Name: Forfatter Spotlight: Multimodale billeddannelsesstrategier til optimering af lægemiddellevering og tidlig opdagelse i glioblastombehandling
Uploaded: 2024-03-01T14:15:00
Description: Watch this Scientific Journal Video about Two-Photon Intravital Microscopy of Glioblastoma in a Murine Model at JoVE.com

Vi vil gerne takke Stanford Wu Tsai Neuroscience Microscopy Service, Stanford Center for Innovations in Vivo Imaging (SCi3) - small animal imaging center, NIH S10 Shared Instrumentation Grant (S10RR026917-01, PI Michael Moseley, Ph.D.) og Stanford Preclinical Imaging Facility på Porter Drive for at levere udstyr og infrastruktur til dette projekt. Dette arbejde blev støttet af et tilskud fra National Institute for Child Health and Human Development, bevillingsnummer R01HD103638. Vi vil gerne takke Schnitzer-gruppen, Stanford University; Zuo-laboratoriet, Santa Cruz Universitet; og Neurovascular Imaging Laboratory, Boston Photonic Center, University of Boston, til uddannelsesmæssige diskussioner om to-fotonbilleddannelse og kraniale vinduesmodeller.

Den dyreprocedure, der er beskrevet i denne protokol, er i overensstemmelse med kravene fra Det Administrative Panel for Laboratoriedyrs Pleje (APLAC).
1. Cellekultur
<ol><li>Forberedelse af hætte<ol><li>Vask hænder, brug handsker og en laboratoriefrakke. Tænd for det biologiske sikkerhedsskab, og indstil rammeniveauet til en passende åbningshøjde. Lad emhætten rense i 3-5 min. Sprøjt emhætteområdet med 70% ethanol og tør det af med silkepapir.</li>
		<li>Sprøjt a...

Realtidssporing af nanopartikler i hjernetumorer ved hjælp af 2P-IVM i mus

<ol>
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Z., 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=Intubation-free+in+vivo+imaging+of+the+tracheal+mucosa+using+two-photon+microscopy.">Intubation-free in vivo imaging of the tracheal mucosa using two-photon microscopy.</a> Sci Rep. 7 (1), 694 (2017).</li><li>Zong, W., 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=Large-scale+two-photon+calcium+imaging+in+freely+moving+mice.">Large-scale two-photon calcium imaging in freely moving mice.</a> Cell. 185 (7), 1240-1256.e30 (2022).</li><li>Takasaki, K., Abbasi-Asl, R., Waters, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Superficial+bound+of+the+depth+limit+of+two-photon+imaging+in+mouse+brain.">Superficial bound of the depth limit of two-photon imaging in mouse brain.</a> eNeuro. 17 (1), (2019).</li><li>Birkner, A., Konnerth, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deep+two-photon+imaging+in+vivo+with+a+red-shifted+calcium+indicator.">Deep two-photon imaging in vivo with a red-shifted calcium indicator.</a> Methods Mol Biol. 1929, 15-26 (1929).</li><li>Busche, M. 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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Brain+tumor+imaging:+live+imaging+of+glioma+by+two-photon+microscopy.">Brain tumor imaging: live imaging of glioma by two-photon microscopy.</a> Cold Spring Harb Protoc. 2013 (3), (2013).</li><li>Ricard, C., 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=Phenotypic+dynamics+of+microglial+and+monocyte-derived+cells+in+glioblastoma-bearing+mice.">Phenotypic dynamics of microglial and monocyte-derived cells in glioblastoma-bearing mice.</a> Sci Rep. 6, 26381 (2016).</li><li>Soub&#233;ran, A., 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=Effects+of+VEGF+blockade+on+the+dynamics+of+the+inflammatory+landscape+in+glioblastoma-bearing+mice.">Effects of VEGF blockade on the dynamics of the inflammatory landscape in glioblastoma-bearing mice.</a> J Neuroinflammation. 16 (1), 191 (2019).</li><li>Zhang, W., 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=Real-time+vivo+reveals+specific+nanoparticle+target+binding+in+a+syngeneic+glioma+mouse+model.">Real-time vivo reveals specific nanoparticle target binding in a syngeneic glioma mouse model.</a> Nanoscale. 14 (15), 5678-5688 (2022).</li><li>Wartchow, K. M., 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=Treatment+with+cyclic+AMP+activators+reduces+glioblastoma+growth+and+invasion+as+assessed+by+two-photon+microscopy.">Treatment with cyclic AMP activators reduces glioblastoma growth and invasion as assessed by two-photon microscopy.</a> Cells. 10 (3), 556 (2021).</li><li>Jiang, L. W., 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=Label-free+detection+of+fibrillar+collagen+deposition+associated+with+vascular+elements+in+glioblastoma+multiforme+by+using+multiphoton+microscopy.">Label-free detection of fibrillar collagen deposition associated with vascular elements in glioblastoma multiforme by using multiphoton microscopy.</a> J Microsc. 265 (2), 207-213 (2017).</li><li>Chen, Z., Ross, J. 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Vi præsenterer en metode til realtids in vivo NP-sporing ved hjælp af 2P-IVM gennem et kranievindue for at evaluere tumorleveringen af fluorescerende mærkede jernoxid-NP'er. Den kirurgiske teknik til denne procedure kræver en rolig hånd og avancerede eksperimentelle kirurgiske færdigheder. Det er tilrådeligt at øve sig i at bruge slagtekroppe eller fantomer, før du går videre til levende dyreoplevelse. Som et alternativ implementerede Hoeferlin et al. en robotboremaskine for at reducere termisk skade, ...

To-foton intravital mikroskopi (2P-IVM) er en fluorescensbilleddannelsesteknik, der muliggør visualisering af levende væv1.
Først udviklet i 1990'erne er 2P-IVM blevet brugt til in vivo-analyse af nethinden2, nyre3, tyndtarmen4, cochlea5, hjerte6, luftrøret7 og hjernen i forskellige prækliniske modeller 8,9. Inden for neurovidenskab har 2P-IVM fået betydning som en teknik til realtidsbilleddannelse af den sunde hjerne hos vågne dyr10 samt undersøgelse af sygdomme i nervesystemet som Alzheimers11, Parkinsons12 og glioblastom (GBM) 13,14,15,16.
2P-IVM tilbyder en elegant løsning til at studere tumormikromiljøet under udviklingen af GBM. Mens nogle tidligere undersøgelser fokuserede på in vitro17 og ex vivo modeller18, implementerede andre ortopiske19 og xenotropic20 in vivo modeller til undersøgelse af GBM. Madden et al. udførte native imaging af CNS-1 rottegliomcellelinje i en musemodel13. Ved hjælp af en ortopisk GL261-DsRed murinmodel udførte Ricard et al. en intravenøs administration af en fluorofor for at forbedre blodkarrene i tumorområdet i 2P-IVM14.
Her anvender vi 2P-IVM til sporing af tumorlevering af fluorescerende mærkede jernoxidnanopartikler (NP) i en ortopisk musemodel af GBM. Ved hjælp af et kranievindue giver denne metode os mulighed for at studere den rumlige tidsmæssige fordeling af NP'er i realtid i hjernen.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.9% sodium chloride infusion solution</td>
			<td>Baxter Corp</td>
			<td>533-JB1301P</td>
			<td></td>
		</tr>
		<tr>
			<td> 
			Dulbecco&#39;s Modified Eagle Medium</td>
			<td>Invitrogen</td>
			<td>11965-092</td>
			<td></td>
		</tr>
		<tr>
			<td>1 mL syringes</td>
			<td>BD</td>
			<td>Luer-Lok syringe, REF309628</td>
			<td></td>
		</tr>
		<tr>
			<td>10%&#160;FBS</td>
			<td>Thermo fisher</td>
			<td>Cytiva SH30910.03HI</td>
			<td></td>
		</tr>
		<tr>
			<td>10% DMSO</td>
			<td>Sigma-Aldrich</td>
			<td>D8418-50ML</td>
			<td></td>
		</tr>
		<tr>
			<td>2-photon microscope</td>
			<td>Prairie Technologies, Bruker</td>
			<td>Prairie Ultima IV</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol applicators, 70%</td>
			<td>Medline Industries, LP</td>
			<td>MDS093810</td>
			<td></td>
		</tr>
		<tr>
			<td>Alcohol, spray bottle</td>
			<td>Decon Labs Inc</td>
			<td>Decon SaniHol, 04-355-122</td>
			<td></td>
		</tr>
		<tr>
			<td>Aluminum foil</td>
			<td>Reynold Brands</td>
			<td>Reynold Wrap non-stick aluminum foil</td>
			<td></td>
		</tr>
		<tr>
			<td>Anesthesia machine</td>
			<td>Patterson Scientific</td>
			<td>SAS3</td>
			<td></td>
		</tr>
		<tr>
			<td>Anesthesia monitoring&#160;</td>
			<td>Kent Scientific&#160;</td>
			<td>MouseSTAT Jr. Rodent Pulsoxymeter</td>
			<td></td>
		</tr>
		<tr>
			<td>Antibiotic-Antimycotic (100x), liquid</td>
			<td>Invitrogen</td>
			<td>15240-096</td>
			<td></td>
		</tr>
		<tr>
			<td>Betadine applicators</td>
			<td>Professional Disposables International, Inc</td>
			<td>S41125</td>
			<td></td>
		</tr>
		<tr>
			<td>Biopsy punch, 5 mm&#160;</td>
			<td>Miltex</td>
			<td>Size 5</td>
			<td></td>
		</tr>
		<tr>
			<td>Buprenorphine sustained release</td>
			<td>Zoo Pharm</td>
			<td>Bup SR Lab, 1.0 mg/mL</td>
			<td>A generic drug can be used instead.&#160;</td>
		</tr>
		<tr>
			<td>C6 rat glioma cell line</td>
			<td>ATCC (American Tissue Culture Collection)</td>
			<td>CCL-107</td>
			<td></td>
		</tr>
		<tr>
			<td>Cannulas</td>
			<td>BD</td>
			<td>16 G, 1.1/2&#8221;, 30 G, 1&#8221;</td>
			<td></td>
		</tr>
		<tr>
			<td>Carprofen</td>
			<td>Pfizer&#160;</td>
			<td>Rimadyl, 50 mg/ml</td>
			<td>A generic drug can be used instead.&#160;</td>
		</tr>
		<tr>
			<td>Cefazoline</td>
			<td>Sagent Pharmaceuticals</td>
			<td>25021-101-10, 1 g/vial</td>
			<td>A generic drug can be used instead.&#160;</td>
		</tr>
		<tr>
			<td>Cell strainer, 40 &#181;m</td>
			<td>Fisher Scientific</td>
			<td>87711</td>
			<td></td>
		</tr>
		<tr>
			<td>Cotton tip applicators, 6&#8221;&#160;</td>
			<td>Dyad Medical Sourcing, LLC</td>
			<td>HCS1005</td>
			<td></td>
		</tr>
		<tr>
			<td>Dental cement</td>
			<td>Stoelting Co</td>
			<td>51459</td>
			<td>Dental cement kit, clear, 2 components</td>
		</tr>
		<tr>
			<td>Dexamethasone</td>
			<td>Bimeda</td>
			<td>138RX, 2 mg/mL</td>
			<td>A generic drug can be used instead.&#160;</td>
		</tr>
		<tr>
			<td>DietGel</td>
			<td>ClearH2O</td>
			<td>Recovery, 72-06-502</td>
			<td></td>
		</tr>
		<tr>
			<td>Drape&#160;</td>
			<td>Cardinal Health</td>
			<td>Bio Shield Wrap</td>
			<td></td>
		</tr>
		<tr>
			<td>Drill</td>
			<td>Saeyang Microtech</td>
			<td>Escort Pro, B08350</td>
			<td></td>
		</tr>
		<tr>
			<td>Drill tips&#160;</td>
			<td>Hager &#38; Meissinger GmbH</td>
			<td>REF310104001001005</td>
			<td>Size 005, US 1/4</td>
		</tr>
		<tr>
			<td>FIJI imaging analysis software</td>
			<td>National Institute of Health</td>
			<td>https://imagej.net/software/fiji/</td>
			<td></td>
		</tr>
		<tr>
			<td>Forceps</td>
			<td>Fisher Scientific</td>
			<td>13-812-41</td>
			<td></td>
		</tr>
		<tr>
			<td>Gauze</td>
			<td>Fisher HealthCare</td>
			<td>Sterile Cotton Gauze Pad, 4 x 4&#8221;, 22-415-469</td>
			<td></td>
		</tr>
		<tr>
			<td>Gelfoam&#160;</td>
			<td>Ethicon Inc.&#160;</td>
			<td>Surgifoam absorbable gelatin sponge, Ref. 1972</td>
			<td></td>
		</tr>
		<tr>
			<td>Germinator&#160;</td>
			<td>Cellpoint Scientific&#160;</td>
			<td>Germinator 500, No. 11688</td>
			<td></td>
		</tr>
		<tr>
			<td>Glass coverslips, 5 mm diameter</td>
			<td>Fisher Scientific</td>
			<td>Menzel Cover glass&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Gloves, non-sterile</td>
			<td>Fisher Scientific</td>
			<td>Nitrile powder-free medical examination gloves</td>
			<td></td>
		</tr>
		<tr>
			<td>Gloves, sterile</td>
			<td>Medline Industries, LP</td>
			<td>MDS104070</td>
			<td></td>
		</tr>
		<tr>
			<td>Hair removal cream</td>
			<td>Church &#38; Dwight</td>
			<td>Nair Hair remover lotion&#160;</td>
			<td></td>
		</tr>
		<tr>
			<td>Hamilton syringe</td>
			<td>Hamilton Company Inc</td>
			<td>Gastight #1701, 10 &#181;L</td>
			<td></td>
		</tr>
		<tr>
			<td>HBSS without Ca, Mg</td>
			<td>Fisher Scientific</td>
			<td>PI88284</td>
			<td></td>
		</tr>
		<tr>
			<td>Head bar</td>
			<td>Hongway</td>
			<td>5 mm inner diameter O-rings</td>
			<td></td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>Stoelting Co.&#160;</td>
			<td>Rodent warmer X2</td>
			<td></td>
		</tr>
		<tr>
			<td>Insulin syringes</td>
			<td>Exel International Medical Products&#160;</td>
			<td>29G x 1/2&#8243;</td>
			<td></td>
		</tr>
		<tr>
			<td>Iron oxide nanoparticles</td>
			<td>Covis Pharma GmbH</td>
			<td>Feraheme ferumoxytol injection, 510 mg/17 mL, 59338077501</td>
			<td></td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Dechra</td>
			<td>26675-46-7</td>
			<td></td>
		</tr>
		<tr>
			<td>Mice</td>
			<td>Jackson Laboratories</td>
			<td>NSG, Strain 005557</td>
			<td></td>
		</tr>
		<tr>
			<td>Microscope (surgery)</td>
			<td>Seiler Medical</td>
			<td>Seiler IQ Q-100-220</td>
			<td></td>
		</tr>
		<tr>
			<td>Nanoparticles</td>
			<td>Custom</td>
			<td></td>
			<td>Iron oxide nanoparticles (Ferumoxytol) labeled with fluorescein isothiocyanate</td>
		</tr>
		<tr>
			<td>Ophtalmic ointment</td>
			<td>Major pharmaceuticals</td>
			<td>Lubrifresh P.M. nighttime ointment, 203964</td>
			<td></td>
		</tr>
		<tr>
			<td>Oxygen</td>
			<td>Linde Gas &#38; Equipment Inc.&#160;</td>
			<td>High Pressure Steel K Style Cylinder, 249CF, 2000PSIG, CGA 540</td>
			<td></td>
		</tr>
		<tr>
			<td>Plastic cups</td>
			<td>Georgia-Pacirif Consumer Products</td>
			<td>Dixie Portion Cup, 2 oz., Plastic, Clear, PK2400</td>
			<td></td>
		</tr>
		<tr>
			<td>Polyethylene tubing</td>
			<td>Braintree Scientific</td>
			<td>50-195-5494</td>
			<td></td>
		</tr>
		<tr>
			<td>Scale</td>
			<td>Ohaus Corp</td>
			<td>CR2200</td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel</td>
			<td>Integra Life Sciences Production Corp</td>
			<td>Integra Miltex Stainless steel disposable scalpel</td>
			<td></td>
		</tr>
		<tr>
			<td>Scissors</td>
			<td>Fisher Scientific</td>
			<td>13-804-18</td>
			<td></td>
		</tr>
		<tr>
			<td>Sealant</td>
			<td>Henkel Corp</td>
			<td>Loctite 4014</td>
			<td></td>
		</tr>
		<tr>
			<td>Single use lab gown</td>
			<td>High Tech Conversions</td>
			<td>17-444-081</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereotactic frame</td>
			<td>Stoelting Co.&#160;</td>
			<td>Stoelting New Standard TM</td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile Vacuum Bottle Top Filtration Systems</td>
			<td>Fisher Scientific</td>
			<td>S2GPU05RE, MilliporeSigma NO.:S2GPU05RE</td>
			<td></td>
		</tr>
		<tr>
			<td>Styrofoam box</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical gloves</td>
			<td>Cardinal Health</td>
			<td>19-163-108</td>
			<td></td>
		</tr>
		<tr>
			<td>Surgical glue, 3M Vetbond tissues adhesive</td>
			<td>3M Animal Care Prodcuts</td>
			<td>1469SB</td>
			<td></td>
		</tr>
		<tr>
			<td>Tail vein cathether</td>
			<td>Custom</td>
			<td></td>
			<td>Consists of two 30 G cannulas connected with sillicone tubing&#160;</td>
		</tr>
		<tr>
			<td>TrypLE Express (1x), no phenol red</td>
			<td>Invitrogen</td>
			<td>12604-039</td>
			<td></td>
		</tr>
		<tr>
			<td>Ultraviolett torch</td>
			<td>Spring sunshine technology</td>
			<td>Consciot</td>
			<td></td>
		</tr>
	</tbody>
</table>

two-photon intravital microscopy of glioblastoma in a murine model

Leveringen af intravenøst administreret kræftterapi til hjernetumorer er begrænset af blod-hjerne-barrieren. En metode til direkte at afbilde akkumulering og distribution af makromolekyler i hjernetumorer in vivo ville i høj grad forbedre vores evne til at forstå og optimere lægemiddelafgivelse i prækliniske modeller. Denne protokol beskriver en metode til realtids in vivo-sporing af intravenøst administrerede fluorescerende mærkede nanopartikler med to-foton intravital mikroskopi (2P-IVM) i en musemodel af glioblastom (GBM).
Protokollen indeholder en flertrinsbeskrivelse af proceduren, herunder anæstesi og analgesi af forsøgsdyr, oprettelse af et kranievindue, GBM-celleimplantation, placering af en hovedstang, gennemførelse af 2P-IVM-undersøgelser og postkirurgisk pleje til langsigtede opfølgningsundersøgelser. Vi viser repræsentative 2P-IVM-billedbehandlingssessioner og billedanalyse, undersøger fordele og ulemper ved denne teknologi og diskuterer potentielle applikationer.
Denne metode kan let ændres og tilpasses til forskellige forskningsspørgsmål inden for in vivo præklinisk hjernebilleddannelse.

Two-photon intravital microscopy (2P-IVM) is a fluorescence imaging technique that allows the visualization of living tissue1.
First developed in the 1990s, 2P-IVM has been used for in vivo analysis of the retina2, kidney3, small intestine4, cochlea5, heart6, trachea7, and the brain in various preclinical models8,9. In the field of neuroscience, 2P-IVM has gained importance as a technique for real-time imaging of the healthy brain in awake animals10, as well as studying diseases of the nervous system such as Alzheimer&#39;s11, Parkinson&#39;s12 and glioblastoma (GBM)13,14,15,16.
2P-IVM offers an elegant solution for studying the tumor microenvironment during the development of GBM. While some previous studies focused on in vitro17 and ex vivo models18, others implemented orthotopic19 and xenotropic20&#160;in vivo models for examining GBM. Madden et al. performed native imaging of CNS-1 rat glioma cell line in a mouse model13. Using an orthotopic GL261-DsRed murine model, Ricard et al. performed an intravenous administration of a fluorophore to enhance the blood vessels in the tumor region in 2P-IVM14.
Here, we apply 2P-IVM for tracking the tumor delivery of fluorescent-labeled iron oxide nanoparticles (NP) in an orthotopic mouse model of GBM. Using a cranial window, this method allows us to study the real-time spatiotemporal distribution of NPs in the brain in detail.

Forfatter Spotlight: Multimodale billeddannelsesstrategier til optimering af lægemiddellevering og tidlig opdagelse i glioblastombehandling

To-foton intravital mikroskopi af glioblastom i en murin model

Glioblastoma is one of the most common primary brain cancer and is associated with fast growth and poor life expectancy. Developing effective therapies for glioblastoma remains a major challenge since the delivery of therapeutics is limited by the blood brain barrier. A method to directly image the accumulation and distribution of macromolecules in the brain would greatly enhance our ability to understand and optimize drug delivery.Two-photon microscopy offers an elegant solution for studying the real-time distribution of nanoparticles in the preclinical model of glioblastoma. By modifying the nanoparticle used in this study, it's possible to investigate the delivery of different drug candidates and combination therapies on a cellular and molecular level. In addition to two-photon intravital microscopy, the iron oxide component of these nanoparticles allow for multimodal imaging with MRI or MPI.For the future, imaging facilities could serve in the realization of early detection to cancer, thereby leading to more effective therapies against malignant neoplasia. One path to achieve this is to merge two-photon intravital microscopy with other imaging modalities such as positron emission tomography and photoacoustic ultrasound.

Her udførte vi kranievindueskirurgi og indpodede C6-celler i en NSG-musemodel af GBM (n = 5). En korrekt forsegling mellem alle komponenter, der er involveret i oprettelsen af vinduet (figur 1A), vil sikre vinduernes holdbarhed til langvarig billeddannelse og desuden reducere sygeligheden. Ved hjælp af scenen tilpasset in vivo 2P-IVM (figur 2) kunne vi afbilde dyr under anæstesi i op til 2 timer uden større bevægelsesartefakter. Ca. 10 minutter eft...

Watch this Scientific Journal Video about Two-Photon Intravital Microscopy of Glioblastoma in a Murine Model at JoVE.com

Vi præsenterer en ny tilgang til to-fotonmikroskopi af tumorlevering af fluorescerende mærkede jernoxidnanopartikler til glioblastom i en musemodel.

The delivery of intravenously administered cancer therapeutics to brain tumors is limited by the blood-brain barrier. A method to directly image the ...

Glioblastom er en af de mest almindelige primære hjernekræft og er forbundet med hurtig vækst og dårlig forventet levetid. Udvikling af effektive behandlinger for glioblastom er fortsat en stor udfordring, da leveringen af lægemidler er begrænset af blodhjernebarrieren. En metode til direkte at afbilde akkumulering og distribution af makromolekyler i hjernen ville i høj grad forbedre vores evne til at forstå og optimere lægemiddelafgivelse.To-fotonmikroskopi tilbyder en elegant løsning til at studere realtidsfordelingen af nanopartikler i den prækliniske model af glioblastom. Ved at ændre nanopartiklen, der anvendes i denne undersøgelse, er det muligt at undersøge leveringen af forskellige lægemiddelkandidater og kombinationsterapier på cellulært og molekylært niveau. Ud over to-foton intravital mikroskopi muliggør jernoxidkomponenten i disse nanopartikler multimodal billeddannelse med MR eller MPI.For fremtiden kan billeddannelsesfaciliteter tjene til realisering af tidlig påvisning af kræft og derved føre til mere effektive terapier mod ondartet neoplasi. En vej til at opnå dette er at fusionere to-foton intravital mikroskopi med andre billeddannelsesmetoder såsom positronemissionstomografi og fotoakustisk ultralyd.

two-photon-intravital-microscopy-of-glioblastoma-in-a-murine-model

Research

JoVE Journal

Cancer Research

Two-Photon Intravital Microscopy of Glioblastoma in a Murine Model

The delivery of intravenously administered cancer therapeutics to brain tumors is limited by the blood-brain barrier. A method to directly image the accumulation and distribution of macromolecules in brain tumors in vivo would greatly enhance our ability to understand and optimize drug delivery in preclinical models. This protocol describes a method for real-time in vivo tracking of intravenously administered fluorescent-labeled nanoparticles with two-photon intravital microscopy (2P-IVM) in a mouse model of glioblastoma (GBM).
The protocol contains a multi-step description of the procedure, including anesthesia and analgesia of experimental animals, creating a cranial window, GBM cell implantation, placing a head bar, conducting 2P-IVM studies, and post-surgical care for long-term follow-up studies. We show representative 2P-IVM imaging sessions and image analysis, examine the advantages and disadvantages of this technology, and discuss potential applications.
This method can be easily modified and adapted for different research questions in the field of in vivo preclinical brain imaging.

We present a novel approach for two-photon microscopy of the tumor delivery of fluorescent-labeled iron oxide nanoparticles to glioblastoma in a mouse model.