Post Holders, Spring-Loaded Hex-Locking Thumbscrew, L = 3&#34;, 5 Pack</td>
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			<td>PH3-P5</td>
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			<td height="42" style="height:42px;">Imperial Lens Mount For 2&#34; Optics, 8-32 Tap</td>
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			<td>LMR2</td>
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			<td height="42" style="height:42px;">f=200.0 mm, &#216;2&#34; Achromatic Doublet, ARC: 400-700 nm</td>
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			<td>AC254-200-A</td>
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			<td height="42" style="height:42px;">Kinematic Mount for up to 1.3&#34; (33 mm) Tall Rectangular Optics, Right Handed</td>
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			<td>KM100C</td>
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			<td>CH1A</td>
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			<td height="42" style="height:42px;">f = 200.00 mm, H = 30.00 mm, L = 32.0 mm, N-BK7 Plano-Convex Cylindrical Lens, Antireflection Coating: 350-700 nm</td>
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			<td>L1653L1-A</td>
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			<td>RA90</td>
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			<td>SM1A9</td>
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			<td>PB4</td>
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			<td height="42" style="height:42px;">Spacer, 2&#34; x 3&#34;, 1.000&#34; Thick</td>
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			<td height="42" style="height:42px;">543 nm, f=15.01 mm, NA=0.17 FC/APC Fiber Collimation Pkg.</td>
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			<td>F260APC-A</td>
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			<td height="42" style="height:42px;">SM1-Threaded Adapter for &#216;11 mm collimators</td>
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			<td>Ad11F</td>
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			<td height="42" style="height:42px;">Translating Lens Mount for &#216;1&#34; Optics, 1 Retaining Ring Included</td>
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			<td>LM1XY</td>
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			<td height="42" style="height:42px;">Single Mode Patch Cable, 450 - 600 nm, FC/APC, 2 m Long</td>
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			<td>P3-460B-FC-2</td>
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			<td height="42" style="height:42px;">1:1 Matched Achr. Pair, f1=30 mm, f2=30 mm, BBAR 400-700 nm</td>
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			<td>MAP103030-A</td>
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			<td height="42" style="height:42px;">SM1 Lens Tube&#8230;length to adjust depend on CCD, we have 3.5 inches</td>
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			<td>SM1LXX</td>
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			<td height="42" style="height:42px;">25 mm Construction Rail, L = 15&#34;</td>
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			<td height="42" style="height:42px;">High Performance Black Masking Tape, 2&#34; x 60 yds. (50 mm x 55 m) Roll</td>
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			<td>T743-2.0</td>
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			<td>XE25T3</td>
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			<td height="42" style="height:42px;">&#160;1/4&#34;-20 Low-Profile Channel Screws (100 Screws/Box)</td>
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			<td>SH25LP38</td>
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			<td height="42" style="height:42px;">60&#34; (W) x 3 yds. (L) x 0.005&#34; (T) (1.5 m x 2.7 m x 0.12 mm) Blackout Fabric</td>
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high speed sub-ghz spectrometer for brillouin scattering analysis

Målet med denne protokol er at opbygge en parallel høj udslettelse og høj opløsning optisk Brillouin spektrometer. Brillouin spektroskopi er en berøringsfri målemetode, der kan anvendes til at opnå direkte udlæsninger af viskoelastisk materialeegenskaber. Det har været et nyttigt redskab i materiale karakterisering, strukturel overvågning og miljømæssig sensing. Tidligere har Brillouin spektroskopi sædvanligvis scanning Fabry-Perot etalons at udføre spektralanalyse. Denne proces kræver høje belysning effekt og lange erhvervelsestider, hvilket gør teknikken uegnet til biomedicinske anvendelser. En nylig indført nye spektrometer overvinder denne udfordring ved at ansætte to VIPAs på tværs af aksen konfiguration. Denne nyskabelse gør det muligt for sub-gigahertz (GHz) opløsning spektralanalyse med sub-anden erhvervelse tid og belysning magt inden de sikkerhedsmæssige grænser for biologisk væv. De mange nye applikationer lettes af denne forbedring er currently blive undersøgt i biologisk forskning og klinisk anvendelse.

<html> Figur 3 viser repræsentative Brillouin spektre og deres passer til forskellige materialer. De VIPAs begge har en tykkelse på 5 mm, hvilket resulterer i en FSR på ca. 20 GHz. Integrationstiden for disse målinger var 100 msek. 100 målinger blev taget og gennemsnit. Én kalibrering måling blev taget før erhverve spektre. <img alt="Figur 3" src="/files/ftp_upload/53468/53468fig3.jpg" /> Figur 3. Brillouin spektre af...

Watch this Scientific Journal Video about High Speed Sub-GHz Spectrometer for Brillouin Scattering Analysis at JoVE.com

High Speed Sub-GHz Spectrometer for Brillouin Scattering Analysis

Her præsenterer vi en protokol til at bygge en hurtig Brillouin spektrometer. Cascading næsten afbildet fase array (VIPA) etalons opnå en måling hastighed mere end 1.000 gange hurtigere end traditionel scanning Fabry-Perot spektrometre. Denne forbedring giver midlerne til Brillouin analyse af væv og biomaterialer ved lave effektniveauer in vivo.

High Speed ​​Sub-GHz spektrometer til Brillouin spredning Analyse

The goal of this protocol is to build a parallel high-extinction and high-resolution optical Brillouin spectrometer. Brillouin spectroscopy is a ...

Research

JoVE Journal

Bioengineering

Polymer Microarrays for High Throughput Discovery of Biomaterials

The discovery of novel biomaterials that are optimized for a specific biological application is readily achieved using polymer microarrays, which allows a ...

Evaluation of Polymeric Gene Delivery Nanoparticles by Nanoparticle Tracking Analysis and High-throughput Flow Cytometry

Evaluering af Polymeric Gene Delivery nanopartikler ved Nanopartikel Tracking Analyse og Højkapacitetsforskning Flowcytometri

Non-viral gene delivery using polymeric nanoparticles has emerged as an attractive approach for gene therapy to treat genetic diseases1 and as a ...

Self-reporting Scaffolds for 3-Dimensional Cell Culture

Selvrapportering Stilladser for 3-Dimensional Cell Culture

Culturing cells in 3D on appropriate scaffolds is thought to better mimic the in vivo microenvironment and increase cell-cell interactions. The resulting ...

High-resolution Spatiotemporal Analysis of Receptor Dynamics by Single-molecule Fluorescence Microscopy

Høj opløsning Spatiotemporal Analyse af receptor Dynamics ved Single-molekyle fluorescensmikroskopi

Single-molecule microscopy is emerging as a powerful approach to analyze the behavior of signaling molecules, in particular concerning those aspect (e.g., ...

A Guide to Structured Illumination TIRF Microscopy at High Speed with Multiple Colors

En guide til Structured Illumination TIRF Mikroskopi på High Speed ​​med flere farver

Optical super-resolution imaging with structured illumination microscopy (SIM) is a key technology for the visualization of processes at the molecular ...

Preparation of Extracellular Matrix Protein Fibers for Brillouin Spectroscopy

Fremstilling af ekstracellulært matrixprotein fibre til Brillouin Spektroskopi

Brillouin spectroscopy is an emerging technique in the biomedical field. It probes the mechanical properties of a sample through the interaction of ...

A High-throughput Cell Microarray Platform for Correlative Analysis of Cell Differentiation and Traction Forces

En High-throughput Cell Microarray Platform for korrelative Analyse af Cell Differentiering og Traction Forces

Microfabricated cellular microarrays, which consist of contact-printed combinations of biomolecules on an elastic hydrogel surface, provide a tightly ...

Implementation of Interference Reflection Microscopy for Label-free, High-speed Imaging of Microtubules

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There are several methods for visualizing purified biomolecules near surfaces. Total-internal reflection fluorescence (TIRF) microscopy is a commonly used ...

Imaging and Quantification of the Area of Fast-Moving Microbubbles Using a High-Speed Camera and Image Analysis

Billeddannelse og kvantificering af området for hurtige mikrobobler ved hjælp af et højhastighedskamera og billedanalyse

An experimental and image analysis technique is presented for imaging cavitation bubbles and calculating their area. The high-speed imaging experimental ...

High Speed ​​Sub-GHz spektrometer til Brillouin spredning Analyse

High Speed Sub-GHz spektrometer til Brillouin spredning Analyse