Wir erkennen Drs. Camillo Ricordi, Antonello Pileggi, R. Damaris Molano, Stephan Speier und Daniel Nyqvist für fruchtbare Diskussionen. Wir danken auch Eleut Hernandez und Diego Espinosa-Heidmann für die technische Unterstützung und Mike Valdes und Margaret Formoso um Hilfe mit Videoaufnahme. Byron Maldonado erfasst, bearbeitet und produziert das fertige Video. Unterstützung der Forschung wurde von der Diabetes Research Institute Foundation (vorausgesetzt <a href="http://www.DiabetesResearch.org" target="_blank">www.DiabetesResearch.org</a> ), der NIH / NIDDK / NIAID (F32DK083226 MHA; NIH RO3DK075487 zum AC; U01DK089538 um PO.B.). Weitere wissenschaftliche Unterstützung PO.B wurde durch Mittel aus dem Karolinska Institutet, dem schwedischen Research Council, dem schwedischen Diabetes Foundation, Familie Erling Persson-Stiftung, der Familie Knut and Alice Wallenberg Foundation, der Skandia Insurance Company Ltd VIBRANT vorgesehen ( FP7-228.933-2), Strategic Research Program in Diabetes am Karolinska Inst.itutet der Novo Nordisk Stiftung und der Berth von Kantzow Stiftung.

Das folgende Verfahren unter dem Stereoskop wird in 2 Schritten durchgeführt wird, beinhaltet der erste Schritt das Laden der Inseln in die Kanüle und der zweite Schritt ist die tatsächliche Transplantation in das ACE. Alle Verfahren an Tieren durchgeführt wurden durch die institutionelle Pflege der Tiere und die Nutzung Ausschuss (IACUC) der University of Miami zugelassen. Ein. Lädt Islets in Kanüle für Transplantation <ol><li> Center die Inseln in Kulturschale durch Drehen der Schüssel in Verengung Kreisen. </li><li> Trennen Sie die Kanüle aus dem &quot;Reservoir&quot; und legen Sie die Kanüle und Verbindungsschlauch auf eine saubere Oberfläche. Der Behälter kann aus einem 300 ul Einweg-Kunststoff-Pipettenspitze werden ohne Filter (Abbildung 1a). </li><li> Bündig Luftblasen aus dem Reservoir zu kontinuierlichen Strom von Inseln sicherzustellen beim Ansaugen in das Reservoir. Spülen des Reservoirs durch Vorantreiben der Freisprech-motorisierten Spritze Treiber mit dem Fußpedal getan(Abbildung 1b, c). Damit wird auch Platz in der Spritze, um eine Aspiration der Inseln in den Vorratsbehälter (mit steriler Lösung wie Kochsalzlösung, PBS oder Kulturmedien vorbelastet) zu ermöglichen. </li><li> Vorsichtig absaugen gewünschten Menge von Inseln in das Reservoir. Inseln werden zu wirbeln neigen, da sie das Reservoir und geben bleiben gemeinsam auf dem Boden. Aspiration wird durch Rückwärtsfahrt den motorisierten Spritze Treiber mit dem Fußpedal getan. </li><li> Schließen Sie die Kanüle in den Behälter über den Verbindungsschlauch. </li><li> Platzieren Sie die Kanülenspitze wieder in der Kulturschale und bündig die Inseln aus dem Reservoir in den Schlauch dann in die Kanüle. Sicherstellen, dass die Inseln zusammen bleiben, wie Sie-back füllen den Schlauch / Kanüle sanft &quot;schnippen&quot; (Tapping) den Schlauch (Abbildung 1d). Stoppen entweder vor oder nach allen Luftblasen vor der Inseln sind die Kanüle gespült. Wenn nicht sicher, stoppen, wie Inseln auf der Rückseite des Kanüle eingeben, wie die restliche LuftBlasen vor der Inseln kann verhindern, Reflux (Rückfluss) von Inseln aus der ACE. Über Nacht zu zerstreuen. </li><li> In diesem Stadium sind Sie bereit, um die Inseln in der ACE (siehe nächste Schritte) zu injizieren. </li></ol> 2. Inseltransplantation in die Vorderkammer des Auges <ol><li> Positionieren Sie den narkotisierten Maus an einem warmen Pad unter Stereoskop. </li><li> Legen Sie die Schnauze der Maus in Narkose &quot;Maske&quot; mit Sauerstoff / Isofluran-Narkose Maschine. Die Maske ist aus einer 1 ml Einweg-Kunststoff-Pipettenspitze (ohne Filter) und mit der Narkose Schlauch durch das schmale Ende (Abbildung 2a, b). </li><li> Vorsichtig einfahren Augenlider des Auges transplantiert mit dem Zeigefinger und Daumen der freien Hand und &quot;Pop&quot; das Auge für eine bessere Belichtung und einfachen Zugang (Abbildung 2c) werden. Dies erfordert einige Übung, um ohne Behinderung Atmung der Maus durch übermäßigen Druck auf die perfekteHalses oder Blockierung des Blutflusses zum Kopf. </li><li> Verwendung eines wegwerfbaren Insulinspritze (29 - 31G) als Skalpell sorgfältig dringen nur die Spitze in der Hornhaut und einen einzigen seitlichen Schnitt. Machen den Einschnitt in der Mitte zwischen dem Scheitelpunkt der Hornhaut und Limbus zum Rückfluss der Inselchen während der Injektion aus der ACE (Abbildung 2 d) zu minimieren. </li><li> Schieben Sie die Kanüle (vorinstalliert mit Inseln) durch den Einschnitt. </li><li> Langsam auszuwerfen Inselchen aus der Kanüle und hinterlegen sie auf der Iris. Um islet Reflux aufgrund übermäßigen Druckaufbau in der ACE zu vermeiden, werfen Sie die Inseln in kurzen Schüben in möglichst wenig Volumen (s) wie möglich in den Quadranten gegenüber dem Einschnitt. Dies kann weiter durch Verdichten Inselchen im Schlauch beim Laden der Kanüle (siehe Schritt 1,6) sichergestellt werden. </li><li> Langsam einfahren Kanüle aus der ACE. Dies ist ein kritischer Schritt, vor allem, wenn ein großes Volumen von Inseln als Insel Rückfluß durch Druck injiziert wurdedass sich im Inneren des ACE unvermeidlich sein. Zu beseitigen / minimieren Insel Reflux, drehen Sie ihn die Kanüle während im Inneren des ACE Überdruck durch den Einschnitt um die Kanüle freizugeben. Auf Anzeichen von Reflux, wie Sie die Kanüle zurückziehen und, falls erforderlich, bis der Druck nachlässt warten, bevor vollständig Zurückziehen der Kanüle aus dem ACE versuchen. </li><li> Spülen Sie den transplantierten Auge mit sterilem PBS oder Kochsalzlösung. </li><li> Injizieren Buprenorphin zur postoperativen Analgesie (0,05 bis 0,1 mg / kg, subkutan) für die ersten 48 Stunden. </li><li> Bewerben Erythromycin Augenheilkunde antibiotische Salbe auf die transplantierten Auge sofort nach der Transplantation. </li><li> Legen Sie das Tier wieder in einer vorgewärmten Käfig aus der Narkose ermöglichen. </li></ol>

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PO.B. ist einer der Gründer des Biotech-Unternehmens Biocrine, die gehen, um die Vorderkammer des Auges als kommerzielles Service-Plattform zu verwenden. AC ist auf dem Patent zum Schutz dieser Technologie.

Murine Pankreasinseln wurden isoliert unter Verwendung Kollagenaseverdau durch Reinigung auf Dichtegradienten gefolgt, wie zuvor beschrieben 33. Isolierten Inseln wurden über Nacht vor der Transplantation gezüchtet. Obwohl dies nicht erforderlich sein kann, wird empfohlen, damit die Inseln aus dem Isolierungsverfahren erholen. Dies ist entscheidend, wenn die Transplantation in diabetischen Empfängern durchgeführt wird, wie es der Transplantation zu überleben / robust Inseln gewährleistet. <p class="...

Advances in Intravitalmikroskopie haben physiologischen Erscheinungen nicht vorhergesagt durch in vitro Studien 1 offenbart. Dies unterstreicht die Herausforderung bei der Umsetzung Erkenntnisse durch konventionelle In-vitro-Methoden erhalten in den lebenden Tier. In den letzten zehn Jahren wurde das Sichtbarmachen von Geweben in lebenden Tieren erheblich durch technologische Fortschritte in bildgebenden Verfahren 2, 3, 4, 5, 6 verbessert. Dies hat einen Bedarf für in vivo Bildgebung Ansätze mit mögliches Einsatzgebiet in experimentellen Tiermodellen zu longitudinalen Visualisierung von Zielgeweben nichtinvasiv ermöglichen vorangetrieben. Bildgebende Verfahren wie Kernspintomographie und Positronen-Emissions-Tomographie oder Biolumineszenz haben nicht-invasive Bildgebung von Organen / Geweben tief innerhalb des Körpers 7-8, 9 aktiviert. Aber diese Techniken nicht erreichen können einzelne Zelle Auflösung aufgrund hoher Hintergrundsignale und niedriger räumlicher Auflösung, trotz der Verwendung of Hochkontrastmaterialien oder gewebespezifische Lumineszenz 4. Dies wurde mit dem Aufkommen der Zwei-Photonen-Fluoreszenz-Konfokalmikroskopie 10 gerichtet. Zwei-Photonen-Mikroskopie aktiviert intravital Bildgebung zur Visualisierung und Quantifizierung von zellulären Ereignissen mit beispielloser Details 11, 12. Dies hat zur Charakterisierung von wichtigen biologischen Prozessen in Gesundheit und Krankheit 13, 14, 15, 16 geführt. Während bahnbrechenden intravital bildgebenden Untersuchungen in erster Linie &quot;nachgeahmt&quot; in-vivo-Bedingungen in herausgeschnittene Gewebe (zB Lymphknoten), haben andere Studien invasive Ansätze zur Bild belichtet Zielgewebe verwendet in situ 17, 18, ​​19, 20, 21. Andere Studien haben auch &quot;Fenster Kammer Modelle&quot; Einschränkungen bei invasiven Konzepten und begrenzte Bildauflösung in vivo 22, 23, 24, 25 zugeordnet zu umgehen verwendet. In dem Fenster Kammer Modell wird eine Kammer mit einem transparenten Fenster chirurgisch in die Haut an verschie implantiertMiete Standorten (dorsal oder Ohr haut, Brustfettpolster, Leber, etc.) auf das Tier (zB Maus, Ratte, Kaninchen). Während dieser Ansatz ermöglicht eindeutig hochauflösenden in vivo Bildgebung erfordert es eine invasive Chirurgie, um die Kammer zu implantieren und ist möglicherweise nicht in der Lage, longitudinalen Bildgebungsstudien über mehrere Wochen oder Monate 22 unterzubringen. Es wurde kürzlich gezeigt, dass die Kombination von hoher Auflösung konfokale Mikroskopie mit einem minimal-invasiven Eingriff, nämlich Transplantation in die vordere Kammer des Auges (ACE) eine &quot;natürliche Körper Fenster&quot; als eine leistungsfähige und vielseitige in vivo Bildgebung Plattform 26, 27. Transplantation in das ACE hat sich in den letzten Jahrzehnten eingesetzt, um biologische Aspekte einer Vielzahl von Geweben 28, 29, 30 zu studieren, und seine jüngsten Kombination mit hochauflösenden bildgebenden aktiviert das Studium der Physiologie der Langerhans-Inseln mit Einzel-Zell-Auflösung nicht invasiv und längs &lt;sup&gt; 26, 27. Dieser Ansatz wurde verwendet, um Autoimmunreaktionen bei der Entwicklung von Typ 1 Diabetes im Tiermodell (unveröffentlichte Daten) zu studieren. Es wurde auch verwendet, um Bauchspeicheldrüsenkrebs Entwicklung zu studieren, sowie in Studien der Nierenfunktion durch Transplantation in die ACE Pankreas Knospen oder einzelne Nierenglomeruli bzw. (unveröffentlichte Daten). Ein kürzlich veröffentlichter Bericht über diesen Ansatz weiter demonstriert seine Anwendung auf Immunantwort nach Inselzellen der Bauchspeicheldrüse Transplantation 31 studieren. Wichtig ist, dass diese Studie zeigte, dass die Transplantation in die vordere Kammer des Auges stellt eine natürliche Körper Fenster ausführen: (1) Längs-, nicht-invasiven Darstellung von transplantierten Geweben in vivo, (2) in vivo cytolabeling um zelluläre Phänotyp und Lebensfähigkeit in beurteilen situ, (3) Echtzeit-Tracking von infiltrierenden Immunzellen in das Zielgewebe, und (4) lokale Intervention durch topische Anwendung oder intraokularen Injektion. Hier D Wiremonstrate wie Transplantation in die vordere Kammer des Auges unter Verwendung Pankreasinseln durchzuführen.

<table border="1"><tbody><tr><td> Namen Reagenz </td><td> Firma </td><td> Katalog-Nummer </td><td> Beschreibung / Kommentare </td></tr><tr><td> IsoTHESIA (Isofluran) </td><td> Buttler Animal Health Versorgung </td><td> 11695-6775-2 </td><td> 99,9% Isofluran / ml </td></tr><tr><td> Ketaset (Ketamin HCL) </td><td> Fort Dodge Animal Health </td><td> 0856-2013-01 </td><td> Alternative injizierbaren Anästhesie </td></tr><tr><td> Beprenex (Buprenorphin HCL) </td><td> Reckitt Benckiser Health Care (UK) Ltd </td><td> 12496-075-7-1 </td><td> 0,3 mg / ml </td></tr><tr><td> Erythromycin Augensalbe USP, 0,5% </td><td> Akron </td><td> 17478-070-35 </td><td> Prophylaktisch an transplantierten Auge angewendet </td></tr><tr><td> 0,9% Natriumchlorid (Saline) </td><td> Hospira Inc. </td><td> 0409-7983-03 </td><td> Für iv Injektion. Steril </td></tr><tr><td> PBS </td><td> Gibco </td><td> 10010-023 </td><td> 1X. Steril </td></tr><tr><td> CMRL Medium 1066 </td><td> Cellgro </td><td> 98-304-CV </td><td> Ergänzt, CIT Modifikation. Bevorzugte Medien für Inselchen </td></tr></tbody></table>

transplantation into the anterior chamber of the eye for longitudinal, non-invasive in vivo imaging with single-cell resolution in real-time

Intravital Imaging hat ein unverzichtbares Werkzeug in der biologischen Forschung entstanden. Bei dem Verfahren wurden viele bildgebende Verfahren entwickelt worden, um verschiedene biologische Prozesse bei Tieren nicht-invasiv zu studieren. Allerdings ist eine große technische Einschränkung in bestehende intravital bildgebenden Verfahren die Unfähigkeit, nicht-invasive, Längs-Bildgebung mit Single-Cell Auflösungsvermögen kombinieren. Wir zeigen hier, wie die Transplantation in der Vorderkammer des Auges, erhebliche Einschränkung bietet eine vielseitige experimentelle Plattform, die nicht-invasive, Längs-Bildgebung mit zellulärer Auflösung in vivo ermöglicht umgeht. Wir zeigen die Transplantation Verfahren in der Maus und bieten repräsentative Ergebnisse mit Hilfe eines Modells mit klinischer Relevanz, nämlich Inselzellen der Bauchspeicheldrüse Transplantation. Zusätzlich zur Aktivierung direkten Visualisierung in einer Vielzahl von Geweben in die vordere Kammer des Auges transplantiert, bietet dieser Ansatz eine Plattform, um Gerölln Medikamenten indem Langzeit-Follow up und Überwachung im Zielgewebe. Wegen ihrer Vielseitigkeit, Gewebe / Zell-Transplantation in die vordere Kammer des Auges nicht nur Vorteile Transplantationstherapien, erstreckt sie auf andere in vivo-Anwendungen der physiologischen und pathophysiologischen Prozessen wie Signaltransduktion und Krebs oder Autoimmunerkrankung Entwicklung zu studieren.

Es gibt ein paar Parameter, die eine &quot;gute&quot; Transplantation zu definieren. Eine gute Transplantation ist eine, die läuft ohne Blutungen bei dem Schnitt wie in dem Video zu sehen. Blutungen verhindert / durch Eindringen nur die Spitze des Skalpells (Nadel) in die ACE (Abb. 3a) minimiert. Dies wird auch dazu beitragen, Kontakt und Punktion der Iris. Es wird auch dafür sorgen, einen kleinen Einschnitt, die sehr gut heilen, ohne dass Trübungen der Hornhaut im Laufe der Zeit (Abbildung 3...

Watch this Scientific Journal Video about Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time at JoVE.com

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

Ein neuer Ansatz, der intraokularen Transplantation und konfokale Mikroskopie ermöglicht Längs-, non-invasive Bildgebung in Echtzeit mit Single-Cell Auflösung innerhalb gepfropft Geweben<em&gt; In vivo</em&gt;. Wir zeigen, wie Langerhans-Inseln in die vordere Kammer der Maus Auge zu transplantieren.

Transplantation in die Vorderkammer des Auges für Längs-, Non-invasive<em&gt; In vivo</em&gt; Imaging mit Single-cell Auflösung in Echtzeit

Intravital imaging has emerged as an indispensable tool in biological research.  In the process, many imaging techniques have been developed to study ...

transplantation-into-anterior-chamber-eye-for-longitudinal-non

Research

JoVE Journal

Medicine

Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

13.3K Aufrufe.  University of Miami Miller School of Medicine. Ein neuer Ansatz, der intraokularen Transplantation und konfokale Mikroskopie ermöglicht Längs-, non-invasive Bildgebung in Echtzeit mit Single-Cell Auflösung innerhalb gepfropft Geweben In vivo. Wir zeigen, wie Langerhans-Inseln in die vordere Kammer der Maus Auge zu transplantieren.

Serie: Transplantation into the Anterior Chamber of the Eye for Longitudinal, Non-invasive In vivo Imaging with Single-cell Resolution in Real-time

High-frequency High-resolution Echocardiography: First Evidence on Non-invasive Repeated Measure of Myocardial Strain, Contractility, and Mitral Regurgitation in the Ischemia-reperfused Murine Heart

Ischemia-reperfusion (IR) was surgically performed in murine hearts which were then subjected to repeated imaging to monitor temporal changes in functional parameters of key clinical significance. Two-dimensional movies were acquired at high frame rate (8 kHz) and were utilized to estimate high-quality myocardial strain. Two-dimensional elastograms (strain images), as well as strain profiles, were visualized. Results were powerful in quantitatively assessing IR-induced changes in cardiac events including left-ventricular (LV) contraction, LV relaxation and isovolumetric phases of both pre-IR and post-IR beating hearts in intact mice. In addition, compromised sector-wise wall motion and anatomical deformation in the infarcted myocardium were visualized.  The elastograms were uniquely able to provide information on the following parameters in addition to standard physiological indices that are known to be affected by myocardial infarction in the mouse: internal diameters of mitral valve orifice and aorta, effective regurgitant orifice, myocardial strain (circumferential as well as radial), turbulence in blood flow pattern as revealed by the color Doppler movies and velocity profiles, asynchrony in LV sector, and changes in the length and direction of vectors demonstrating slower and asymmetrical wall movement. This work emphasizes on the visual demonstration of how such analyses are performed. 

High-frequency High-resolution Echocardiography: First Evidence on Non-invasive Repeated Measure of Myocardial Strain, Contractility, and Mitral Regurgitation in the Ischemia-reperfused Murine Heart 

High frequency Doppler ultrasound is a novel technology for assessing regional myocardial function. This work presents first evidence demonstrating applicability of this versatile imaging platform for the repeated measure of myocardial strain, dp/dt, and mitral regurgitation in the ischemia-reperfused (IR) murine heart.

Hochfrequenz-High-Resolution Echokardiographie: First Evidence on Non-invasive Wiederholte Messung der myokardialen Strain, Kontraktilität und Mitralinsuffizienz in der Ischämie-Reperfusion Murine Herz

Ischemia-reperfusion (IR) was surgically performed in murine hearts which were then subjected to repeated imaging to monitor temporal changes in ...

Forschung

Medizin

Multispectral Real-time Fluorescence Imaging for Intraoperative Detection of the Sentinel Lymph Node in Gynecologic Oncology

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical excision of the tumor with a full lymphadenectomy in the drainage area of the tumor. However, removal of lymph nodes is associated with increased morbidity due to infection, wound breakdown and lymphedema.4,5 As an alternative, the sentinel lymph node procedure (SLN) was developed several decades ago to detect the first draining lymph node from the tumor.6 In case of lymphogenic dissemination, the SLN is the first lymph node that is affected (Figure 1). Hence, if the SLN does not contain metastases, downstream lymph nodes will also be free from tumor metastases and need not to be removed. The SLN procedure is part of the treatment for many tumor types, like breast cancer and melanoma, but also for cancer of the vulva and cervix.7 The current standard methodology for SLN-detection is by peritumoral injection of radiocolloid one day prior to surgery, and a colored dye intraoperatively. Disadvantages of the procedure in cervical and vulvar cancer are multiple injections in the genital area, leading to increased psychological distress for the patient, and the use of radioactive colloid.
Multispectral fluorescence imaging is an emerging imaging modality that can be applied intraoperatively without the need for injection of radiocolloid. For intraoperative fluorescence imaging, two components are needed: a fluorescent agent and a quantitative optical system for intraoperative imaging. As a fluorophore we have used indocyanine green (ICG). ICG has been used for many decades to assess cardiac function, cerebral perfusion and liver perfusion.8 It is an inert drug with a safe pharmaco-biological profile. When excited at around 750 nm, it emits light in the near-infrared spectrum around 800 nm. A custom-made multispectral fluorescence imaging camera system was used.9.
The aim of this video article is to demonstrate the detection of the SLN using intraoperative fluorescence imaging in patients with cervical and vulvar cancer. Fluorescence imaging is used in conjunction with the standard procedure, consisting of radiocolloid and a blue dye. In the future, intraoperative fluorescence imaging might replace the current method and is also easily transferable to other indications like breast cancer and melanoma.

Fluorescence imaging is a promising innovative modality for image-guided surgery in surgical oncology. In this video we describe the technical procedure for detection of the sentinel lymph node using fluorescence imaging as showcased in gynecologic oncologicy. A multispectral fluorescence camera system, together with the fluorescent agent indocyanine green, is applied.

Multispectral Real-time Fluoreszenz-Imaging für die intraoperative Detektion des Sentinel Lymph Node in Gynäkologische Onkologie

The prognosis in virtually all solid tumors depends on the presence or absence of lymph node metastases.1-3 Surgical treatment most often combines radical ...

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

The number of patients with end-stage renal disease, and the number of kidney allograft recipients continuously increases. Episodes of acute cellular allograft rejection (AR) are a negative prognostic factor for long-term allograft survival, and its timely diagnosis is crucial for allograft function 1. At present, AR can only be definitely diagnosed by core-needle biopsy, which, as an invasive method, bares significant risk of graft injury or even loss. Moreover, biopsies are not feasible in patients taking anticoagulant drugs and the limited sampling site of this technique may result in false negative results if the AR is focal or patchy. As a consequence, this gave rise to an ongoing search for new AR detection methods, which often has to be done in animals including the use of various transplantation models.
Since the early 60s rat renal transplantation is a well-established experimental method for the examination and analysis of AR 2. We herein present in addition small animal positron emission tomography (PET) using 18F-fluorodeoxyglucose (FDG) to assess AR in an allogeneic uninephrectomized rat renal transplantation model and propose graft FDG-PET imaging as a new option for a non-invasive, specific and early diagnosis of AR also for the human situation 3. Further, this method can be applied for follow-up to improve monitoring of transplant rejection 4.

Non-invasive Imaging of Acute Allograft Rejection after Rat Renal Transplantation Using 18F-FDG PET

We herein present a rat renal transplantation model to non-invasively assess acute allograft rejection using positron emission tomography with 18F-fluorodeoxyglucose.

Nicht-invasive Bildgebung des akuten Abstoßung nach Nierentransplantation mit Rat<sup&gt; 18</sup&gt; F-FDG PET

The number of patients with end-stage renal  disease, and the number of kidney allograft recipients continuously increases.  Episodes of acute cellular ...

Development of an Audio-based Virtual Gaming Environment to Assist with Navigation Skills in the Blind

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio based cues and set within the context of a video game metaphor, users gather relevant spatial information regarding a building's layout. This allows the user to develop an accurate spatial cognitive map of a large-scale three-dimensional space that can be manipulated for the purposes of a real indoor navigation task. After game play, participants are then assessed on their ability to navigate within the target physical building represented in the game. Preliminary results suggest that early blind users were able to acquire relevant information regarding the spatial layout of a previously unfamiliar building as indexed by their performance on a series of navigation tasks. These tasks included path finding through the virtual and physical building, as well as a series of drop off tasks. We find that the immersive and highly interactive nature of the AbES software appears to greatly engage the blind user to actively explore the virtual environment. Applications of this approach may extend to larger populations of visually impaired individuals.

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind.

Entwicklung eines Audio-based Virtual Gaming Umwelt mit Navigation Skills in the Blind Assist

Audio-based Environment Simulator (AbES) is virtual environment software designed to improve real world navigation skills in the blind. Using only audio ...

Chemotherapy-induced Vascular Toxicity - Real-time In vivo Imaging of Vessel Impairment

Certain classes of chemotherapies may exert acute vascular changes that may progress into long-term conditions that may predispose the patient to an increased risk of vascular morbidity.&#160;Yet, albeit&#160;the mounting clinical evidence, there is a paucity of clear studies of vascular toxicity and therefore the etiology of a heterogeneous group of vascular/cardiovascular disorders remains to be elucidated. Moreover, the mechanism that may underlie vascular toxicity can completely differ from the principles of chemotherapy-induced cardiotoxicity, which is related to direct myocyte injury. We have established a real-time, in vivo molecular imaging platform&#160;to evaluate&#160;the potential&#160;acute&#160;vascular toxicity of anti-cancer therapies.
We have set up a platform of in vivo, high-resolution molecular imaging&#160;in mice,&#160;suitable for visualizing vasculature within confined organs and reference blood vessels within the same individuals whereas each individual serve as its own control. Blood vessel walls were impaired after doxorubicin administration, representing a unique mechanism of vascular toxicity that may be the early event in end-organ injury. Herein, the method of fibered confocal fluorescent microscopy (FCFM) based imaging is described, which provides an innovative mode to understand physiological phenomena at the cellular and sub-cellular levels in animal subjects.

Chemotherapy-induced Vascular Toxicity - Real-time In vivo Imaging of Vessel Impairment

We herein describe the method of fibered confocal fluorescent microscopy (FCFM) based imaging, which provides an innovative mode to understand physiological phenomena at the cellular and sub-cellular levels in animal subjects.

Chemotherapie-induzierte vaskuläre Toxizität - Echtzeit-<em&gt; In-vivo-</em&gt; Imaging von Vessel Impairment

Certain classes of chemotherapies may exert acute vascular changes that may progress into long-term conditions that may predispose the patient to an ...

Non-invasive Assessment of the Efficacy of New Therapeutics for Intestinal Pathologies Using Serial Endoscopic Imaging of Live Mice

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic mechanisms that contribute to the onset and progression of intestinal diseases. The identification of new molecules that promote these pathologies has led to a flurry of activity focused on the development of potential new therapies to inhibit their function. As a result, various pre-clinical mouse models with an intact immune system and stromal microenvironment are now heavily used. Here we describe three experimental protocols to test the efficacy of new therapeutics in pre-clinical models of (1) acute mucosal damage, (2) chronic colitis and/or colitis-associated colon cancer, and (3) sporadic colorectal cancer. We also outline procedures for serial endoscopic examination that can be used to document the therapeutic response of an individual tumor and to monitor the health of individual mice. These protocols provide complementary experimental platforms to test the effectiveness of therapeutic compounds shown to be well tolerated by mice.

We describe methods for longitudinal monitoring of the efficacy of therapeutics for the treatment of colonic pathologies in mice using a rigid endoscope. This protocol can be readily used for the characterization of the therapeutic response of an individual tumor in live mice and also for monitoring potential disease relapse.

Nicht-invasive Beurteilung der Wirksamkeit neuer Therapeutika für Darmpathologien Verwenden der seriellen endoskopische Bildgebung von lebenden Mäusen

Animal models of inflammatory bowel disease (IBD) and colorectal cancer (CRC) have provided significant insight into the cell intrinsic and extrinsic ...

Encapsulation Thermogenic Preadipocytes for Transplantation into Adipose Tissue Depots

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular weight metabolites in tissues of the treated host to achieve long-term survival. The semipermeable membrane allows engrafted encapsulated cells to avoid rejection by the immune system. The encapsulation procedure was designed to enable a controlled release of bioactive compounds, such as insulin, other hormones, and cytokines. Here we describe a method for encapsulation of catabolic cells, which consume lipids for heat production and energy dissipation (thermogenesis) in the intra-abdominal adipose tissue of obese mice. Encapsulation of thermogenic catabolic cells may be potentially applicable to the prevention and treatment of obesity and type 2 diabetes. Another potential application of catabolic cells may include detoxification from alcohols or other toxic metabolites and environmental pollutants.

Here, we present a protocol for encapsulation of catabolic cells, which consume lipids for heat production in intra-abdominal adipose tissue and increase energy dissipation in obese mice.

Encapsulation Thermogenic Präadipozyten für die Transplantation in Fettgewebe Depots

Cell encapsulation was developed to entrap viable cells within semi-permeable membranes. The engrafted encapsulated cells can exchange low molecular ...

Extended Time-lapse Intravital Imaging of Real-time Multicellular Dynamics in the Tumor Microenvironment

In the tumor microenvironment, host stromal cells interact with tumor cells to promote tumor progression, angiogenesis, tumor cell dissemination and metastasis. Multicellular interactions in the tumor microenvironment can lead to transient events including directional tumor cell motility and vascular permeability. Quantification of tumor vascular permeability has frequently used end-point experiments to measure extravasation of vascular dyes. However, due to the transient nature of multicellular interactions and vascular permeability, the kinetics of these dynamic events cannot be discerned. By labeling cells and vasculature with injectable dyes or fluorescent proteins, high-resolution time-lapse intravital microscopy has allowed the direct, real-time visualization of transient events in the tumor microenvironment. Here we describe a method for using multiphoton microscopy to perform extended intravital imaging in live mice to directly visualize multicellular dynamics in the tumor microenvironment. This method details cellular labeling strategies, the surgical preparation of a mammary skin flap, the administration of injectable dyes or proteins by tail vein catheter and the acquisition of time-lapse images. The time-lapse sequences obtained from this method facilitate the visualization and quantitation of the kinetics of cellular events of motility and vascular permeability in the tumor microenvironment.

This protocol describes the use of multiphoton microscopy to perform extended time-lapse imaging of multicellular interactions in real time, in vivo at single cell resolution.

Erweiterte Zeitraffer-Intravital Imaging von Echtzeit-Vielzellige Dynamics im Tumormikromilieu

In the tumor microenvironment, host stromal cells interact with tumor cells to promote tumor progression, angiogenesis, tumor cell dissemination and ...

Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

Remodeling of the brain vasculature is a common trait of brain pathologies. In vivo imaging techniques are fundamental to detect cerebrovascular plasticity or damage occurring overtime and in relation to neuronal activity or blood flow. In vivo two-photon microscopy allows the study of the structural and functional plasticity of large cellular units in the living brain. In particular, the thinned-skull window preparation allows the visualization of cortical regions of interest (ROI) without inducing significant brain inflammation. Repetitive imaging sessions of cortical ROI are feasible, providing the characterization of disease hallmarks over time during the progression of numerous CNS diseases. This technique accessing the pial structures within 250 &#956;m of the brain relies on the detection of fluorescent probes encoded by genetic cellular markers and/or vital dyes. The latter (e.g., fluorescent dextrans) are used to map the luminal compartment of cerebrovascular structures. Germane to the protocol described herein is the use of an in vivo marker of amyloid deposits, Methoxy-O4, to assess Alzheimer&#39;s disease (AD) progression. We also describe the post-acquisition image processing used to track vascular changes and amyloid depositions. While focusing presently on a model of AD, the described protocol is relevant to other CNS disorders where pathological cerebrovascular changes occur.

Longitudinal In Vivo Imaging of the Cerebrovasculature: Relevance to CNS Diseases

This manuscript describes a procedure to track the remodeling of the cerebrovasculature during amyloid plaque accumulation in vivo using longitudinal two-photon microscopy. A thinned-skull preparation enables the visualization of fluorescent dyes to assess the progression of cerebrovascular damage in a mouse model of Alzheimer&#39;s disease.

in Längsrichtung<em&gt; In Vivo</em&gt; Imaging des Cerebrovasculature: Bedeutung für ZNS-Erkrankungen

Remodeling of the brain vasculature is a common trait of brain pathologies. In vivo imaging techniques are fundamental to detect cerebrovascular ...

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. To better understand the mechanism behind these surgical procedures, a rat partial liver transplantation (PLTx) model was established for relevant surgical studies. Because of the complexity of the rat PLTx model, a protocol with detailed descriptions is required. An article published previously reported a protocol in which ex vivo hepatectomy was used to achieve 50% rat PLTx. In contrast to this protocol, we introduced a re-arterialized PLTx with an in vivo 70% hepatectomy. An updated vessel-oriented hepatectomy was incorporated into the rat PLTx to refine the microsurgical procedure. The portal veins and hepatic arteries of the left lateral lobe and the median lobe were individually dissected and ligated before removal of the liver parenchyma, thereby decreasing the probability of bleeding in the remnant liver stump. Furthermore, an end-to-side vessel anastomosis between the common hepatic artery and the enlarged proper hepatic artery was introduced to re-arterialize the hepatic artery. By using this end-to-side vessel anastomosis technique, the diameter of the anastomosis was enlarged, thereby decreasing the difficulty of hand suture and maintaining a high rate of anastomotic patency. Moreover, the cuff anastomosis of the infrahepatic vena cava was slightly modified. A section of circumferential liver parenchyma around the vena cava of a recipient was preserved during cuff anastomosis to maintain the three-dimensional shape of the vascular lumen. This section of liver parenchyma was removed after completing the anastomosis. With this modification, the step involving placement of stay sutures was omitted, thereby further shortening the cuff anastomosis time. By using this protocol of rat PLTx, a low liver enzyme level, an intact liver lobular architecture and a high survival rate were achieved after microsurgery.

Re-Arterialized Rat Partial Liver Transplantation with an in vivo Vessel-Oriented 70% Hepatectomy

Here, a protocol involving re-arterialized rat partial liver transplantation is presented. Specifically, 70% liver was resected in vivo by using an updated technique of vessel-oriented hepatectomy. The hepatic artery was reconstructed in an end-to-side manner. The cuff technique was modified to shorten the anastomosis time of the infrahepatic vena cava.

Re Arterialized Ratte teilweise Leber Transplantation mit einem in Vivo Schiff-orientierte 70 % Hepatektomie

Split liver transplantation and living liver donor liver transplantation were developed in the clinic to utilize liver organs in a more efficient manner. ...