The research leading to these results has received funding from the European Research Council under grant agreement ERC-2010-StG-260991.

Hacimsel elle tutulan optoacoustic probu ile çalıştırmak için ayrıntılı bir prosedür aşağıda tarif edilmiştir. Bu prosedür hayvan ve insan deneyleri ile ilgili onaylanan kurumsal düzenlemelere göre yapılır. 1. Sistem Hazırlama <ol><li> Çıkış ışık demeti stabilize Ameliyat öncesinde 15 dk&#39;lık bir ısınma dönemi için lazer açın. </li><li> Derinin (Şekil 1) ile temas halinde olan izole membran ile su kapama parçası yerleştirin. Not: (deri ile temas halinde olan) ayırma membranı ile dönüştürücüye (küresel prob merkezi) maksimum hassasiyeti bölgesi arasındaki mesafe etkin görüntüleme derinliğini oluşturur. </li><li> Bir pompa vasıtası ile izole membran ve deiyonize su ile transdüktörün yüzeyi arasında yaklaşık olarak 100 ml tam hacmma doldurun. </li><li> Herhangi bir su kaçağı veya hava kabarcıkları prese olduğundan emin olunnt. Seçenek olarak ise, su sirkülasyon sağlayarak hava kabarcıklarını önlemek. </li><li> Oda sıcaklığında deneyleri gerçekleştirmek ve bu sıcaklıkta muhafaza edilir, bağlantı ortamı (su) sunuyor. </li></ol> 2. Görüntüleme Hazırlık <ol><li> İnsan görüntüleme hazırlanması. </li><li> Görüntülerde istenmeyen bir arka plan önlemek için bir epilasyon losyon ile to-görüntülü parçası saç kaldırmak (bu adım isteğe bağlıdır). </li><li> Bölgesi çevresinde cilt açıdan verimli akustik bağlantı mekanizmasının temin etmek amacıyla, görüntülenecek üzerine ultrason jeli uygulanır. Ilgi bölgesinde optoacoustic probu yerleştirin. Hiçbir hava kabarcığı ultrason bağlantı jel mevcut olduğundan emin olun. </li><li> Hayvan görüntüleme hazırlanması. </li><li> Hayvanlar ile bakım ve deneysel işlemler, kurumsal ve devlet kuralları ve düzenlemeleri ile uyum içinde olduğundan emin olun. </li><li> Bölgesinde hayvanın kürk çıkarın bir traş losyon ile, görüntülenecek. Ile hayvanın gözleri koruyunyoğun darbeli lazer radyasyonuna maruz kuruluk ve hasar görmesini önler veteriner merhem. </li><li> 0.9 l, ketamin / ksilazin hacim olarak (100 mg / kg ketamin + 5 mg / kg iksilazin) Deneyden önce ya da kullanım izofluran anestezisi (% 2-3 () intraperitoneal (İP) kullanarak, hayvan anestezisi / Deney sırasında dakika gaz akışı). Hayvanın arka bacak refleksini kontrol ederek anestezisi teyit edin. </li><li> Bölgenin etrafında cilde ultrason jeli uygulayın verimli akustik bağlamayı sağlayan ve ilgi bölgesinde optoacoustic probu yerleştirmek için yansıması için. Hiçbir hava kabarcığı ultrason bağlantı jel mevcut olduğundan emin olun. </li></ol> 3. Ön görünümü Çalışma Modu <ol><li> 690 nm ve 900 nm ve 10 ile 50 Hz arasında darbe tekrarlama oranı arasındaki görüntüleme dalga boyu (ler) kurma. Akustik veri toplama sistemi için parametreleri seçin - 1 MΩgiriş empedansı. İkinci ve 12 bit dikey çözünürlük başına 40 megasamples bir örnekleme hızında her lazer darbesi için 2030 örnekleri Edinme. Lazer Q-switch çıkışı ile satın tetikler. </li><li> Operatör ve hasta hem de optik dalgaboyu (ler) için uyarlanmış koruyucu gözlük kullanın emin olun. Insan deneyleri 19 emniyet maruz kalma limitlerini karşılamak ve hayvanlarda termal baskıyı ve cilt hasarı önlemek için doku yüzeyindeki ışık fluens yakın kızılötesi dalga boyları için deney sırasında 20 mJ / cm 2 altında tutulur, böylece lazer gücünü ayarlama . </li><li> Lazer darbe tekrarlama oranı karşılık gelen bir kare hızında üç boyutlu görüntülemeye izin işleme algoritmalarının bir GPU uygulaması ile ön görünümü yazılımını başlatın. </li><li> Prob Taşı ve / veya nesne görselleştirme performansını optimize etmek ve ilgi yapılarını lokalize etmek için görüntülü edilecek. </li></ol> 4. Veri Toplama <ol><li> Tarama (el) modu için veri toplama. </li><li> Gerekirse, ilgi bölgesinde kontrast zenginleştirmek alımı öncesinde bir kontrast madde enjekte. NOT: Bizim deneylerde kontrastlı insan görüntüleme gerçekleştirilen değil. Bununla birlikte, çeşitli kontrast maddeler, potansiyel olarak, bu amaç için kullanılabilir. Indosiyanin yeşili (ICG) yetişkin vücut ağırlığı 2 mg / kg&#39;lık maksimum tavsiye edilen dozda kontrast için kullanılabilir klinik olarak onaylı bir optik kontrast maddesinin bir örnektir. </li><li> Önizleme yazılımın yürütülmesini sağlamak 3.1 anlatılan parametreler ile, veri toplama için uygun donanım başlatın. Yavaşça ilgi yapılarını izlemek için görüntülü bölgenin etrafında probu hareket. NOT: Birden fazla lazer dalga boylarında görüntüler eşzamanlı olarak yakalandığı zaman, el modunda prob hareket hızı önemli ölçüde azalma olduğu (tercihen aşağıda 2 mm / snIşıksal karışmamış görüntülerde hareket ilgili eserler önlemek için 50 Hz lazer darbe tekrarlama oranı) için. </li><li> Sabit modu için veri toplama. </li><li> Tutucuya görüntülü nesne (örn., Hayvan) ve el probu Dağı ve ön görünümü yazılımın yürütülmesini sağlamak 3.1 açıklanan parametreleri ile alımı başlatabilirsiniz. </li><li> Optoacoustic prob ve ilgi bölgesi dinamik biyolojik olayların görselleştirmek için deney boyunca aynı pozisyonda görüntüleme parçası koruyun. </li><li> Ilgi bölgedeki dinamik dağılımını izlemek için kontrast madde enjekte edilir. NOT: Bizim fare deneylerinde, Indosiyanin Yeşil (ICG) kontrast için kullanılmıştır. Genel bir kılavuz, 10 nmol ve 0.4 mg bir miktarda itibariyle / ICG&#39;nin kg, in vivo olarak, multi-spektral optoacoustics ile tespit kontrast oluşturmak için farenin dolaşımı içine alınmak üzere yer alır. NOT: kontrast madde olmalıdırilgili makam tarafından, insan ve / veya hayvan kullanımı için onaylanmıştır. </li></ol> 5. Deney sonlandırılması <ol><li> Lazer durdurun. </li><li> Görüntülü bölgeden optoacoustic probu çıkarın. Hayvan çalışmada, anestezi kaynağını durdurun. </li><li> Sıcak tutmak ve tam anestezi kurtarıldı kadar diğer hayvanlarla temasını önlemek için bir kızılötesi ısıtıcı altında hayvan yerleştirin. Anestezi kurtarma sırasında gözetimsiz hayvan bırakmayın. </li></ol> 6. Off-line Veri İşleme <ol><li> Veri işleme için kullanılan yazılım uygulamasında edinilen optoacoustic sinyalleri içeren dosya (lar) yükleyin. </li><li> Her bir çerçeve ve her dalga boyu için optik absorpsiyon volümetrik bir görüntüye karşılık gelen üç boyutlu bir matris dizi elde etmek üzere bir rekonstrüksiyon algoritması kullanarak. NOT: bu tercih edilir rekonstrüksiyonu için böyle Heterojen gibi bozan faktörler, muhasebe bir algoritma kullanmakgörüntüsü nesne de çaba ve zayıflama sırayla nihai bant genişliği ve bulgulama elemanları ve hafif akıcılık varyasyonlar geometrik şekil, etkisi, soğurulan enerjinin dağılımının daha nicel bir gösterimini elde etmek için. </li><li> Her bir çoklu dalga boyu çerçeve, numunede mevcut olan her bir emici madde için optik absorpsiyon temsil eden üç boyutlu bir matris dizileri yeni bir dizi elde etmek için bir karışmama algoritması kullanılır. </li><li> Gerekirse, daha fazla biyolojik ilgili parametrelerin görüntülenmesine ve okumayı kolaylaştırmak için optik soğurma dağılımını temsil eden matris dizileri işlemek. </li></ol>

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The authors have nothing to disclose.

Küçük hayvan araştırmalarında optoacoustic görüntüleme teknikleri ile sunulan benzersiz avantajlar örneğin öngörülen teşhis ve tedavi izleme uygulamaları bir dizi ile, klinik uygulamaya teknolojiyi tercüme için güçlü motivasyon yarattı., Meme ve cilt kanseri, iltihap veya periferik damar hastalıkları. Bununla birlikte, etkili bir tomografik görüntü elde etme geometrisini oluşturmak için ışık kaynakları ve tespit elemanlarının yeterli sayıda çevrili olabilir fareler veya daha...

Teknolojinin 1-5 sunduğu eşsiz avantajlar sömürmek yeni uygulamaların çeşitliliği kapsayan yayınların giderek artan sayısına göre tecelli olarak Optoacoustic (foto akustik) görüntüleme, biyolojik ve tıbbi araştırma topluluklarından büyüyen ilgi çekiyor. Özellikle, uzak ışık uzadıya sınırının ötesinde derinliklerde yüksek uzay-zamansal çözünürlüğe sahip görüntü spektral ayırt edici foto-emici ajanlara kapasite fonksiyonel ve moleküler görüntüleme 6-10 görülmemiş yetenekleri açılır. Nitekim, klinik uygulamaya optoacoustic teknolojinin çeviri teşhis ve birçok hastalığın tedavi takibinde umut verici umutları ile birlikte geliyor. Bununla birlikte, optik olarak saçılma fotonlar ve emici doku ve optoacoustic olgusu ile ilişkili olarak çok zayıf tepkiler sınırlı yayılma yöntemi uygulanabilir derinliğini sınırlar. Bunun bir sonucu olarak, elle tutulan optoaendoskopi sistemleri doğal menfeze 13 yoluyla onlara takarak vücutta görüntüleri sağlamak için kullanılır ise kostik sondalar vücut 11,12 dışından erişilebilir görüntü parçaları için denenmiştir. Böyle kadın meme gibi insan vücudunun bazı düşük emici parçalar, ayrıca tomografik optoacoustic tarayıcılar 14,15 ile erişilebilir. Ayni şekilde, ultrasonografi, büyük esnekliği sağladığından Özel ilgi konusu olan, elde tutulan bir yaklaşımdır. Burada, optoacoustic görüntüleme için ortak ultrason doğrusal dizi prob adaptasyon başlıca nedeni, ultrason ve optoacoustics arasında tomografik görüntüleme gereksinimleri temel farklılıklar nedeniyle, zorlu kalır. Standart ultrasonografide yüksek kare hızları sıralı olarak etkin iken kHz aralığında yüksek darbe tekrarlama frekansları kullanan şemaları gönderme-alma, gerçek zamanlı üç boyutlu optoacoustic görüntüleme tek bir int hacimsel tomografik eşzamanlı veri toplama ile elde edilirlazer nabzını errogating. Bu durumda, yüksek kaliteli optoacoustic görüntüleme görüntüsü nesnenin etrafında en olası katı açısı üç boyutlu verilerin elde edilmesi anlamına gelir. Son zamanlarda, biz gerçek zamanlı 16 üç boyutlu (hacimsel) görüntüleme için ilk el optoacoustic probu tanıttı. Sistem 90 ° &#39;lik bir açı içeren küresel bir yüzey (Şekil 1A&#39; da mavi noktalar) üzerine yerleştirilmiş 256 piezoelektrik elemanlarının bir iki boyutlu dizi dayanmaktadır. Yaklaşık 3 x 3 mm 2&#39;nin tek tek elemanlarının boyutu, aynı zamanda bunların oryantasyon ve frekans bant genişliği (yaklaşık olarak 2-6 MHz) Şekil kürenin merkezini (siyah küp saran bir santimetre çaplı bir birimden etkili sinyal toplama garanti 1A). Görüntüleme bölgesinde optik uyarım, dizinin merkezi bir silindirik boşluğu içine sokulan bir elyaf demeti ile donatılır, böylece herhangi bir dalga boyu suscElyaf demeti aracılığıyla iletilen edilme eptible görüntüleme için kullanılabilir. Optik fiber demeti ile transdüktörün dizinin bir gerçek resmi Şekil 1B &#39;de gösterilmiştir. Lazer darbe tekrarlama frekansı tarafından belirlenen bir kare hızında gerçek zamanlı görüntüleme ayrıca bir Grafikler- ile etkin şekilde verimli uyarma ve sinyallerin eş zamanlı algılama, tek atış uyarma (bir lazer darbesi) ile derin doku görüntüleme sağlar işlem birimi (GPU) rekonstrüksiyon prosedürü 17. uygulanması. Saydam polietilen membran (Şekil 1C) ile silindirik bir mahfaza, bir akustik verici sıvı ortam (su) çevrelemek için dönüştürücü dizisine bağlanmıştır. Zar, başka akustik jel aracılığıyla dokuya bağlanır. Elle tutulan bir işletim modunda kullanılır bir şekilde optoacoustic probunun bir resmi Şekil 1D &#39;de gösterilmiştir. Gösterdi thrklinik teşhis ve potansiyel uygulamalar bir dizi için önemli avantajlar ile gelen gerçek zamanlı fonksiyonel görüntüleme kapasitesi ile birlikte ee boyutlu el optoacoustic görüntüleme, periferik vasküler hastalık gibi çeşitli endikasyonlar, lenfatik sistem bozuklukları, meme kanseri, deri lezyonları için öngörülmektedir enflamasyon veya artrit 18. Ayrıca, hızlı görüntüleme kapasitesi sabit bir konumda düzenlenmiş probu ile dinamik bir biyolojik olayların görselleştirme sağlar. Hızlı dalga boyu ayarlama optik parametrik osilatör (AFE) lazer teknolojisi ile birleştiğinde, bu yaklaşım foto-emici maddeler biodistribution gerçek zamanlı görüntüleme için izin verir. Böylece, yeni olanaklar aynı derecede örneğin, küçük hayvan görüntüleme uygulamalarında ortaya çıkabilir., Doku hemodinamiğini okuyan, in vivo hücre izleme, farmakokinetik, organ perfüzyon görselleştirme, tümörler ve kalp-damar sistemi, veya nörogörüntülemenin moleküler görüntüleme hedefli. <p class = biz birkaç tipik klinik ve küçük hayvan görüntüleme senaryolarda küresel dizi optoacoustic el prob ve vitrin performans ile çalışması için deneysel görüntüleme protokolü ayrıntılı bir açıklamasını bu çalışmada &quot;jove_content&quot;&gt;.

<table border="0" cellpadding="0" cellspacing="0" width="1663">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Name of Material/ Equipment</td>
			<td style="width:389px;">Company</td>
			<td style="width:171px;">Catalog Number</td>
			<td style="width:777px;">Comments/Description</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:327px;">Optical parametric oscillator (OPO)-based laser</td>
			<td style="width:389px;">Innolas Laser GmbH, Krailling (Germany)</td>
			<td style="width:171px;">custom-made</td>
			<td style="width:777px;">The laser provides laser pulses with a duration around 10ns and an energy up to 80mJ. 
			The wavelength is tunable between 680-950nm.</td>
		</tr>
		<tr height="45">
			<td height="45" style="height:45px;width:327px;">Spherical array of piezocomposite detectors</td>
			<td style="width:389px;">Imasonic SaS, Voray (France)</td>
			<td style="width:171px;">custom-made</td>
			<td style="width:777px;">The array consists of 256 piezoelectric sensors distributed on a spherical surface. 
			Each element has dimensions 3x3mm2, a central frequency of 4MHz and a bandwidth of 100%.&#160;</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:327px;">Data acquisition system (DAQ)</td>
			<td style="width:389px;">Falkenstein Mikrosysteme GmbH, Taufkirchen (Germany)</td>
			<td style="width:171px;">custom-made</td>
			<td style="width:777px;">The DAQ simultaneously acquires 256 signals at 40 megasamples per second and 2030 samples. 
			The input impedance is 1MW.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:327px;">Fiber bundle</td>
			<td style="width:389px;">CeramOptec GmbH, Bonn (Germany)</td>
			<td style="width:171px;">custom-made</td>
			<td style="width:777px;">The bundle consists of 480 individual fibers randomly distributed in the input and output. 
			The numerical aperture of each individual fiber is 0.22.</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:327px;">Athymic Nude mouse</td>
			<td style="width:389px;">Harlan Laboratories (The Netherlands)</td>
			<td style="width:171px;">Athymic nude - Foxn1nu</td>
			<td style="width:777px;">The mouse was 8 weeks old (adult) at the time of the experiment. 
			The ethical protocol was approved by the Bavarian goverment (number 55.2.1.54-2632-102-11)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:327px;">Bepanthen cream</td>
			<td style="width:389px;">Bayer AG (Germany)</td>
			<td style="width:171px;">N/A</td>
			<td>Vet ointment to protect the eyes during anesthesia</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:327px;">Data processing software</td>
			<td style="width:389px;">Matlab (Mathworks, Natick, MA, USA)</td>
			<td style="width:171px;">custom-made</td>
			<td style="width:777px;">The data processing software was devoped at our institute. 
			It allows reconstruction at each wavelength and multi-wavelength unmixing, as well as further data processing.</td>
		</tr>
		<tr height="24">
			<td height="24" style="height:24px;width:327px;">Water-enclosing part</td>
			<td style="width:389px;">N/A</td>
			<td style="width:171px;">custom-made</td>
			<td>This part contains the water that acts as an acoustic coupling medium between skin and transducer elements</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:327px;">Indocyanine green (ICG)</td>
			<td style="width:389px;">PULSION Medical Systems SE</td>
			<td style="width:171px;">N/A</td>
			<td style="width:777px;">ICG-PULSION (active ingredient: indocyanine green dye) is a drug used in cardiac, circulatory and micro-circulatory diagnostics, liver function diagnostics and ophthalmic angiography diagnostics.</td>
		</tr>
	</tbody>
</table>

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.

Açıklanan hacimsel el optoacoustic prob yeteneklerini gösteren Temsilcisi sonuçları, bu bölümde sergilendi. Her durumda, deri yüzeyi üzerinde ışık akıcılığı 20 mJ / cm2 19 emniyet maruz kalma sınırının altında tutuldu. Periferik insan damarsal izleme gerçek zamanlı prob performansı Şekil 2 teşhir edilmiştir. Bu deney süresince, prob yavaş yavaş lazer çalışma ile 800 nm tek bir dalga boyunda sağlıklı bir insan gönüllü elin bo...

Watch this Scientific Journal Video about Universal Hand-held Three-dimensional Optoacoustic Imaging Probe for Deep Tissue Human Angiography and Functional Preclinical Studies in Real Time at JoVE.com

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

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.

Evrensel Real Time Derin Doku İnsan Anjiografi ve Fonksiyonel Preklinik Çalışmalar üç boyutlu Optoacoustic Görüntüleme Probe El

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

universal-hand-held-three-dimensional-optoacoustic-imaging-probe-for

Research

JoVE Journal

Bioengineering

10.6K Görüntüleme.  Helmholtz Zentrum München. 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.

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

Three-dimensional Optical-resolution Photoacoustic Microscopy

Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As the mainstays of in vivo three-dimensional (3-D) optical microscopy, single-/multi-photon fluorescence microscopy and optical coherence tomography (OCT) have demonstrated their extraordinary sensitivities to fluorescence and optical scattering contrasts, respectively. However, the optical absorption contrast of biological tissues, which encodes essential physiological/pathological information, has not yet been assessable. 
The emergence of biomedical photoacoustics has led to a new branch of optical microscopy optical-resolution photoacoustic microscopy (OR-PAM)1, where the optical irradiation is focused to the diffraction limit to achieve cellular1 or even subcellular2 level lateral resolution. As a valuable complement to existing optical microscopy technologies, OR-PAM brings in at least two novelties. First and most importantly, OR-PAM detects optical absorption contrasts with extraordinary sensitivity (i.e., 100%). Combining OR-PAM with fluorescence microscopy3 or with optical-scattering-based OCT4 (or with both) provides comprehensive optical properties of biological tissues. Second, OR-PAM encodes optical absorption into acoustic waves, in contrast to the pure optical processes in fluorescence microscopy and OCT, and provides background-free detection. The acoustic detection in OR-PAM mitigates the impacts of optical scattering on signal degradation and naturally eliminates possible interferences (i.e., crosstalks) between excitation and detection, which is a common problem in fluorescence microscopy due to the overlap between the excitation and fluorescence spectra. 
Unique for optical absorption imaging, OR-PAM has demonstrated broad biomedical applications since its invention, including, but not limited to, neurology5, 6, ophthalmology7, 8, vascular biology9, and dermatology10. In this video, we teach the system configuration and alignment of OR-PAM as well as the experimental procedures for in vivo functional microvascular imaging.

Optical-resolution photoacoustic microscopy (OR-PAM) is an emerging technology capable of imaging optical absorption contrasts in vivo with cellular resolution and sensitivity. Here, we provide a visualized instruction on the experimental protocols of OR-PAM, including system configuration, system alignment, typical in vivo experimental procedures, and functional imaging schemes.

Üç boyutlu optik çözünürlük foto akustik Mikroskopi

Optical microscopy, providing valuable insights at the cellular and organelle levels, has been widely recognized as an enabling biomedical technology. As ...

Biyomühendislik

Planar and Three-Dimensional Printing of Conductive Inks

Printed electronics rely on low-cost, large-area fabrication routes to create flexible or multidimensional electronic, optoelectronic, and biomedical devices1-3. In this paper, we focus on one- (1D), two- (2D), and three-dimensional (3D) printing of conductive metallic inks in the form of flexible, stretchable, and spanning microelectrodes.
Direct-write assembly4,5 is a 1-to-3D printing technique that enables the fabrication of features ranging from simple lines to complex structures by the deposition of concentrated inks through fine nozzles (~0.1 - 250 &mu;m). This printing method consists of a computer-controlled 3-axis translation stage, an ink reservoir and nozzle, and 10x telescopic lens for visualization. Unlike inkjet printing, a droplet-based process, direct-write assembly involves the extrusion of ink filaments either in- or out-of-plane. The printed filaments typically conform to the nozzle size. Hence, microscale features (&lt; 1 &mu;m) can be patterned and assembled into larger arrays and multidimensional architectures.
In this paper, we first synthesize a highly concentrated silver nanoparticle ink for planar and 3D printing via direct-write assembly. Next, a standard protocol for printing microelectrodes in multidimensional motifs is demonstrated. Finally, applications of printed microelectrodes for electrically small antennas, solar cells, and light-emitting diodes are highlighted.

Planar and three-dimensional printing of conductive metallic inks is described. Our approach provides new avenues for fabricating printed electronic, optoelectronic, and biomedical devices in unusual layouts at the microscale.

İletken Mürekkepler düzlemsel ve Üç Boyutlu Baskı

Printed electronics rely on low-cost, large-area fabrication routes to create flexible or multidimensional electronic, optoelectronic, and biomedical ...

A Novel Three-dimensional Flow Chamber Device to Study Chemokine-directed Extravasation of Cells Circulating under Physiological Flow Conditions

Extravasation of circulating cells from the bloodstream plays a central role in many physiological and pathophysiological processes, including stem cell homing and tumor metastasis. The three-dimensional flow chamber device (hereafter the 3D device) is a novel in vitro technology that recreates physiological shear stress and allows each step of the cell extravasation cascade to be quantified. The 3D device consists of an upper compartment in which the cells of interest circulate under shear stress, and a lower compartment of static wells that contain the chemoattractants of interest. The two compartments are separated by porous inserts coated with a monolayer of endothelial cells (EC). An optional second insert with microenvironmental cells of interest can be placed immediately beneath the EC layer. A gas exchange unit allows the optimal CO2 tension to be maintained and provides an access point to add or withdraw cells or compounds during the experiment. The test cells circulate in the upper compartment at the desired shear stress (flow rate) controlled by a peristaltic pump. At the end of the experiment, the circulating and migrated cells are collected for further analyses. The 3D device can be used to examine cell rolling on and adhesion to EC under shear stress, transmigration in response to chemokine gradients, resistance to shear stress, cluster formation, and cell survival. In addition, the optional second insert allows the effects of crosstalk between EC and microenvironmental cells to be examined. The translational applications of the 3D device include testing of drug candidates that target cell migration and predicting the in vivo behavior of cells after intravenous injection. Thus, the novel 3D device is a versatile and inexpensive tool to study the molecular mechanisms that mediate cellular extravasation.

The three-dimensional flow chamber device is a novel in vitro technology for the quantitative and step-wise evaluation of the extravasation cascade of cells circulating under conditions of physiological shear stress. The device therefore fills a critical need for basic, preclinical, and clinical studies of cell migration.

Fizyolojik Akış Koşullarında Sirkülasyon Hücre Kemokin-yönettiği Ekstravazasyon Eğitim Bir Roman Üç boyutlu Akış Odası Cihazı

Extravasation of circulating cells from the bloodstream plays a central role in many physiological and pathophysiological processes, including stem cell ...

Human Cartilage Tissue Fabrication Using Three-dimensional Inkjet Printing Technology

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and regenerative medicine. With digital control&#160;cells, scaffolds, and growth factors can be precisely deposited to the desired two-dimensional (2D) and three-dimensional (3D) locations rapidly. Therefore, this technology is an ideal approach to fabricate tissues mimicking their native anatomic structures. In order to engineer cartilage with native zonal organization, extracellular matrix composition (ECM), and mechanical properties, we developed a bioprinting platform using a commercial inkjet printer with simultaneous photopolymerization capable for 3D cartilage tissue engineering. Human chondrocytes suspended in poly(ethylene glycol) diacrylate (PEGDA) were printed for 3D neocartilage construction via layer-by-layer assembly. The printed cells were fixed at their original deposited positions, supported by the surrounding scaffold in simultaneous photopolymerization. The mechanical properties of the printed tissue were similar to the native cartilage. Compared to conventional tissue fabrication, which requires longer UV exposure, the viability of the printed cells with simultaneous photopolymerization was significantly higher. Printed neocartilage demonstrated excellent glycosaminoglycan (GAG) and collagen type II production, which was consistent with gene expression. Therefore, this platform is ideal for accurate cell distribution and arrangement for anatomic tissue engineering.

The methods described in this paper show how to convert a commercial inkjet printer into a bioprinter with simultaneous UV polymerization. The printer is capable of constructing 3D tissue structure with cells and biomaterials. The study demonstrated here constructed a 3D neocartilage.

Üç boyutlu Mürekkep Püskürtmeli Baskı Teknolojisi Kullanımı İnsan Kıkırdak Doku Fabrikasyon

Bioprinting, which is based on thermal inkjet printing, is one of the most attractive enabling technologies in the field of tissue engineering and ...

Generation of a Three-dimensional Full Thickness Skin Equivalent and Automated Wounding

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models reflect the human wound situation better than animal models. Although two-dimensional models are widely used to investigate processes such as cellular migration and proliferation, models that are more complex are required to gain a deeper knowledge about wound healing. Besides a suitable model system, the generation of precise and reproducible wounds is crucial to ensure comparable results between different test runs. In this study, the generation of a three-dimensional full thickness skin equivalent to study wound healing is shown. The dermal part of the models is comprised of human dermal fibroblast embedded in a rat-tail collagen type I hydrogel. Following the inoculation with human epidermal keratinocytes and consequent culture at the air-liquid interface, a multilayered epidermis is formed on top of the models. To study the wound healing process, we additionally developed an automated wounding device, which generates standardized wounds in a sterile atmosphere.

The goal of this protocol is to build up a three-dimensional full thickness skin equivalent, which resembles natural skin. With a specifically constructed automated wounding device, precise and reproducible wounds can be generated under maintenance of sterility.

Üç-boyutlu Tam Kalınlığı Deri Üretimi Eşdeğer ve Otomatik Yaralama

In vitro models are a cost effective and ethical alternative to study cutaneous wound healing processes. Moreover, by using human cells, these models ...

Development of a Multicellular Three-dimensional Organotypic Model of the Human Intestinal Mucosa Grown Under Microgravity

Because cells growing in a three-dimensional (3-D) environment have the potential to bridge many gaps of cell cultivation in 2-D environments (e.g., flasks or dishes). In fact, it is widely recognized that cells grown in flasks or dishes tend to de-differentiate and lose specialized features of the tissues from which they were derived. Currently, there are mainly two types of 3-D culture systems where the cells are seeded into scaffolds mimicking the native extracellular matrix (ECM): (a) static models and (b) models using bioreactors. The first breakthrough was the static 3-D models. 3-D models using bioreactors such as the rotating-wall-vessel (RWV) bioreactors are a more recent development. The original concept of the RWV bioreactors was developed at NASA&#39;s Johnson Space Center in the early 1990s and is believed to overcome the limitations of static models such as the development of hypoxic, necrotic cores. The RWV bioreactors might circumvent this problem by providing fluid dynamics that allow the efficient diffusion of nutrients and oxygen. These bioreactors consist of a rotator base that serves to support and rotate two different formats of culture vessels that differ by their aeration source type: (1) Slow Turning Lateral Vessels (STLVs) with a co-axial oxygenator in the center, or (2) High Aspect Ratio Vessels (HARVs) with oxygenation via a flat, silicone rubber gas transfer membrane. These vessels allow efficient gas transfer while avoiding bubble formation and consequent turbulence. These conditions result in laminar flow and minimal shear force that models reduced gravity (microgravity) inside the culture vessel. Here we describe the development of a multicellular 3-D organotypic model of the human intestinal mucosa composed of an intestinal epithelial cell line and primary human lymphocytes, endothelial cells and fibroblasts cultured under microgravity provided by the RWV bioreactor.

Cells growing in a three-dimensional (3-D) environment represent a marked improvement over cell cultivation in 2-D environments (e.g., flasks or dishes). Here we describe the development of a multicellular 3-D organotypic model of the human intestinal mucosa cultured under microgravity provided by rotating-wall-vessel (RWV) bioreactors.

İnsan bağırsak mukozasının bir Multisellüler Üç boyutlu Organotipik Modelinin Geliştirilmesi yerçekimi Yetiştirilen

Because cells growing in a three-dimensional (3-D) environment have the potential to bridge many gaps of cell cultivation in 2-D environments (e.g., ...

Three-dimensional Tissue Engineered Aligned Astrocyte Networks to Recapitulate Developmental Mechanisms and Facilitate Nervous System Regeneration

Neurotrauma and neurodegenerative disease often result in lasting neurological deficits due to the limited capacity of the central nervous system (CNS) to replace lost neurons and regenerate axonal pathways. However, during nervous system development, neuronal migration and axonal extension often occur along pathways formed by other cells, referred to as &#34;living scaffolds&#34;. Seeking to emulate these mechanisms and to design a strategy that circumvents the inhibitory environment of the CNS, this manuscript presents a protocol to fabricate tissue engineered astrocyte-based &#34;living scaffolds&#34;. To create these constructs, we employed a novel biomaterial encasement scheme to induce astrocytes to self-assemble into dense three-dimensional bundles of bipolar longitudinally-aligned somata and processes. First, hollow hydrogel micro-columns were assembled, and the inner lumen was coated with collagen extracellular-matrix. Dissociated cerebral cortical astrocytes were then delivered into the lumen of the cylindrical micro-column and, at a critical inner diameter of &#60;350 &#181;m, spontaneously self-aligned and contracted to produce long fiber-like cables consisting of dense bundles of astrocyte processes and collagen fibrils measuring &#60;150 &#181;m in diameter yet extending several cm in length. These engineered living scaffolds exhibited &#62;97% cell viability and were virtually exclusively comprised of astrocytes expressing a combination of the intermediate filament proteins glial-fibrillary acidic protein (GFAP), vimentin, and nestin. These aligned astrocyte networks were found to provide a permissive substrate for neuronal attachment and aligned neurite extension. Moreover, these constructs maintain integrity and alignment when extracted from the hydrogel encasement, making them suitable for CNS implantation. These preformed constructs structurally emulate key cytoarchitectural elements of naturally occurring glial-based &#34;living scaffolds&#34; in vivo. As such, these engineered living scaffolds may serve as test-beds to study neurodevelopmental mechanisms in vitro or facilitate neuroregeneration by directing neuronal migration and/or axonal pathfinding following CNS degeneration in vivo.

We showcase the development of self-assembled, three-dimensional bundles of longitudinally aligned astrocytic somata and processes within a novel biomaterial encasement. These engineered &#34;living scaffolds&#34;, exhibiting micron-scale diameter yet extending centimeters in length, may serve as test-beds to study neurodevelopmental mechanisms or facilitate neuroregeneration by directing neuronal migration and/or axonal pathfinding.

Astrocyte ağlar gelişimsel mekanizmaları özetlemek ve sinir sistemi yeniden oluşturma işlemi kolaylaştırmak için tasarlanmış üç boyutlu doku uyumlu

Neurotrauma and neurodegenerative disease often result in lasting neurological deficits due to the limited capacity of the central nervous system (CNS) to ...

Melt Electrospinning Writing of Three-dimensional Poly(&#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology with great potential for biomedical applications. The technique facilitates the direct deposition of biocompatible polymer fibers to fabricate well-ordered scaffolds in the sub-micron to micro scale range. The establishment of a stable, viscoelastic, polymer jet between a spinneret and a collector is achieved using an applied voltage and can be direct-written. A significant benefit of a typical porous scaffold is a high surface-to-volume ratio which provides increased effective adhesion sites for cell attachment and growth. Controlling the printing process by fine-tuning the system parameters enables high reproducibility in the quality of the printed scaffolds. It also provides a flexible manufacturing platform for users to tailor the morphological structures of the scaffolds to their specific requirements. For this purpose, we present a protocol to obtain different fiber diameters using melt electrospinning writing (MEW) with a guided amendment of the parameters, including flow rate, voltage and collection speed. Furthermore, we demonstrate how to optimize the jet, discuss often experienced technical challenges, explain troubleshooting techniques and showcase a wide range of printable scaffold architectures.

Melt Electrospinning Writing of Three-dimensional Poly(&amp;#949;-caprolactone) Scaffolds with Controllable Morphologies for Tissue Engineering Applications

This protocol serves as a comprehensive guideline to fabricate scaffolds via electrospinning with polymer melts in a direct writing mode. We systematically outline the process and define the appropriate parameter settings for achieving targeted scaffold architectures.

Üç boyutlu Poly(ε-caprolactone) İskele kontrol edilebilir türleri morfoloji ile Electrospinning yazma doku mühendisliği uygulamaları için erime

This tutorial reflects on the fundamental principles and guidelines for electrospinning writing with polymer melts, an additive manufacturing technology ...

Hand-held Clinical Photoacoustic Imaging System for Real-time Non-invasive Small Animal Imaging

Translation of photoacoustic imaging into the clinic is a major challenge. Handheld real-time clinical photoacoustic imaging systems are very rare. Here, we report a combined photoacoustic and clinical ultrasound imaging system by integrating an ultrasound probe with light delivery for small animal imaging. We demonstrate this by showing sentinel lymph node imaging in small animals along with minimally invasive real-time needle guidance. A clinical ultrasound platform with access to raw channel data allows the integration of photoacoustic imaging leading to a handheld real-time clinical photoacoustic imaging system. Methylene blue was used for sentinel lymph node imaging at 675 nm wavelength. Additionally, needle guidance with dual modal ultrasound and photoacoustic imaging was shown using the imaging system. Depth imaging of up to 1.5 cm was demonstrated with a 10 Hz laser at a photoacoustic imaging frame rate of 5 frames per second.

A clinical handheld photoacoustic imaging system will be demonstrated for real-time non-invasive small animal imaging.

Görüntüleme sistemi gerçek zamanlı Non-invaziv küçük hayvan görüntüleme için el klinik Photoacoustic

Translation of photoacoustic imaging into the clinic is a major challenge. Handheld real-time clinical photoacoustic imaging systems are very rare. Here, ...

Culturing Mammalian Cells in Three-dimensional Peptide Scaffolds

A useful technique for culturing cells in a self-assembling nanofiber three-dimensional (3D) scaffold is described. This culture system recreates an environment that closely mimics the structural features of non-polarized tissue. Furthermore, the particular intrinsic nanofiber structure of the scaffold makes it transparent to visual light, which allows for easy visualization of the sample under microscopy. This advantage was largely used to study cell migration, organization, proliferation, and differentiation and thus any development of their particular cellular function by staining with specific dyes or probes. Furthermore, in this work, we describe the good performance of this system to easily study the redifferentiation of expanded human articular chondrocytes into cartilaginous tissue. Cells were encapsulated into self-assembling peptide scaffolds and cultured under specific conditions to promote chondrogenesis. Three-dimensional cultures showed good viability during the 4 weeks of the experiment. As expected, samples cultured with chondrogenic inducers (compared to non-induced controls) stained strongly positive for toluidine blue (which stains glycosaminoglycans (GAGs) that are highly present in cartilage extracellular matrix) and expressed specific molecular markers, including collagen type I, II and X, according to Western Blot analysis. This protocol is easy to perform and can be used at research laboratories, industries and for educational purposes in laboratory courses.

Here, we present a protocol to obtain 3D-culture systems in self-assembling peptide scaffolds to promote the differentiation of dedifferentiated human articular chondrocytes into cartilage-like tissue.

Üç boyutlu peptid iskele memeli hücrelerinde kültür

A useful technique for culturing cells in a self-assembling nanofiber three-dimensional (3D) scaffold is described. This culture system recreates an ...