Name: a bright nir-ii fluorescence probe for vascular and tumor imaging
Uploaded: 2023-03-17T14:55:00
Description: Watch this Scientific Journal Video about A Bright NIR-II Fluorescence Probe for Vascular and Tumor Imaging at JoVE.com

Dette arbejde blev delvist støttet af tilskud fra NSFC (82273796, 82111530209), Special Funds for Guiding Local Science and Technology Development of Central Government (XZ202202YD0021C, XZ202102YD0033C, XZ202001YD0028C), Hubei-provinsens videnskabelige og tekniske innovationsnøgleprojekt (2020BAB058), grundforskningsmidlerne til de centrale universiteter og Tibet autonome region COVID-19 forebyggelses- og kontrolprogrammer for videnskab og teknologiudvikling.

Dyreforsøg til NIR-II-billeddannelsesundersøgelser blev udført på Animal Experiment Center ved Wuhan University, som er blevet tildelt International Association for Experimental Animal Care (AALAC). Alle dyreforsøg blev udført i henhold til China Animal Welfare Commission Guidelines for the Care and Use of Experimental Animals og godkendt af Animal Care and Use Committee (IACUC) fra Animal Experimental Center ved Wuhan University.
Hunmus med BALB/c nøgen mus (~20 g) ved 6 ugers alderen ...

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NIR-I fluorescerende billeddannelse kan til en vis grad anvendes til tumor- og vaskulær billeddannelse, men på grund af den begrænsede maksimale emissionsbølgelængde af NIR-I-fluoroforer (<900 nm) resulterer det i dårlig vævspenetration og tumorsignalbaggrundsforhold33,34. Dårlig og lav billedopløsning kan forårsage en afvigelse mellem resultatet af billedfeedbackbehandlingen og den faktiske terapeutiske effekt. Derudover har de fleste NIR-I-fluoroforer...

Fluorescensbilleddannelse er det almindeligt anvendte molekylære billeddannelsesværktøj i grundforskning og bruges også ofte til at guide kirurgisk tumorresektion i klinikker1. Det grundlæggende princip for fluorescensbilleddannelse er at anvende et kamera til at modtage fluorescens udsendt af en laser efter bestråling af prøver (væv, organer osv.) 2. Processen afsluttes inden for få millisekunder3. Fluorescensbilleddannelsesbølgelængderne kan opdeles i ultraviolet (200-400 nm), synligt område (400-700 nm), nær-infrarød I (NIR-I, 700-900 nm) og nær-infrarød II (NIR-II, 1000-1700 nm)4,5,6. Fordi de endogene molekyler såsom hæmoglobin, melanin, deoxyhemoglobin og bilirubin i biologiske væv har stærk absorption og en spredningseffekt på lyset i synlige områder, reduceres lysets penetration og følsomhed kraftigt, og fluorescensbilleddannelse i synlige lysbølgelængder påvirkes negativt 7,8,9.
NIR-II fluorescensbilleddannelse har lav fotonabsorption og spredning, høj billedhastighed og høj billedkontrast (eller følsomhed)10,11. Når fluorescensbølgelængden øges, falder absorptionen og spredningen af fluorescens i biologiske væv gradvist, og autofluorescensen i NIR-II-regionen er ekstremt lav12. Således øger NIR-II-vinduet signifikant penetrationsdybden af væv og opnår en højere opløsning og signal-støj-forhold13,14,15. NIR-II-vinduet kan yderligere opdeles i NIR-IIa (1300-1400 nm) og NIR-lIb (1500-1700 nm) vinduer16. Til dato er der rapporteret flere milepæle NIR-II-materialer, herunder uorganisk materiale enkeltvæggede kulstofnanorør, sjældne jordarters nanopartikler, kvantepunkter og organiske materiale halvlederpolymernanopartikler, småmolekylefarvestoffer, aggregeringsinducerede selvlysende materialer osv. 1,17,18,19,20,21,22. Uorganiske nanomaterialer akkumuleres let i leveren, milten osv. og har potentiel biotoksicitet på lang sigt23. Organisk småmolekylefluorofor har fordelene ved hurtig metabolisme, lav toksicitet, let modifikation og en klar struktur, som er den mest lovende sonde til klinisk brug24.
NIR-II-billeddannelsessystemet er også en kritisk komponent i fluorescensbiobilleddannelse, fordi det effektivt kan indsamle NIR-II-fluorescenssignaler fra NIR-II-sonden og dermed gengive præcise funktionelle, anatomiske og molekylære billeder25,26. NIR-II-billedbehandlingssystemet består hovedsageligt af kortbølgede infrarøde kameraer, langpasfiltre (LP), lasere og computerprocessorer. In vivo NIR-II fluorescerende billeddannelse betragtes som en af de mest gennemførlige billeddannelsesmetoder til belysning af sygdomsmekanismer og livets natur27,28,29. NIR-II-billeddannelsesteknologi er blevet brugt i vid udstrækning inden for biomedicinske områder såsom kræftcelledetektion, dynamisk billeddannelse, in vivo målrettet sporing og målrettet terapi, især inden for onkologisk forskning30,31. I betragtning af de høje tekniske krav til NIR-II-billeddannelsesteknologi på billedsonder og instrumenter pusler og begrænser det også den praktiske anvendelse af forskere på forskellige områder. Derfor introduceres forberedelsen af NIR-II-billeddannelsessonder og anvendelserne af NIR-II-billeddannelse detaljeret i denne artikel.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Anhydrous pyridine</td>
			<td>Perimed&#160;</td>
			<td>110-86-1</td>
			<td></td>
		</tr>
		<tr>
			<td>Anhydrous sodium sulfate</td>
			<td>China national medicines Co.,Ltd</td>
			<td>SY006376</td>
			<td></td>
		</tr>
		<tr>
			<td>Black cardboard</td>
			<td>Suzhou Yingrui Optical Technology Co., Ltd</td>
			<td>AO00158</td>
			<td></td>
		</tr>
		<tr>
			<td>Column chromatography</td>
			<td>Energy Chemical</td>
			<td>E080498</td>
			<td></td>
		</tr>
		<tr>
			<td>Diphenylphosphine palladium dichloride</td>
			<td>Sigma-Aldrich</td>
			<td>B2161-1g</td>
			<td></td>
		</tr>
		<tr>
			<td>DSPE-PEG2000</td>
			<td>Ponsure</td>
			<td>PS-E1</td>
			<td></td>
		</tr>
		<tr>
			<td>Dulbecco&#39;s modified eagle medium&#160;</td>
			<td>Gibco</td>
			<td>8121587</td>
			<td></td>
		</tr>
		<tr>
			<td>EGTA</td>
			<td>Biofroxx</td>
			<td>EZ6789D115</td>
			<td></td>
		</tr>
		<tr>
			<td>Fetal bovine serum</td>
			<td>Gibco</td>
			<td>2166090RP</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>GLPBIO</td>
			<td>GC45487-1</td>
			<td></td>
		</tr>
		<tr>
			<td>K2CO3</td>
			<td>Macklin</td>
			<td>P816305-5g</td>
			<td></td>
		</tr>
		<tr>
			<td>N. N &#39;- dimethylformamide</td>
			<td>China national medicines Co.,Ltd</td>
			<td>02-12-1968</td>
			<td></td>
		</tr>
		<tr>
			<td>NIR-II imaging instrument</td>
			<td>Suzhou Yingrui Optical Technology Co., Ltd</td>
			<td>16011109</td>
			<td></td>
		</tr>
		<tr>
			<td>N-sulfenanilide</td>
			<td>Enerry chemical&#160;</td>
			<td>1250030-5g</td>
			<td></td>
		</tr>
		<tr>
			<td>PdCl2(dppf)2CH2Cl2</td>
			<td>TCI&#160;</td>
			<td>B2064-1g</td>
			<td></td>
		</tr>
		<tr>
			<td>penicillin-streptomycin</td>
			<td>Gibco</td>
			<td>15140-122</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetrahydrofuran</td>
			<td>China national medicines Co.,Ltd</td>
			<td>M005197</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetratriphenylphosphine palladium</td>
			<td>Immochem</td>
			<td>1021232-5g</td>
			<td></td>
		</tr>
		<tr>
			<td>Tetratriphenylphosphine palladium</td>
			<td>Sigma-Aldrich</td>
			<td>1021232-5g</td>
			<td></td>
		</tr>
		<tr>
			<td>Tributyltin chloride</td>
			<td>Immochem</td>
			<td>QH004335</td>
			<td></td>
		</tr>
		<tr>
			<td>Trimethylchlorosilane</td>
			<td>China national medicines Co.,Ltd</td>
			<td>40060560</td>
			<td></td>
		</tr>
	</tbody>
</table>

a bright nir-ii fluorescence probe for vascular and tumor imaging

Som en ny billeddannelsesteknologi har nær-infrarød II (NIR-II, 1000-1700 nm) fluorescensbilleddannelse et betydeligt potentiale på det biomedicinske område på grund af dets høje følsomhed, dybe vævsindtrængning og overlegen billeddannelse med rumlig og tidsmæssig opløsning. Metoden til at lette implementeringen af NIR-II-fluorescensbilleddannelse til nogle presserende nødvendige områder, såsom medicinsk videnskab og farmaci, har imidlertid undret relevante forskere. Denne protokol beskriver detaljeret konstruktionen og bioimaging anvendelser af en NIR-II fluorescens molekylær sonde, HLY1, med et D-A-D (donor-acceptor-donor) skelet. HLY1 viste gode optiske egenskaber og biokompatibilitet. Desuden blev NIR-II vaskulær og tumorbilleddannelse hos mus udført ved hjælp af en NIR-II optisk billeddannelsesenhed. NIR-II-fluorescensbilleder i realtid i høj opløsning blev erhvervet for at guide påvisning af tumorer og vaskulære sygdomme. Fra sondeforberedelse til dataindsamling forbedres billedkvaliteten betydeligt, og ægtheden af NIR-II-molekylære sonder til dataregistrering i intravital billeddannelse sikres.

Den fluorescerende intensitet og lysstyrke af vandspændende HLY1-prikker blev bestemt af et NIR-II-billeddannelsesinstrument. Den fluorescerende intensitet af HLY1 i 90% fw THF/H2O-blandingenvar fem gange så høj som i THF-opløsningen, hvilket indikerede et fremtrædende AIE-træk ved HLY1 (figur 1B). Desuden udsendte HLY1-prikker stærke fluorescerende signaler under et 1.500 nm LP-filter, hvilket viser, at HLY1-prikker kan bruges til NIR-IIb-billeddannelse...

Watch this Scientific Journal Video about A Bright NIR-II Fluorescence Probe for Vascular and Tumor Imaging at JoVE.com

A Bright NIR-II Fluorescence Probe for Vascular and Tumor Imaging

Denne protokol beskriver en detaljeret, real-time NIR-II fluorescens imaging operation af en mus ved hjælp af en NIR-II optik billeddannelse enhed.

En lys NIR-II fluorescenssonde til vaskulær og tumorbilleddannelse

As an emerging imaging technology, near-infrared II (NIR-II, 1000-1700 nm) fluorescence imaging has significant potential in the biomedical field, owing ...

Højopløselig vaskulær og tumorbilleddannelse blev realiseret af den nær-infrarøde til fluorescerende nanoprob, hvilket giver en nøjagtig og effektiv metode til styring af vaskulære sygdomme og kræft. In vivo-billeddannelse i NIR-II-vinduet, som gør det muligt for os at se dybt ind i levende emner, skaber muligheder for biomedicinsk forskning og vores kliniske anvendelser. Forbered dig på nær-infrarød II- eller NIR-II-billeddannelse ved at placere kommercielt tilgængeligt sort pap i midten af billeddannelsesenhedens bærer.Placer derefter prøven oven på det sorte pap, så det er i midten af bæreren. Når du har valgt det relevante filter, skal du udføre platformshøjdejusteringen. Tryk længe på muligheden for platform op på berøringsskærmgrænsefladen i bærekonsollens kontrolområde for at flytte den opad.Og tryk platformen længe ned for at flytte den nedad. Efter syntetisering af NIR-II-farvestoffet HLY1 fremstilles HLY1-prikkerne ved nanoudfældningsmetode ved anvendelse af DSPE-PEG-2K som indkapslingsmatrix. For at gøre det skal du placere et bægerglas indeholdende en opløsning af 10 milligram DSPE-PEG-2K i ni ml vand til sonikering ved 25 grader Celsius.Derefter opløses et milligram HLY1 i en milliliter tetrahydrofuran eller THF. Og tilsæt langsomt opløsningen i DSPE-PEG-2K vandig opløsning, der gennemgår sonikering. Fjern derefter THF fra blandingen ved dialyse.Efter koncentrering af ovennævnte opløsning centrifugalt med ultrafiltrering ved 7.100 G i 10 minutter, anbringes den i et køleskab med fire grader Celsius til fremtidig brug. Til in vivo-billeddannelse injiceres HLY1-prikkerne intravenøst i de bedøvede mus. Og tre minutter senere, fortsæt med at udføre NIR-II fluorescensbilleddannelse af blodkarrene i hele musens krop ved hjælp af et optisk NIR-II-billeddannelsessystem.Fokuser yderligere på musens hoved for at indsamle hjernevaskulær billeddannelse. Indstil instrumentparametrene for det optiske NIR-II-billeddannelsessystem til 90 milliwatt pr. Kvadratcentimeter og 808 nanometer laser. Indsaml billederne fem minutter efter injektionen af HLY1-prikker i mus, og behandl dataene ved hjælp af ImageJ-software.Fluorescensintensiteten af HLY1 i 90%THF-opløsningen indeholdende 90% vand var fem gange så høj som i THF-opløsningen, hvilket indikerer en fremtrædende aggregeringsinduceret emission eller AIE-funktion af HLY1. HLY1-prikkerne udsendte stærke fluorescerende signaler under et 1.500 nanometer lavpas- eller LP-filter, der fastslog dets egnethed til NIR-IIb-billeddannelse. Den maksimale absorptions- og emissionsbølgelængde af HLY1-prikker var henholdsvis 740 nanometer og 1.040 nanometer.Derudover blev den hydrodynamiske størrelse af HLY1-prikker bestemt til at være 145 nanometer ved dynamisk lysspredning. Hos mus administreret med HLY1-prikker blev cerebrale kar og mikrokarrene i bagbenet tydeligt identificeret ved NIR-II-billeddannelse under et 1.500 nanometer LP-filter. Endvidere var de fire T1-tumorer hos de tumorbærende mus også tydeligt synlige ved NIR-II-billeddannelse, hvilket indikerer den forbedrede permeabilitet og retention eller EPR-effekt af HLY1-prikker.Alle disse resultater antydede, at HLY1-prikker er en lys NIR-II-fluorescenssonde, der kan anvendes til vaskulær og tumorbilleddannelse. Forberedelse af den vandsuspenderbare nanosonde under in vivo- og NIR-II-fluorescensbilleddannelse er den vigtigste del af proceduren.

Research

JoVE Journal

Medicine

In vivo Near Infrared Fluorescence (NIRF) Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

In vivo Near Infrared Fluorescence NIRF Intravascular Molecular Imaging of Inflammatory Plaque, a Multimodal Approach to Imaging of Atherosclerosis

The vascular response to injury is a well-orchestrated inflammatory response triggered by the accumulation of macrophages within the vessel wall leading ...

Monitoring Tumor Metastases and Osteolytic Lesions with Bioluminescence and Micro CT Imaging

Following intracardiac delivery of MDA-MB-231-luc-D3H2LN cells to Nu/Nu mice, systemic metastases developed in the injected animals. Bioluminescence ...

Multi-photon Imaging of Tumor Cell Invasion in an Orthotopic Mouse Model of Oral Squamous Cell Carcinoma

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

In vivo Imaging of Tumor Angiogenesis using Fluorescence Confocal Videomicroscopy

In vivo Imaging of Tumor Angiogenesis using Fluorescence Confocal Videomicroscopy

In vivo billeddannelse af tumorangiogenese hjælp Fluorescens Confocal Videomicroscopy

Fibered confocal fluorescence in vivo imaging with a fiber optic bundle uses the same principle as fluorescent confocal microscopy. It can excite ...

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

A Novel High-resolution In vivo Imaging Technique to Study the Dynamic Response of Intracranial Structures to Tumor Growth and Therapeutics

A Novel Høj opløsning<em&gt; In vivo</em&gt; Imaging teknik til at undersøge den dynamiske respons af Intrakranielle Structures til tumorvækst og Therapeutics

We have successfully integrated previously established Intracranial window (ICW) technology 1-4 with intravital 2-photon confocal microscopy to develop a ...

Osmotic Drug Delivery to Ischemic Hindlimbs and Perfusion of Vasculature with Microfil for Micro-Computed Tomography Imaging

Osmotisk Drug Delivery til Iskæmisk bagbensknogler og perfusion af kar med Microfil for Micro-computertomografi Imaging

Preclinical research in animal models of peripheral arterial disease plays a vital role in testing the efficacy of therapeutic agents designed to ...

Computerized Dynamic Posturography for Postural Control Assessment in Patients with Intermittent Claudication

Computerized dynamic posturography with the EquiTest is an objective technique for measuring postural strategies under challenging static and dynamic ...

Evaluation of Tumor-infiltrating Leukocyte Subsets in a Subcutaneous Tumor Model

Evaluering af tumorinfiltrerende leukocytundergrupper i en subkutan Tumor Model

Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or ...

Establishment of a Human Multiple Myeloma Xenograft Model in the Chicken to Study Tumor Growth, Invasion and Angiogenesis

Etablering af en human myelomatose xenograftmodel i kylling til Studieinformationen tumorvækst, invasion og angiogenese

Multiple myeloma (MM), a malignant plasma cell disease, remains incurable and novel drugs are required to improve the prognosis of patients. Due to the ...

Visualization of Vascular and Parenchymal Regeneration after 70% Partial Hepatectomy in Normal Mice

Visualisering af Vascular og parenkymale Regeneration efter 70% Delvis hepatektomi i normale mus

A modified silicone injection procedure was used for visualization of the hepatic vascular tree. This procedure consisted of in-vivo injection of the ...