Name: in vivo imaging of cx3cr1gfp/gfp reporter mice with spectral-domain optical coherence tomography and scanning laser ophthalmoscopy
Uploaded: 2017-11-11T14:00:00
Description: Watch this Scientific Journal Video about In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy at JoVE.com

Bu eser İsviçre Ulusal Bilim Vakfı (SNSF; #320030_156019) bir grant tarafından desteklenmiştir. Yazarlar Heidelberg Engineering GmBH, Almanya kelimelerinde destek aldı.

tüm yordamlarda kullanılan 24 kim gfp Cx3cr1 organizatörü altında hızlı BALB/c yetişkin erkek ve dişi fareler vardı. Fareler ARVO açıklamaya göre Ophthalmic ve vizyon araştırma hayvanlar kullanımı için tedavi edildi ve tüm yordamları hayvan refah İsviçre Federal düzenlemelerini göre İsviçre hükümetinin kabul edildi. Fareler medetomidine hidroklorid (0.75 mg/kg) ve ketamin (45 mg/kg) subkutan enjeksiyon tarafından anestezi. Uygun anestezi olduğunu doğrulad?...

<ol>
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Mevcut madde retina B-inceden inceye gözden geçirmek toplama ve görüntüleme gfp olumlu microglia dağıtım fare retina görüntüleme aynı oturumda bir iletişim kuralı gösterir. SD-Ekim ve SLO giderek retina hastalığı hayvan modellerinde saat10,14,17,18,21üzerinde retina değişikliklerini bilgi sağlamak için kullanılır. Bu iletişim kuralı,...

Deneysel Oftalmoloji retina patoloji incelenmesi genellikle histolojik teknikler kullanılarak değerlendirilir. Ancak, Histoloji hayvan euthanization gerektirir ve değişiklik doku güncel özellikler için neden olabilir. SD-Ekim ve SLO rutin klinik Oftalmoloji tanı amaçlı ve diyabetik makula ödemi9, anterior iskemik optik nöropati10veya retinitis pigmentosa11 gibi birkaç retina hastalıkları izleme kullanılır . SD-Ekim ve SLO daha fazla müdahale olmadan dilate pupil aracılığıyla görüntülenmiştir retina, yüksek çözünürlüklü görüntüler oluşturmak non-invaziv teknikler vardır. SD-Ekim SLO retina stereoskopik yüksek kontrastlı görüntüler üretmek için Floresans veri toplarken retina, kesitsel görüntüler oluşturmak için gelmekte veri toplayarak retina yapısı ve retina kalınlık bilgi sağlar. Günümüzde, her iki tekniğin de giderek daha küçük kemirgen12,13,14,15 (ya da bile Zebra balığı16,17) kullanarak deneysel Oftalmoloji kullanılır ve olabilir hem nitel ve nicel bilgi12,17,18,19,20,21sağlar.
Lipofuscins gibi endojen fluorophores birikimi ya da retina drusen oluşumu bu SLO tarafından auto floresan sinyali olarak görüntülenmeyecektir. Bu özellik, tanı ve yaşa bağlı makula dejenerasyonu veya retinitis pigmentosa22,23gibi retina hastalıkları izleme için değerli bir tekniği SLO yapar. Deneysel Oftalmoloji içinde görüntüleme (AF) otomatik Floresans belirli hücre tipleri muhabir fare satırlarındaki tespiti için kullanılabilir. Örneğin, fare heterozigoz gfp Cx3cr124 organizatörü altında bir ifade için in vivo görselleştirme mikroglial hücrelerinin normal retina ve microglia/makrofaj incelenmesi için avantajlı olan Dynamics retina hastalığı21. Microglia doku homeostazı ve doku onarımı yaralanma1,25,26üzerine çok önemli bir rol oynamak retina, ikamet makrofajlar vardır. Microglia harekete geçirmek içinde retina retina yaralanma, iskemi ve dejenerasyonu retina hastalığı2,3,4,5, bu hücrelerinde bir rolü düşündüren, bildirilmiştir 6.
Amacı mevcut protokolü, retina görüntüleme ve SD-Ekim kullanarak ve retina kalınlık ölçüm yapılmasına ve görselleştirme Cx3cr1gfp/gfp fare retina kullanarak gfp olumlu microglia hücreler için nispeten basit bir yöntem tarif etmektir SLO (Heidelberg Spectralis HRA + OCT sistemi). Bu iletişim kuralı çeşitli fare satırlarındaki sağlıklı ya da hasta retina görüntüleme ve kalınlığı ölçümleri için yararlı olabilir. Ayrıca, xarakteristikaları analizleri tanıma ve miktar microglia sayı ve microglia harekete geçirmek SLO21kullanarak retina içinde gerçekleştirilir. Merkezi sinir sistemi (MSS), dejeneratif hastalıklar ve retina27,28,29dahil ile ilişkili Microglia hücrelerdir. Böylece, mevcut protokolünde kullanılan iki yöntem birleştirerek, microglia dağıtım ve retina dejenerasyonu yapılabilir, hangi izleme hastalık şiddeti kolaylaştırabilir veya tedavi edici etkinliği içinde vivoyaklaşıyor.

<table>
	<tbody>
		<tr>
			<td>Spectralis Imaging system (HRA+OCT)</td>
			<td>Heidelberg Engineering, Germany</td>
			<td>N/A</td>
			<td>ophthalmic imaging platform system</td>
		</tr>
		<tr>
			<td>Heidelberg Eye Explorer</td>
			<td>Heidelberg Engineering, Germany</td>
			<td>N/A</td>
			<td>Version 1.9.13.0</td>
		</tr>
		<tr>
			<td>78D standard ophthalmic non-contact slit lamp lens</td>
			<td>Volk Optical Inc., Ohio, USA</td>
			<td>V78C</td>
			<td></td>
		</tr>
		<tr>
			<td>Spectralis wide angle 55&#176; lens</td>
			<td>Heidelberg Engineering, Germany</td>
			<td>50897-002</td>
			<td></td>
		</tr>
		<tr>
			<td>ultra widefield 102&#176; lens</td>
			<td>Heidelberg Engineering, Germany</td>
			<td>50117-001</td>
			<td></td>
		</tr>
		<tr>
			<td>medetomidine hydrochloride 1 mg/mL (Domitor)</td>
			<td>Provet AG, Lyssach, Switzerland</td>
			<td>Swissmedic Nr. 50&#39;590 - ATCvet: QN05CM91</td>
			<td>anesthetic/analgesic</td>
		</tr>
		<tr>
			<td>ketamine 50mg/ml (Ketalar)</td>
			<td>Parke-Davis, Zurich, Switzerland</td>
			<td>72276388</td>
			<td>anesthetic</td>
		</tr>
		<tr>
			<td>tropicamide 0.5% + phenylephrine HCl 2.5% (Augentropfen mix)</td>
			<td>ISPI, Bern, Switzerland</td>
			<td>N/A</td>
			<td>pupil dilation</td>
		</tr>
		<tr>
			<td>Omnican Insulin-50 0.5 ml G30 0.3 x 12mm</td>
			<td>B. Braun Mesungen AG, Carl-Braun-Stra&#223;e, Germany</td>
			<td>9151125</td>
			<td></td>
		</tr>
		<tr>
			<td>hydroxypropylmethylcellulose (Methocel 2%)</td>
			<td>OmniVision, Neuhausen, Switzerland</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>+4 dpt rigid gas permeable contact lens</td>
			<td>Quantum I, Bausch + Lomb Inc., Rochester, NY</td>
			<td>N/A</td>
			<td>Base Curve: 7.20 to 8.40 mm 
			Diameter: 9.00 / 9.60 / 10.20 mm 
			Power: -25.00 to +25.00 Diopters</td>
		</tr>
		<tr>
			<td>balanced salt solution (BSS)</td>
			<td>Inselspital, Bern, Switzerland</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>silicon forceps</td>
			<td>N/A</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>atipamezole 5 mg/mL (Antisedan)</td>
			<td>Provet AG, Lyssach, Switzerland</td>
			<td>N/A</td>
			<td>&#945;2 adrenergic receptor antagonist</td>
		</tr>
		<tr>
			<td>GraphPad Prism 7</td>
			<td>GraphPad Software, Inc, San Diego, CA, USA</td>
			<td>N/A</td>
			<td>statistical analysis software</td>
		</tr>
	</tbody>
</table>

in vivo imaging of cx3cr1gfp/gfp reporter mice with spectral-domain optical coherence tomography and scanning laser ophthalmoscopy

Spektral etki alanı optik Koherens tomografi (SD-Ekim) ve tarama lazer ophthalmoscopy (SLO), deneysel Oftalmoloji yaygın olarak kullanılır. Mevcut protokolünde ifade fareler yeşil flüoresan protein (gfp) Cx3cr1 organizatörü altında (BALB/c-Cx3cr1gfp/gfp) microglia hücreleri vivo retina görüntü için kullanıldı. Microglia retina ikamet makrofajlar vardır ve birkaç retina hastalıkları1,2,3,4,5,6' karıştığı olmuştur. Bu iletişim kuralı retina B-bir oftalmik görüntüleme platformu sistemi kullanarak inceden inceye gözden geçirmek, SD-Ekim ve microglia hücre dağıtım SLO vivo içindeile Cx3cr1gfp/gfp farelerde görüntüleme ile üretimi için detaylı bir yaklaşım sağlar. İletişim kuralı birkaç muhabir fare satırlarında kullanılabilir. Ancak, burada sunulan protokolü için bazı sınırlamalar vardır. İlk olarak, SLO hem SD-Ekim, yüksek çözünürlüklü modunda kullanıldığında yüksek Aksiyel çözünürlük ama yanal çözünürlük ile veri toplama düşüktür (3,5 µm ve 6 µm, sırasıyla). Ayrıca, SLO odak ve doygunluk düzeyinde son derece parametre seçimi ve doğru hizalama göz bağlıdır. Ayrıca, insan hasta farelerde yüksek toplam optik güç fare göz nedeniyle zorlu için tasarlanan aygıt kullanarak insan gözünün karşılaştırıldığında; Bu büyütme yanlışlıklar7, lateral hangi da büyütme diğerleri arasında fare objektif tarafından bağlı yol açabilir. Ancak, eksenel tarama pozisyon karşın yanal büyütme bağlıdır, eksenel SD-Ekim ölçümleri doğru8.

Burada sunulan iletişim kuralını kullanarak, SD-Ekim tarar ve SLO görüntüleri Cx3cr1gfp/gfp fareler aynı görüntüleme oturumunda elde edilmiştir. Şekil 3 temsilcisi SD-Ekim 30 ° veya bir 55 ° lens (şekil 3A) ve temsilcisi SLO görüntüleri ile elde edilen tek inceden inceye gözden geçirmek nerede gfp olumlu microglia hücreleri görselleştirildiği 55 ° veya bir 102 ° lens ile elde içerir...

Watch this Scientific Journal Video about In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy at JoVE.com

In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy

Bu protokolü nasıl yüksek çözünürlüklü spektral etki alanı optik Koherens tomografi gibi görüntüleme teknikleri açıklar ve lazer ophthalmoscopy tarama bir oftalmik görüntüleme platformu üzerinde bilgi edinme sistemiyle küçük Rodents yararlı olabilir dağıtım, hücre sırasıyla retina kalınlığı ve mikroglial.

Vivo Tayf alanlı optik Koherens tomografi ve lazer Ophthalmoscopy tarama Cx3cr1gfp/gfp muhabir farelerin görüntüleme

Spectral domain optical coherence tomography (SD-OCT) and scanning laser ophthalmoscopy (SLO) are extensively used in experimental ophthalmology. In the ...

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Medicine

In Vivo Imaging of Cx3cr1gfp/gfp Reporter Mice with Spectral-domain Optical Coherence Tomography and Scanning Laser Ophthalmoscopy

Oral Biofilm Analysis of Palatal Expanders by Fluorescence In-Situ Hybridization and Confocal Laser Scanning Microscopy

Floresan In-Situ Hibridizasyon ve Konfokal Lazer Tarama Mikroskopi Palatal Genişleticilerden Oral Biyofilm Analizi

Confocal laser scanning microscopy (CLSM) of natural heterogeneous biofilm is today facilitated by a comprehensive range of staining techniques, one of ...

Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas

Intracranial Implantation with Subsequent 3D In Vivo Bioluminescent Imaging of Murine Gliomas

Müteakip 3D ile İntrakranial İmplantasyonu In Vivo Murin Gliomlar Bioluminescent Görüntüleme

The mouse glioma 261 (GL261) is recognized as an in vivo model system that recapitulates many of the features of human glioblastoma multiforme (GBM). The ...

Doppler Optical Coherence Tomography of Retinal Circulation

Retina Sirkülasyon Doppler Optik Koherens Tomografi

Noncontact retinal blood flow measurements are performed with a Fourier domain optical coherence tomography (OCT) system using a circumpapillary double ...

Segmentation and Measurement of Fat Volumes in Murine Obesity Models Using X-ray Computed Tomography

Segmentasyon ve Mürin Obezite Modelleri Yağ Hacimleri ölçümü X-ray Bilgisayarlı Tomografi Kullanımı

Obesity is associated with increased morbidity and mortality as well as reduced metrics in quality of life.1 Both environmental and genetic factors are ...

Generation and 3-Dimensional Quantitation of Arterial Lesions in Mice Using Optical Projection Tomography

Üretim ve Farelerde arteriyel Lezyonlarının 3-Boyutlu kantitasyonu Optik Projeksiyon Tomografi ile

The generation and analysis of vascular lesions in appropriate animal models is a cornerstone of research into cardiovascular disease, generating ...

In Vivo Optical Imaging of Brain Tumors and Arthritis Using Fluorescent SapC-DOPS Nanovesicles

In Vivo Optical Imaging of Brain Tumors and Arthritis Using Fluorescent SapC-DOPS Nanovesicles

Floresan SapC-DOPS Nanovesicles kullanma Beyin Tümörleri ve Artrit Vivo Optik Görüntüleme

We describe a multi-angle rotational optical imaging (MAROI) system for in vivo monitoring of physiopathological processes labeled with a fluorescent ...

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Non-invasive Imaging and Analysis of Cerebral Ischemia in Living Rats Using Positron Emission Tomography with 18F-FDG

Pozitron Emisyon Tomografi ile kullanma Non-invaziv Görüntüleme ve Serebral iskemi Analizi Yaşam Sıçanlarda<sup&gt; 18</sup&gt; F-FDG

Stroke is the third leading cause of death among Americans 65 years of age or older1. The quality of life for patients who suffer from a stroke fails to ...

Combined In vivo Optical and &#181;CT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Combined In vivo Optical and Ãƒâ€šÃ‚ÂµCT Imaging to Monitor Infection, Inflammation, and Bone Anatomy in an Orthopaedic Implant Infection in Mice

Kombine<em&gt; In vivo</em&gt; Optik ve uCT Görüntüleme Farelerde bir Ortopedik İmplant Enfeksiyon Enfeksiyon, Enflamasyon ve Kemik Anatomisi Monitör

Multimodality imaging has emerged as a common technological approach used in both preclinical and clinical research. Advanced techniques that combine in ...

Assessment of Viability of Human Fat Injection into Nude Mice with Micro-Computed Tomography

Mikro-Bilgisayarlı Tomografi ile Çıplak Fareler içine İnsan Yağ Enjeksiyonu Canlılığına Değerlendirilmesi

Lipotransfer is a vital tool in the surgeon&#8217;s armamentarium for the treatment of soft tissue deficits of throughout the body. Fat is the ideal soft ...

Validation of Nanobody and Antibody Based In Vivo Tumor Xenograft NIRF-imaging Experiments in Mice Using Ex Vivo Flow Cytometry and Microscopy

Validation of Nanobody and Antibody Based In Vivo Tumor Xenograft NIRF-imaging Experiments in Mice Using Ex Vivo Flow Cytometry and Microscopy

Nanobody ve Antikor Temelli Doğrulama<em&gt; İn Vivo</emFarelerde&gt; Tümör Xenograftlarında NIRF-görüntüleme Deneyler kullanma<em&gt; Ex vivo</em&gt; Sitometrisi ve Mikroskopi

This protocol outlines the steps required to perform ex vivo validation of in vivo near-infrared fluorescence (NIRF) xenograft imaging experiments in mice ...