Diese Arbeit wurde von der National Science Foundation Grant No 0651981 und die Alexander und Margaret Stewart Vertrauen obwohl die Cancer Center der Johns Hopkins University gefördert. Die Autoren bedanken sich bei den Beiträgen von Dr. Rhoda Alani dem IRB und der Patient Studie sowie die Hilfe und Unterstützung von Dr. Sewon Kang und seine Abteilung während der Patient Studie zu bestätigen.

1. Setup- <ol><li> Eine temperaturgesteuerte Prüfung Zimmer mit einer Infrarot-Kamera und einem PC für die Infrarot-Bilderfassung und-speicherung sowie eine Messkarte an einen Computer angeschlossen ausgestattet sind in Abb. 1a dargestellt. </li><li> Die Raumtemperatur und die Haut Oberflächentemperatur durch Thermoelemente an ein Datenerfassungssystem Karte während der Patient zu studieren und Messdaten überwacht werden auf dem Computer gespeichert. </li></ol> 2. Image Acquisition <ol><li> Da die Läsion nicht im Wärmebild ohne die Kühlwirkung festgestellt werden, ist ein Quadrat Klebstoff Marker zur Lokalisierung der pigmentierten Läsion des Interesses und seiner Umgebung (Abb. 1b). </li><li> Wir erwerben ein helles Licht Bild der pigmentierten Läsionen und der Klebstoff-Fenster mit einer Digitalkamera (Canon PowerShot G11) (Abb. 1b). </li><li> Ein Dermatoskop verbunden mit einer digitalen Kamera (DermLite Foto System) wird verwendet, um das polarisierte Licht Bilderfassung. </li><li> Wir erwerben ein steady state Infrarot-Bild mit einem Merlin midwave (3-5 um) Infrarot-Kamera in Abb. 1a, c gezeigt </li><li> Wir wenden einen Strom kalter Luft in den Bereich der Haut des Patienten mit der Läsion sowie eine 50 mm Durchmesser Region für die Dauer von einer Minute. </li><li> Nach einer Minute, entfernen wir diese Abkühlung Stress lassen die Haut bei Raumtemperatur innerhalb von 3-4 Minuten (thermische Verwertung Phase) (Abb. 1c-d) wieder warm. </li><li> Während der thermischen Verwertung Phase sind Infrarotaufnahmen der pigmentierten Läsion alle 2 Sekunden (Abb. 1c-d) erfasst. </li><li> Alle IR-Aufnahmen (zusätzlich zu dem weißen Licht und polarisiertes Licht Bilder) während der Studie genommen werden gespeichert und abgelegt unter Verwendung der Labview-Software. </li></ol> 3. Image Processing <ol><li> Die IR-Bilder werden analysiert mit Hilfe eines speziellen Matlab-Code, um genaue transiente Temperaturverteilung auf der Hautoberfläche zu erhalten. Zu diesem Zweck stellen wir mehrere Kalibrierung Schritte und ein multimodales Bild-Analyse-System. </li><li> Wir beginnen mit der Anwendung ein Wahrzeichen Erkennungsalgorithmus an das helle Licht die Grafik für eine Lokalisierung von den Ecken der Klebstoff-Marker. Anschließend identifizierten wir die entsprechenden Punkte in den Referenz-IR-Bild. </li><li> Um für unfreiwillige Körper / Bewegung des Körpers des Patienten auszugleichen, verwenden wir diese Punkte als Landmarken in eine quadratische Bewegung Modell für die Ausrichtung der IR-Bildsequenz während der Erholungsphase. </li><li> Wir verwenden die zufällige Wanderer, ein interaktives Bild Segmentierungsalgorithmus, wo der Benutzer räumlich Leitfaden kann die Segmentierung, indem Samen Punkte, um eine Maske Bild Abgrenzung der Läsion zu schaffen. </li><li> Sobald wir die Form der Läsion zu bestimmen, identifizieren wir die entsprechende Region in jeder der eingetragenen IR-Bilder. </li><li> Wir wählen zufällige Punkte im Inneren der Läsion und weg von der Läsion, die die Läsion und das gesunde Gewebe bzw.. </li><li> Wir vergleichen die instationären thermischen Verhaltens von gesunder Haut und die Reaktion der Läsion. </li><li> Wir bereiten eine Tabelle mit allen Daten: digital, Dermatoskopie, farbkodierte IR-Bilder der Läsion und Umgebung bei Umgebungsbedingungen und 2 Sekunden nach dem Abkühlen Anregung aufgenommen, und der instationären thermischen Verhaltens der Läsion und das gesunde Gewebe. </li></ol> 4. Repräsentative Ergebnisse: <img src="/files/ftp_upload/2679/2679fig1.jpg" alt="Abbildung 1" /> Abbildung 1. a) Die Infrarot-Imaging-System HRIS in der klinischen Studie Raum, b) Foto des größeren Körperoberfläche mit einem Cluster von pigmentierten Läsionen und die Vorlage Rahmen für die Bildgebung, c angewandt) Hinweis Infrarot-Bild der Region bei Raumtemperatur, d) die gleiche Fläche nach dem Abkühlen und e) vergrößerten Ausschnitt des Melanoms Läsion und Umgebung <img src="/files/ftp_upload/2679/2679fig2.jpg" alt="Abbildung 2" /> Abbildung 2. Exam Zimmer mit unserer Wärmebildkamera-System. <img src="/files/ftp_upload/2679/2679fig3.jpg" alt="Abbildung 3" /> Abbildung 3. Cooling der Läsion und die umliegende Hautgewebe durch Einblasen einen Strom von kalter Luft aus einem Vortex-Röhre.

<ol>
	<li>Elder, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Tumor+progression,+early+diagnosis+and+prognosis+of+melanoma.">Tumor progression, early diagnosis and prognosis of melanoma.</a> Acta Oncol. 38, 535-547 (1999).</li><li>Wartman, D., Weinstock, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Are+we+overemphasizing+sun+avoidance+in+protection+from+melanoma.">Are we overemphasizing sun avoidance in protection from melanoma.</a> Cancer Epidemiol Biomarkers Prev. 17, 469-470 (2008).</li><li>Pirtini Cetingul, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Using high resolution infrared imaging to detect melanoma and dysplastic nevi [dissertation]. , (2010).</li><li>Jones, B. F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+reappraisal+of+the+use+of+infrared+thermal+image+analysis+in+medicine.">A reappraisal of the use of infrared thermal image analysis in medicine.</a> IEEE Trans. Med. Imaging. 17, 1019-1027 (1998).</li><li>Anbar, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clinical+thermal+imaging+today-shifting+from+phenomenological+thermography+to+pathophysiologically+based+thermal+imaging.">Clinical thermal imaging today-shifting from phenomenological thermography to pathophysiologically based thermal imaging.</a> IEEE Eng. Med. Biol. Mag. 17, 25-33 (1998).</li><li>Anbar, M., Gratt, B. M., Hong, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Thermology+and+facial+telethermography.+Part+I:+history+and+technical+review.">Thermology and facial telethermography. Part I: history and technical review.</a> Dentomaxillofacial Radiology. 27, 61-67 (1998).</li><li>Jones, B. F., Plassmann, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Digital+infrared+thermal+imaging+of+human+skin.">Digital infrared thermal imaging of human skin.</a> IEEE Eng. Med. Bio. 21, 41-48 (2002).</li><li>Qi, H., Diakides, N. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Infrared imaging in Medicine. , (2007).</li><li>Pirtini Cetingul, M., Herman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Identification+of+skin+lesions+from+the+transient+thermal+response+using+infrared+imaging+technique.">Identification of skin lesions from the transient thermal response using infrared imaging technique.</a> IEEE 5th Int. Symp. on Biomedical Imaging: From Nano to Macro 1-4. , 1219-1222 (2008).</li><li>Cetingul, P. i. r. t. i. n. i., M, ., Herman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantification+of+the+thermal+signature+of+a+melanoma+lesion.">Quantification of the thermal signature of a melanoma lesion.</a> Int. Journal of Thermal Science. 50, 421-431 (2011).</li><li>Pirtini Cetingul, M., Herman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+heat+transfer+model+of+skin+tissue+for+the+detection+of+lesions:+sensitivity+analysis.">A heat transfer model of skin tissue for the detection of lesions: sensitivity analysis.</a> Physics in Medicine and Biology. 55, 5933-5951 (2010).</li><li>Pirtini Cetingul, M., Herman, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Quantitative+evaluation+of+skin+lesions+using+transient+thermal+imaging.">Quantitative evaluation of skin lesions using transient thermal imaging.</a> Proc. Int. Heat Transfer Conf. , (2010).</li></ol>

Die Ergebnisse deuten darauf hin, dass durch die Anwendung der Kühlung betonen wir die Temperaturunterschiede zwischen der Verletzung und des umliegenden gesunden Gewebes verbessert. Auch, weil der kleine Bewegungen des Patienten während der Wärmebildkamera, mussten wir motion tracking richtig Overlay-Bilder zu Temperaturdifferenzen zwischen Referenz-Zustand und die Temperaturverteilung während der thermischen Verwertung Maßnahme. Ohne die Motion-Tracking würden wir nicht in der Lage zu erkennen und messen die Temperaturdifferenz zwischen maligne Läsion und das gesunde Gewebe zu haben. Diese Ergebnisse und die Notwendigkeit einer genauen Motion-Tracking erklären die Schwierigkeiten Ermittler konfrontiert in der Vergangenheit bei dem Versuch, Melanom diagnostiziert mittels IR-Imaging auf steady state Informationen allein und belegen eindeutig die Vorteile der dynamischen thermischen Bildgebung. Es wird darauf hingewiesen, dass die räumliche Auflösung der IR-Kamera (Anzahl der Pixel in der IR Focal Plane Array) kritisch, wenn anspruchsvolle kleine Läsionen ist. Sowohl die räumliche Auflösung und die Temperaturempfindlichkeit der frühen Infrarot-Kameras war begrenzt, die auch Konten für die Schwierigkeiten bei der Erkennung frühzeitig Melanom in der Vergangenheit. Die wichtigsten Unterschiede zwischen unserem Ansatz und vor Wärmebildkameras versucht - das waren mäßig erfolgreich - sind die Sequenzen von Kalibrier-und Bildverarbeitungs-Schritte, die uns genau zu messen Temperaturunterschiede in diesem System zusätzlich zu den dynamischen bildgebenden Verfahren, die auf eine aktive Kühlung angewiesen zu ermöglichen.

<table border="1"><tbody><tr><td> Bezeichnung des Geräts </td><td> Firma </td><td> Katalog-Nummer </td></tr><tr><td> Merlin MWIR Kamera </td><td> FLIR </td><td> - </td></tr><tr><td> Canon PowerShot G11 </td><td> Kanon </td><td> - </td></tr><tr><td> DermLite Foto-System </td><td> DermLite </td><td> - </td></tr><tr><td> Vortex Tube </td><td> EXAIR </td><td> - </td></tr><tr><td> Druckluftbehälter </td><td> Airgas </td><td> - </td></tr></tbody></table>

quantitative visualization and detection of skin cancer using dynamic thermal imaging

Im Jahr 2010 etwa 68.720 Melanome werden allein in den USA diagnostiziert werden, mit rund 8.650 Todesfolge 1. Bis heute ist die einzig wirksame Behandlung des Melanoms chirurgische Exzision, daher der Schlüssel zum Überleben verlängert Früherkennung ist 2,3. Angesichts der großen Zahl von Patienten erkranken jedes Jahr und die Einschränkungen beim Zugriff auf spezialisierte Versorgung schnell, ist die Entwicklung der objektiven In-vivo-Diagnostik-Instrumente für die Diagnose Hilfe von wesentlicher Bedeutung. Neue Techniken, um Hautkrebs, vor allem nicht-invasive Diagnose-Tools, erkennen werden in zahlreichen Labors untersucht. Zusammen mit den chirurgischen Methoden, Techniken wie digitale Fotografie, Dermatoskopie, multispektrale bildgebende Systeme (MelaFind), Laser-basierten Systemen (konfokale Laser-Scanning-Mikroskopie, Laser-Doppler-Perfusion Imaging, Optische Kohärenztomographie), Ultraschall, Kernspintomographie, werden derzeit getestet . Jede Technik bietet einzigartige Vorteile und Nachteile, von denen viele einen Kompromiss zwischen Effizienz und Genauigkeit im Vergleich zu Benutzerfreundlichkeit und Kosten Überlegungen zu stellen. Details über diese Techniken und Vergleiche sind in der Literatur 4. Infrarot (IR)-Bildgebung wurde gezeigt, dass eine nützliche Methode, um die Anzeichen bestimmter Krankheiten durch die Messung der lokalen Hauttemperatur zu diagnostizieren. Es gibt eine große Anzahl von Anzeichen dafür, dass Krankheit oder Abweichung von der normalen Funktionsweise von Änderungen der Temperatur des Körpers begleitet werden, was wiederum Einfluss auf die Temperatur der Haut 5,6. Genaue Daten über die Temperatur des menschlichen Körpers und der Haut kann eine Fülle von Informationen über die Prozesse, die für die Wärmeerzeugung und der Thermoregulation, insbesondere die Abweichung vom normalen Bedingungen, die oft durch Krankheit verursacht. Allerdings IR-Imaging ist nicht weit verbreitet in der Medizin aufgrund der vorzeitigen Nutzung der Technologie 7,8 vor einigen Jahrzehnten erkannt, wenn die Temperatur Messgenauigkeit und die räumliche Auflösung unzureichend waren und anspruchsvolle Bildverarbeitungs-Tools verfügbar waren. Diese Situation änderte sich in den späten 1990er Jahren der 2000er Jahre. Advances in IR Messtechnik, Implementierung von digitalen Bildverarbeitungs-Algorithmen und dynamischen IR-Imaging, das die Wissenschaftler nicht nur die räumliche Analyse ermöglicht, sondern auch die zeitliche thermische Verhalten der Haut 9, erlaubt Durchbrüche in das Feld ein. In unserer Forschung untersuchen wir die Machbarkeit der IR-Imaging, mit theoretischen und experimentellen Untersuchungen kombiniert werden, wie eine kostengünstige, nicht-invasiven, in vivo optische Messtechnik für die Detektion von Tumoren, wobei der Schwerpunkt auf der Vorsorgeuntersuchungen und Früherkennung von Melanomen 10-13 . In dieser Studie zeigen, dass wir Daten in einer Patienten-Studie, in der Patienten, die eine pigmentierte Läsionen besitzen mit einer klinischen Indikation für eine Biopsie für die Bildgebung ausgewählte erhalten. Wir verglichen die Differenz in der thermischen Reaktionen zwischen gesundem und bösartigem Gewebe und verglichen unsere Daten mit Biopsie-Ergebnisse. Wir schlossen daraus, dass die erhöhte Stoffwechselaktivität der Melanom-Läsion durch dynamische Infrarot-Bildgebung erfasst werden kann.

Watch this Scientific Journal Video about Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging at JoVE.com

Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging

Wir haben gezeigt, dass maligne pigmentierte Läsionen mit erhöhter Stoffwechselaktivität erzeugen quantifizierbare Mengen an Wärme und die Messung der transienten thermischen Reaktion der Haut zu einer Abkühlung Anregung ermöglicht quantitative Identifizierung von Melanomen und anderen Hauttumoren (vs. nicht-proliferative Nävi) in einem frühen Stadium der Erkrankung.

Quantitative Visualisierung und Erkennung von Hautkrebs mit Dynamic Thermal Imaging

In 2010 approximately 68,720 melanomas will be diagnosed in the US alone, with around 8,650 resulting in death 1. To date, the only effective treatment ...

quantitative-visualization-detection-skin-cancer-using-dynamic

Nanyang Technological University

Research

JoVE Journal

Medicine

16.7K Aufrufe.  The Johns Hopkins University. Wir haben gezeigt, dass maligne pigmentierte Läsionen mit erhöhter Stoffwechselaktivität erzeugen quantifizierbare Mengen an Wärme und die Messung der transienten thermischen Reaktion der Haut zu einer Abkühlung Anregung ermöglicht quantitative Identifizierung von Melanomen und anderen Hauttumoren (vs. nicht-proliferative Nävi) in einem frühen Stadium der Erkrankung.

Serie: Quantitative Visualization and Detection of Skin Cancer Using Dynamic Thermal Imaging

Dual-mode Imaging of Cutaneous Tissue Oxygenation and Vascular Function

Accurate assessment of cutaneous tissue oxygenation and vascular function is important for appropriate detection, staging, and treatment of many health disorders such as chronic wounds. We report the development of a dual-mode imaging system for non-invasive and non-contact imaging of cutaneous tissue oxygenation and vascular function. The imaging system integrated an infrared camera, a CCD camera, a liquid crystal tunable filter and a high intensity fiber light source. A Labview interface was programmed for equipment control, synchronization, image acquisition, processing, and visualization. Multispectral images captured by the CCD camera were used to reconstruct the tissue oxygenation map. Dynamic thermographic images captured by the infrared camera were used to reconstruct the vascular function map. Cutaneous tissue oxygenation and vascular function images were co-registered through fiduciary markers. The performance characteristics of the dual-mode image system were tested in humans.

A dual-mode imaging system was developed for non-contact assessment of cutaneous tissue oxygenation and vascular function.

Dual-Mode-Imaging von Hautgewebe Oxygenation und Gefäßfunktion

Accurate assessment of cutaneous tissue oxygenation and vascular function is important for appropriate detection, staging, and treatment of many health ...

Forschung

Medizin

The Three-Dimensional Human Skin Reconstruct Model: a Tool to Study Normal  Skin and Melanoma Progression

Most in vitro studies in experimental skin biology have been done in 2-dimensional (2D) monocultures, while accumulating evidence suggests that cells behave differently when they are grown within a 3D extra-cellular matrix and also interact with other cells (1-5). Mouse models have been broadly utilized to study tissue morphogenesis in vivo. However mouse and human skin have significant differences in cellular architecture and physiology, which makes it difficult to extrapolate mouse studies to humans. Since melanocytes in mouse skin are mostly localized in hair follicles, they have distinct biological properties from those of humans, which locate primarily at the basal layer of the epidermis. The recent development of 3D human skin reconstruct models has enabled the field to investigate cell-matrix and cell-cell interactions between different cell types. The reconstructs consist of a "dermis" with fibroblasts embedded in a collagen I matrix, an "epidermis", which is comprised of stratified, differentiated keratinocytes and a functional basement membrane, which separates epidermis from dermis. Collagen provides scaffolding, nutrient delivery, and potential for cell-to-cell interaction. The 3D skin models incorporating melanocytic cells recapitulate natural features of melanocyte homeostasis and melanoma progression in human skin. As in vivo, melanocytes in reconstructed skin are localized at the basement membrane interspersed with basal layer keratinocytes. Melanoma cells exhibit the same characteristics reflecting the original tumor stage (RGP, VGP and metastatic melanoma cells) in vivo. Recently, dermal stem cells have been identified in the human dermis (6). These multi-potent stem cells can migrate to the epidermis and differentiate to melanocytes.

In this report, we describe the three-dimensional skin reconstruct model which mimics human skin in architecture and composition. Melanocyte physiology, melanoma progression and the fate of dermal stem cells have been investigated using the skin reconstruct model. The model is also useful as a preclinical tool for drug assessment.

Die dreidimensionalen humanen Haut Rekonstruieren Modell: ein Tool für normale Haut und Melanom Progression Study

Most in vitro studies in experimental skin biology have been done in 2-dimensional (2D) monocultures, while accumulating evidence suggests that cells ...

Biotechnik

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 management strategies. Orthotopic mouse models of oral cancer have been developed to facilitate the study of factors that impact invasion and serve as model system for evaluating anti-tumor therapeutics. In these systems, visualization of disseminated tumor cells within oral cavity tissues has typically been conducted by either conventional histology or with in vivo bioluminescent methods. A primary drawback of these techniques is the inherent inability to accurately visualize and quantify early tumor cell invasion arising from the primary site in three dimensions. Here we describe a protocol that combines an established model for squamous cell carcinoma of the tongue (SCOT) with two-photon imaging to allow multi-vectorial visualization of lingual tumor spread. The OSC-19 head and neck tumor cell line was stably engineered to express the F-actin binding peptide LifeAct fused to the mCherry fluorescent protein (LifeAct-mCherry). Fox1nu/nu mice injected with these cells reliably form tumors that allow the tongue to be visualized by ex-vivo application of two-photon microscopy. This technique allows for the orthotopic visualization of the tumor mass and locally invading cells in excised tongues without disruption of the regional tumor microenvironment. In addition, this system allows for the quantification of tumor cell invasion by calculating distances that invaded cells move from the primary tumor site. Overall this procedure provides an enhanced model system for analyzing factors that contribute to SCOT invasion and therapeutic treatments tailored to prevent local invasion and distant metastatic spread. This method also has the potential to be ultimately combined with other imaging modalities in an in vivo setting.

A comprehensive overview of the techniques involved in generating a mouse model of oral cancer and quantitative monitoring of tumor invasion within the tongue through multi-photon microscopy of labeled cells is presented. This system can serve as a useful platform for the molecular assessment and drug efficacy of anti-invasive compounds.

Multi-Photonen-Imaging der Tumorzellinvasion in einem orthotopen Mausmodell des oralen Plattenepithelkarzinoms

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

Subsurface Defect Localization by Structured Heating Using Laser Projected Photothermal Thermography

The presented method is used to locate subsurface defects oriented perpendicularly to the surface. To achieve this, we create destructively interfering thermal wave fields that are disturbed by the defect. This effect is measured and used to locate the defect. We form the destructively interfering wave fields by using a modified projector. The original light engine of the projector is replaced with a fiber-coupled high-power diode laser. Its beam is shaped and aligned to the projector&#39;s spatial light modulator and optimized for optimal optical throughput and homogeneous projection by first characterizing the beam profile, and, second, correcting it mechanically and numerically. A high-performance infrared (IR) camera is set up according to the tight geometrical situation (including corrections of the geometrical image distortions) and the requirement to detect weak temperature oscillations at the sample surface. Data acquisition can be performed once a synchronization between the individual thermal wave field sources, the scanning stage, and the IR camera is established by using a dedicated experimental setup which needs to be tuned to the specific material being investigated. During data post-processing, the relevant information on the presence of a defect below the surface of the sample is extracted. It is retrieved from the oscillating part of the acquired thermal radiation coming from the so-called depletion line of the sample surface. The exact location of the defect is deduced from the analysis of the spatial-temporal shape of these oscillations in a final step. The method is reference-free and very sensitive to changes within the thermal wave field. So far, the method has been tested with steel samples but is applicable to different materials as well, in particular to temperature sensitive materials.

This method aims at locating vertical subsurface defects. Here, we couple a laser with a spatial light modulator and trigger its video input to heat a sample surface deterministically with two anti-phased modulated lines while acquiring highly resolved thermal images. The defect position is retrieved from evaluating thermal wave interference minima.

Untergrundfehler-Lokalisierung durch strukturierte Erwärmung mittels laserprojektierter photothermischer Thermografie

The presented method is used to locate subsurface defects oriented perpendicularly to the surface. To achieve this, we create destructively interfering ...

Ingenieurwesen

Universal Hand-held Three-dimensional Optoacoustic Imaging Probe for Deep Tissue Human Angiography and Functional Preclinical Studies in Real Time

The exclusive combination of high optical contrast and excellent spatial resolution makes optoacoustics (photoacoustics) ideal for simultaneously attaining anatomical, functional and molecular contrast in deep optically opaque tissues. While enormous potential has been recently demonstrated in the application of optoacoustics for small animal research, vast efforts have also been undertaken in translating this imaging technology into clinical practice. We present here a newly developed optoacoustic tomography approach capable of delivering high resolution and spectrally enriched volumetric images of tissue morphology and function in real time. A detailed description of the experimental protocol for operating with the imaging system in both hand-held and stationary modes is provided and showcased for different potential scenarios involving functional and molecular studies in murine models and humans. The possibility for real time visualization in three dimensions along with the versatile handheld design of the imaging probe make the newly developed approach unique among the pantheon of imaging modalities used in today&#8217;s preclinical research and clinical practice.

We provide herein a detailed description of the experimental protocol for imaging with a newly developed hand-held optoacoustic (photoacoustic) system for three-dimensional functional and molecular imaging in real time. The demonstrated powerful performance and versatility may define new application areas of the optoacoustic technology in preclinical research and clinical practice.

Universal-Hand Dreidimensionale Optoakustische Bildgebung Sonde für Deep Tissue Menschen Angiographie und Funktions präklinischen Studien in Echtzeit

The exclusive combination of high optical contrast and excellent spatial resolution makes optoacoustics (photoacoustics) ideal for simultaneously ...

Fabrication and Characterization of a Conformal Skin-like Electronic System for Quantitative, Cutaneous Wound Management

Recent advances in the development of electronic technologies and biomedical devices offer opportunities for non-invasive, quantitative assessment of cutaneous wound healing on the skin. Existing methods, however, still rely on visual inspections through various microscopic tools and devices that normally include high-cost, sophisticated systems and require well trained personnel for operation and data analysis. Here, we describe methods and protocols to fabricate a conformal, skin-like electronics system that enables conformal lamination to the skin surface near the wound tissues, which provides recording of high fidelity electrical signals such as skin temperature and thermal conductivity. The methods of device fabrication provide details of step-by-step preparation of the microelectronic system that is completely enclosed with elastomeric silicone materials to offer electrical isolation. The experimental study presents multifunctional, biocompatible, waterproof, reusable, and flexible/stretchable characteristics of the device for clinical applications. Protocols of clinical testing provide an overview and sequential process of cleaning, testing setup, system operation, and data acquisition with the skin-like electronics, gently mounted on hypersensitive, cutaneous wound and contralateral tissues on patients.

This article presents methods to fabricate and characterize a conformal, skin-like electronic system and protocols for the use in clinical applications, particularly on cutaneous wound management.

Herstellung und Charakterisierung eines Conformal-Haut-like Electronic System for Quantitative, kutane Wundmanagement

Recent advances in the development of electronic technologies and biomedical devices offer opportunities for non-invasive, quantitative assessment of ...

Combining Reflectance Confocal Microscopy with Optical Coherence Tomography for Noninvasive Diagnosis of Skin Cancers via Image Acquisition

Skin cancer is one of the most common cancers worldwide. Diagnosis relies on visual inspection and dermoscopy followed by biopsy for histopathological confirmation. While the sensitivity of dermoscopy is high, the lower specificity results in 70%-80% of the biopsies being diagnosed as benign lesions on histopathology (false positives on dermoscopy).
Reflectance confocal microscopy (RCM) and optical coherence tomography (OCT) imaging can noninvasively guide the diagnosis of skin cancers. RCM visualizes cellular morphology in en-face layers. It has doubled the diagnostic specificity for melanoma and pigmented keratinocytic skin cancers over dermoscopy, halving the number of biopsies of benign lesions. RCM acquired billing codes in the USA and is now being integrated into clinics.
However, limitations such as the shallow depth (~200 &#181;m) of imaging, poor contrast for nonpigmented skin lesions, and imaging in en-face layers result in relatively lower specificity for the detection of nonpigmented basal cell carcinoma (BCCs) &#8212; superficial BCCs contiguous with the basal cell layer and deeper infiltrative BCCs. In contrast, OCT lacks cellular resolution but images tissue in vertical planes down to a depth of ~1 mm, which allows the detection of both superficial and deeper subtypes of BCCs. Thus, both techniques are essentially complementary.
A &#34;multi-modal,&#34; combined RCM-OCT device simultaneously images skin lesions in both en-face and vertical modes. It is useful for the diagnosis and management of BCCs (nonsurgical treatment for superficial BCCs vs. surgical treatment for deeper lesions). A marked improvement in specificity is obtained for detecting small, nonpigmented BCCs over RCM alone. RCM and RCM-OCT devices are bringing a major paradigm shift in the diagnosis and management of skin cancers; however, their use is currently limited to academic tertiary care centers and some private clinics. This paper familiarizes clinicians with these devices and their applications, addressing translational barriers into routine clinical workflow.

Here, we describe protocols for acquiring good-quality images using novel, noninvasive imaging devices of reflectance confocal microscopy (RCM) and combined RCM and optical coherence tomography (OCT). We also familiarize clinicians with their clinical applications so that they can integrate the techniques into regular clinical workflows to improve patient care.

Kombination von konfokaler Reflexionsmikroskopie mit optischer Kohärenztomographie zur nicht-invasiven Diagnose von Hautkrebs mittels Bildaufnahme

Skin cancer is one of the most common cancers worldwide. Diagnosis relies on visual inspection and dermoscopy followed by biopsy for histopathological ...

Krebsforschung

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 ...

Intravital Microscopy of Tumor-associated Vasculature Using Advanced Dorsal Skinfold Window Chambers on Transgenic Fluorescent Mice

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static information and is often not sufficient for a correct interpretation. Intravital evaluation, in which a process is followed in time, provides extra and often unexpected information. With the creation of transgenic animals expressing cell-specific markers and live cell tracers, improvements to imaging equipment, and the development of several imaging chambers, intravital microscopy has become an important tool to better understand biological processes. This paper describes an experimental design for the investigation of tumor vessel development and of therapeutic effects in a spatial and temporal manner. Using this setup, the stage of vessel development, tip cell and lumen formation, blood flow, extravasation, an established vascular bed, and vascular destruction can be visualized and followed. Furthermore, therapeutic effects, intratumoral fate, and the localization of chemotherapeutic compounds can also be followed.

This protocol describes the intravital imaging of transgenic mice expressing cell-specific fluorescent markers. Intravital imaging provides a non-invasive method for high-resolution observations in living animals using implantable windows through which the microscopic visualization of processes is possible. This method is especially useful to study longitudinal processes.

Intravitalen Mikroskopie von Tumor-assoziierten Gefäßsystem mit erweiterten dorsalen Hautfalte Fenster Kammern an transgenen Mäusen fluoreszierende

Tumor and tumor vessel development, as well as tumor response to therapeutics, are highly dynamic biological processes. Histology provides static ...

Infrared Thermography for the Detection of Changes in Brown Adipose Tissue Activity

Measuring brown adipose tissue (BAT) activity by positron emission tomography computed tomography (PET-CT) via the accumulation of 18F-fluorodeoxyglucose (FDG) after a meal or in obese or diabetic patients fails as the method of choice. The main reason is that 18F-FDG competes with the postprandial high glucose plasma concentration for the same glucose transporter on the membrane of BAT cells. In addition, BAT uses fatty acids as a source of energy as well, which is not visible with PET-CT and could be changed along with glucose concentration in obese and diabetic patients. Therefore, to estimate the physiological importance of BAT in animals and humans, a new infrared thermography method used in recent publications is applied.
After overnight fasting, BAT activity was measured by infrared thermography before and after a meal in human volunteers and&#160;female wild-type mice. The camera software calculates the object&#39;s temperature using distance from the object, skin emissivity, reflected room temperature, air temperature, and relative humidity. In mice, the shaved area above the BAT was a region of interest for which average and maximal temperatures were measured. The phase of the estrous cycle in female mice was determined after an experiment by vaginal smears stained with cresyl violet (0.1%) stain solution. In healthy volunteers, two skin areas of the neck were selected: the supraclavicular area (above the collarbone, where BAT cells are present) and the interclavicular area (between the collarbones, where there is no BAT tissue detected). BAT activity is determined by the subtraction of those two values. Also, the average and maximal temperatures of skin areas could be determined in animals and human participants.
Changes in BAT activity after a meal measured by infrared thermography, a non-invasive and more sensitive method, were shown to be sex, age, and phase of the estrous cycle dependent in laboratory animals. As part of diet-induced thermogenesis, BAT activation in humans was also proven to be sex, age, and body mass index dependent. Further determining the pathophysiological changes in BAT activity after a meal will be of great importance for participants with high glucose plasma concentrations (obesity and diabetes mellitus type 2), as well as in different laboratory animals (knock-out mice). This method is also a variable tool for determining possible activating drugs that could rejuvenate BAT activity.

Here, we present a protocol for measuring brown adipose tissue activity after a meal in humans and laboratory animals.

Infrarot-Thermografie zur Detektion von Veränderungen der Aktivität des braunen Fettgewebes

Measuring brown adipose tissue (BAT) activity by positron emission tomography computed tomography (PET-CT) via the accumulation of 18F-fluorodeoxyglucose ...