KO é apoiado por uma bolsa da American Heart Association.

Antes de começar a Coração do adulto <ol><li> Recém preparar hemolinfa artificial (AH) solução contendo 108mm Na +, 5mM K +, 2mM de Ca 2 + 8mm, MgCl 2, 1 mM NaH 2 PO 4, 4 mM NaHCO 3, sacarose 10mM, trealose 5mM, HEPES 5mM (pH 7.1, todos os reagentes de Chemicals Sigma). A sacarose e trealose deve ser adicionado ao AH a partir de soluções estoque refrigerado imediatamente antes do uso, a fim de evitar a contaminação bacteriana. </li><li> AH trazer à temperatura ambiente e oxigenar a solução por via aérea borbulhando por pelo menos 15 min. </li><li> Puxe vários capilares finos (por exemplo, capilares de vidro, 100ul, VWR) necessário para a remoção de gorduras. </li></ol> Corações larval <ol><li> Sylgard revestido lamínulas: usando um palito de transferência, de cerca de 50-70μl de solução Sylgard em uma lamela 22x22mm. Deixe o endurecem Sylgard a 65 ° C durante a noite. </li><li> Prepare o Broadie e buffer de Bate (135mm NaCl, KCl 5mM, 4mm MgCl 2, CaCl 2 2mM, TES 5mM, Sacarose 36mm;. PH 7,15 Veja Broadie e Bate, 1993). </li><li> Prepare as ferramentas necessárias para aplicar a cola Histoacryl. Primeiro, puxe vários capilares finos (por exemplo, Ciência Produtos GB100T8P). Conecte uma ponta de pipeta de 1,5 ml com um pequeno tubo de plástico (Tygon, 1 / 32 &quot;), inserindo a extremidade menor para dentro do tubo. Inserir um capilar com a sua ampla abertura para o outro lado. Carregar o capilar com cola Histoacryl por imersão do capilar em uma gota de cola e uma sucção suave na ponta de plástico. Ao trabalhar com a cola deve ter um segundo copo capilar em mãos para remover a cola excessiva da ponta da cola capilar. A cola é líquido até que ele entra em contato com solução salina, onde ele irá endurecer em poucos segundos. No entanto, ele geralmente permanece plástica por tempo suficiente para ser removido da ponta cola capilar ou, aplicando-o ao modelo. </li></ol> Semi-intactas Coração Drosophila da mosca adulta <ol><li> Moscas Drosophila são anestesiados com Nap Fly (Carolina Biological Supply Co.) por 2-5 minutos. É preciso ter cuidado para não deixar as moscas no Nap Fly para exposições mais longas. Exposição a curto prazo ao frio também pode ser usado para anestesiar as moscas, mas CO 2 não é recomendado, pois tem mais efeitos duradouros sobre a freqüência cardíaca e ritmo. </li><li> Anestesiados moscas são colocadas, lado dorsal para baixo, em uma placa de Petri revestida com uma fina camada de vaselina. A cutícula hidrofóbica nas asas e no corpo reversivelmente &quot;pau&quot; para a geléia, mas a mosca pode ser reposicionada, se necessário. Isso se torna especialmente importante durante as manipulações subsequentes. </li><li> Um corte inicial é feita usando um par de tesouras curvas mola (Roboz # 5611). As lâminas de tesoura são colocados sob as pernas e anguladas para baixo em direção a superfície dorsal do tórax perto do pescoço. A cabeça, cordão nervoso ventral, e as pernas são removidos com um único corte. A preparação é então imerso em um hemolinfa, oxigenado artificial (AH) solução. </li><li> Usando a tesoura primavera a ponta posterior do abdômen (composto por segmentos abdominais 7 e 8) é removido com um único corte. Isso fornece acesso para fazer cortes laterais ao longo de ambas bordas do abdome e serve para cortar a ligação entre o intestino posterior e da cutícula abdominal. O retalho livre de cutícula abdominal ventral é então removido. </li><li> Nos passos 3 e 4 as conexões anterior e posterior do intestino são cortados para os órgãos abdominais são agora mantidos no lugar apenas por tracheols. O intestino e outros órgãos abdominais geralmente pode ser removida como uma única massa com a pinça joalheiro (Roboz, # 55) e suave puxando. Remover os órgãos internos revela o tubo coração batendo ainda ligado à cutícula dorsal cercado por corpos de gordura. </li><li> Os corpos de gordura são bastante opacos no microscópio de luz, consequentemente, muitas vezes é necessário remover algumas dessa gordura, a fim de ver o tubo cardíaco claramente. Isto é realizado por uma técnica de lipoaspiração usando finamente desenhadas capilares de vidro. Capilares são feitas usando um padrão pipeta extrator (Eg Sutter P-97 Pipetar Puller) e depois são inseridos em tubos de plástico (Tygon, 1 / 16 &quot;) ligado a uma fonte de vácuo. Este sistema é usado para sugar o excesso de gordura ao redor do tubo de coração. Força de sucção é proporcional ao diâmetro da ponta para pipetas com pontas maiores que 40 mícrons de diâmetro não deve ser usado. Extremo cuidado deve ser tomado para evitar tocar o próprio coração e porque a parte posterior do coração é muito frágil naquela região deve ser evitada. </li><li> O tubo de coração adulto Drosophila está agora exposta e deve ser de bater. Se necessário, o coração cutícula e anexado pode ser reposicionada, delicadamente levantar a cutícula fora da vaselina e cuidadosamente tamping-lo de volta para baixo, novamente evitando qualquer contato com o tecido do coração. Em tseu ponto de vista a solução que banha a preparação deve ser substituído por AHL fresco. Esta preparação deve ser autorizada a estabilizar durante 20-30 minutos com oxigenação antes de qualquer manipulação. Se a preparação deve ser mantida por mais de 60 minutos deve ser perfundido com AHL fresco. </li></ol> Imobilizando larvas Drosophila para gravação óptica <ol><li> Coloque uma gota de cola Histoacryl perto de cada canto do lamínulas Sylgard. Adicionar 50μl de tampão B &amp; B para as gotas de cola que vai endurecer instantaneamente. Preencher o espaço entre os pontos colados com B &amp; B - a cola vai manter o buffer no slide. </li><li> Dê uma vagando larva L3 e colocá-lo em um pedaço de lenço de papel molhado em gelo por 2 min. Isto irá desacelerar seu movimento. Em seguida, transferir a larva no buffer e orientá-la com o lado dorsal para cima (este é o lado da larva que não tem os cintos dentículo). Aplicar rapidamente um pouco de cola entre a região da cabeça e do Sylgard. A cola vai endurecer muito rapidamente e impedir que a cabeça da larva de se mover. </li><li> Utilizando uma pinça cuidadosamente agarrar o animal pela espiráculos posterior e gentilmente esticá-lo. Fixar a larva nesta posição, aplicando a cola ao longo de ambos os lados da larva na metade posterior. Para reduzir ainda mais o movimento da cutícula pelos músculos da parede do corpo, adicione mais cola para os flancos do animal. A larva é agora fixa nesta posição. A lamela pode ser colocado em uma placa de Petri pequena cheia de AHL ou PBS. Batimentos cardíacos podem ser gravados sem a interferência de movimentos parede do corpo. </li></ol> Resultados representativos: A semi-intactas adulto coração vai bater ritmicamente Drosophila por horas após a dissecação, quando mantidos em fresco, AH oxigenado (ver Dados Suplementar, Ocorr, et al., 2007). Um exemplo representativo é mostrado no filme 1. Corações de 3 º instar larva e moscas jovens (1-3 semanas após a eclosão) geralmente apresentam um batimento cardíaco regular em taxas entre 1-3 Hz. Voa com mais de três semanas, geralmente são mais arrítmicos, no entanto, esses corações ainda são capazes de bater espontaneamente por horas em AHL oxigenado. Corações em moscas adultas que estão danificados, como resultado do procedimento de dissecção normalmente apresentam regiões localizadas de constrição extremas que são incapazes de relaxar. Taxas de batimento cardíaco mais lento do que 1 Hz são raramente observadas em moscas com menos de 3 semanas de idade, conseqüentemente tais preparações são consideradas danificadas e são descartados. <a href="http://www.jove.com/files/ftp_upload/1425/MOVIE1.avi">Clique aqui para baixar Filme 1.</a> - A uma semana fly tipo selvagem de idade adulta (w cepa de laboratório 1118) mostrando o tubo de coração exposto abdominal (anterior à esquerda). Nota regular, contrações rítmicas. Regiões opacas no canto superior direito estão os restantes órgãos de gordura abdominal; células redondas de cada lado do coração são as células do pericárdio.

<ol><li>Ocorr, K., Reeves, N., Wessells, R. J., Fink, M., Chen, H. -S. V., Akasaka, T., Yasuda, S., Metzger, J., Giles, W., Posakony, J. W., Bodmer, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((KCNQ+potassium+channel+mutations+cause+cardiac+arrhythmias+in+Drosophila+that+mimic+the+effects+of+aging%5BTitle%5D)+AND+%22Proc+Natl+Acad+Sci+U+S+A%22%5BJournal%5D)">KCNQ potassium channel mutations cause cardiac arrhythmias in Drosophila that mimic the effects of aging.</a> Proc Natl Acad Sci U S A. 104, 3943-3948 (2007).</li>
<li>Broadie, K. S., Bate, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/pubmed?term=((Development+of+the+embryonic+neuromuscular+synapse+of+Drosophila+melanogaster%5BTitle%5D)+AND+%22J+Neurosci%22%5BJournal%5D)">Development of the embryonic neuromuscular synapse of Drosophila melanogaster.</a> J Neurosci. 13, 144-166 (1993).</li>
</ol>

O modelo de Drosophila provou ser uma poderosa ferramenta genética que tem sido utilizado para tratar uma variedade de questões científicas que vão desde desenvolvimento embrionário para a aprendizagem e memória. Recentemente, este organismo modelo versátil, tem sido usado para investigar a genética da função cardíaca na mosca. Uma série de tentativas para quantificar a fisiologia do coração em adultos Drosophila têm contado com observações feitas em moscas intacta através da cutícula abdominal. A mai...

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visualizing the beating heart in drosophila

O<em&gt; Drosophila</emCardíaca&gt; surgiu recentemente como um sistema bom modelo para examinar os mecanismos genéticos, celulares e moleculares subjacentes à função em corações miogênica. Um passo fundamental na análise da função cardíaca na mosca é encontrar uma maneira de acessar o coração de uma forma que preserva a sua função miogênica enquanto ainda permitindo que o órgão coração batendo a serem observados e registrados. Dois métodos diferentes para observar e registrar o coração batendo em ambos larva e adulto<em&gt; Drosophila</em&gt; São descritos aqui. Nossa preparação semi-intactas usando moscas adultas permite a visualização clara do coração abdominal, sem interferência da cutícula pigmentadas e sobrejacente corpos gordos. Para gravar coração bate larval é necessário imobilizar a larva, o que minimiza os movimentos do corpo, reduzindo assim os movimentos da parede do coração que não estão associados com as contrações do miocárdio. Nossas metodologias produzir adulto estável e preparações de coração larval que pode bater por horas a taxas de Hz 1-3.

Watch this Scientific Journal Video about Visualizing the Beating Heart in Drosophila at JoVE.com

Visualizing the Beating Heart in Drosophila

Técnica necessária para visualizar o coração batendo em larvas e adultos<em&gt; Drosophila</em&gt; São apresentados. Cada fase da vida exige uma metodologia diferente.

Visualizar o coração batendo no Drosophila</em

The Drosophila heart has recently emerged as a good model system for examining the genetic, cellular, and molecular mechanisms underlying function in ...

visualizing-the-beating-heart-in-drosophila

Research

JoVE Journal

Biology

Visualizing the Beating Heart in Drosophila

18.7K visualizações.  The Sanford Burnham Institute for Medical Research. Técnica necessária para visualizar o coração batendo em larvas e adultos Drosophila São apresentados. Cada fase da vida exige uma metodologia diferente.

Vídeo: Visualizing the Beating Heart in Drosophila

Visualizing the Live Drosophila Glial-neuromuscular Junction with Fluorescent Dyes

Our project identified GFP labeled glial structures at the developing larval fly neuromuscular synapse. To look at development of live glial-nerve-muscle synapses, we developed a larval tissue preparation that had features of live intact larvae, but also had good optical properties.  This new preparation also allowed for access of perfusates to the synapse.  We used fly larvae, immersed them in artificial hemolymph, and relaxed their normal rhythmic body contractions by chilling them. Next we dissected off the posterior segments of each animal and with a blunt insect pin pushed the mouth parts backward through the body cavity.  This everted the larval body wall, like turning a sock inside-out.  We completed the dissection with ultra-fine dissection scissors and thus exposed the visceral side of the body wall muscles. The glial structures at the NMJ expressed membrane targeted GFP under the control of glial specific promoters. The post-synaptic membrane, the SSR (Subsynaptic Reticula) in muscle expressed synaptically targeted dsRed. We needed to acutely label the motor neuron terminals, the third part of the synapse. To do this we applied primary antibodies to HRP, conjugated to a far-red emitting flurophore.  To test for dye diffusion properties into the perisynaptic space between the motor neuron terminals and the SSR, we applied a solution of large Dextran molecules conjugated to far-red emitting flurophore and collected images.

Visualizing the Live Drosophila Glial-neuromuscular Junction with Fluorescent Dyes

We described structural features of the Glia-neuromuscular synapses in a novel Inside-out tissue preparation of live fly larvae using fluorescent dyes with confocal microscopy. We labeled live neuron terminals with fluorescent primary antibodies to HRP, and also visualized the perisynaptic space with fluorescent Dextrans.

Visualizando o Live Drosophila Junção neuromuscular-Glial com corantes fluorescentes

Our project identified GFP labeled glial structures at the developing larval fly neuromuscular synapse. To look at development of live glial-nerve-muscle ...

Biologia

Fluorescent Labeling of Drosophila Heart Structures

The Drosophila melanogaster dorsal vessel, or heart, is a tubular structure comprised of a single layer of contractile cardiomyocytes, pericardial cells that align along each side of the heart wall, supportive alary muscles and, in adults, a layer of ventral longitudinal muscle cells. The contractile fibers house conserved constituents of the muscle cytoarchitecture including densely packed bundles of myofibrils and cytoskeletal/submembranous protein complexes, which interact with homologous components of the extracellular matrix. Here we describe a protocol for the fixation and the fluorescent labeling of particular myocardial elements from the hearts of dissected larvae and semi-intact adult Drosophila. Specifically, we demonstrate the labeling of sarcomeric F-actin and of &alpha;-actinin in larval hearts. Additionally, we perform labeling of F-actin and &alpha;-actinin in myosin-GFP expressing adult flies and of &alpha;-actinin and pericardin, a type IV extracellular matrix collagen, in wild type adult hearts. Particular attention is given to a mounting strategy for semi-intact adult hearts that minimizes handling and optimizes the opportunity for maintaining the integrity of the cardiac tubes and the associated tissues. These preparations are suitable for imaging via fluorescent and confocal microscopy. Overall, this procedure allows for careful and detailed analysis of the structural characteristics of the heart from a powerful genetically tractable model system.

Fluorescent Labeling of Drosophila Heart Structures

Here we describe a basic protocol for fluorescent labeling of different elements of heart tubes from larva and adult Drosophila melanogaster. These specimens are well-suited for imaging via fluorescent or confocal microscopy. This technique permits detailed structural analysis of the features of the hearts from a powerful model organism.

Labeling fluorescentes de Drosophila Estruturas Coração

The Drosophila melanogaster dorsal vessel, or heart, is a tubular structure comprised of a single layer of contractile cardiomyocytes, pericardial cells ...

Semi-automated Optical Heartbeat Analysis of Small Hearts

We have developed a method for analyzing high speed optical recordings from Drosophila, zebrafish and embryonic mouse hearts (Fink, et. al., 2009). Our Semi-automatic Optical Heartbeat Analysis (SOHA) uses a novel movement detection algorithm that is able to detect cardiac movements associated with individual contractile and relaxation events. The program provides a host of physiologically relevant readouts including systolic and diastolic intervals, heart rate, as well as qualitative and quantitative measures of heartbeat arrhythmicity. The program also calculates heart diameter measurements during both diastole and systole from which fractional shortening and fractional area changes are calculated. Output is provided as a digital file compatible with most spreadsheet programs. Measurements are made for every heartbeat in a record increasing the statistical power of the output. We demonstrate each of the steps where user input is required and show the application of our methodology to the analysis of heart function in all three genetically tractable heart models.

We have developed a Semi-automated Optical Heartbeat Analysis method (SOHA) for analyzing high speed optical recordings from Drosophila, zebrafish and embryonic mouse hearts. We demonstrate the application of our methodology to the analysis of heart function in fruit fly and embryonic mouse hearts.

Semi-automático de análise de pulsação óptica de pequenos corações

We have developed a method for analyzing high speed optical recordings from Drosophila, zebrafish and embryonic mouse hearts (Fink, et. al., 2009). Our ...

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

We present various methods to record cardiac function in the larval Drosophila. The approaches allow heart rate to be measured in unrestrained and restrained whole larvae. For direct control of the environment around the heart another approach utilizes the dissected larvae and removal of the internal organs in order to bathe the heart in desired compounds. The exposed heart also allows membrane potentials to be monitored which can give insight of the ionic currents generated by the myocytes and for electrical conduction along the heart tube. These approaches have various advantages and disadvantages for future experiments that are discussed. The larval heart preparation provides an additional model besides the Drosophila skeletal NMJ to investigate the role of intracellular calcium regulation on cellular function. Learning more about the underlying ionic currents that shape the action potentials in myocytes in various species, one can hope to get a handle on the known ionic dysfunctions associated to specific genes responsible for various diseases in mammals.

Monitoring Heart Function in Larval Drosophila melanogaster for Physiological Studies

We present various ways to monitor heart function in the larva of Drosophila for assessing questions dealing with the function of gap junctions, ion channel mutations, modulation of pacemaker activity and pharmacological studies.

Função de monitorização cardíaca em larval Drosophila melanogaster Para estudos fisiológicos

We present various methods to record cardiac function in the larval Drosophila. The approaches allow heart rate to be measured in unrestrained  and  ...

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

The morphogenesis of the Drosophila embryonic heart tube has emerged as a valuable model system for studying cell migration, cell-cell adhesion and cell shape changes during embryonic development. One of the challenges faced in studying this structure is that the lumen of the heart tube, as well as the membrane features that are crucial to heart tube formation, are difficult to visualize in whole mount embryos, due to the small size of the heart tube and intra-lumenal space relative to the embryo. The use of transmission electron microscopy allows for higher magnification of these structures and gives the advantage of examining the embryos in cross section, which easily reveals the size and shape of the lumen. In this video, we detail the process for reliable fixation, embedding, and sectioning of late stage Drosophila embryos in order to visualize the heart tube lumen as well as important cellular structures including cell-cell junctions and the basement membrane.

Preparation of embryos for Electron Microscopy of the Drosophila embryonic heart tube

We describe a process for fixation, embedding, sectioning, and imaging of late stage Drosophila embryos for Trasmission Electron Microscopy of the embryonic heart tube. This technique allows for the visualization of the heart tube lumen as well as the basement membrane, which lines the lumen of the heart.

Preparação de embriões de Microscopia Eletrônica do Drosophila Tubo cardíaco embrionário

The morphogenesis of the Drosophila  embryonic heart tube has emerged as a valuable model system for studying cell migration, cell-cell adhesion and cell ...

Novel And Efficient Method for Drosophila Heart Fluorescence Staining with Cryosectioning

Drosophila heart models are widely employed in studying cardiac aging and modeling&#160;human cardiac diseases. However, the dissection of Drosophila hearts before imaging is a meticulous time-intensive process that requires advanced training and motor skills. To address these challenges, we present an innovative protocol that utilizes cryosectioning for the fluorescence imaging of Drosophila heart tissue. The protocol has been demonstrated in imaging the adult Drosophila heart but could be adapted for developmental stages. The method enhances both the efficiency and accessibility of fluorescence staining while preserving the integrity of the tissue. This protocol simplifies the process without compromising the quality of imaging, thereby reducing the dependency on technicians with highly developed training and motor skills. Specifically, we replace complex techniques, such as capillary vacuum suction, with more straightforward methods like tissue embedding. This approach allows for the visualization of cardiac structures with greater ease and reproducibility. We demonstrate the utility of this protocol by effectively detecting key cardiac markers and&#160;achieving high-resolution fluorescence and immunostaining imaging that unveils intricate details of heart morphology and cellular organization. This method provides a robust and accessible tool for researchers exploring Drosophila cardiac biology, facilitating detailed analyses of heart development, function, and disease models.

The Drosophila heart sectioning and fluorescence imaging protocol simplifies studying heart structure and pathologies. This approach involves straightforward sectioning, staining, and imaging, bypassing the technical expertise needed for traditional dissection. It enhances accessibility, making Drosophila a more widely usable model for cardiac-related research within the broader scientific community.

Método novo e eficiente para coloração de fluorescência cardíaca de Drosophila com crioseccionamento

Drosophila heart models are widely employed in studying cardiac aging and modeling&#160;human cardiac diseases. However, the dissection of Drosophila ...

Genética

Developing Drosophila melanogaster Models for Imaging and Optogenetic Control of Cardiac Function

Using Drosophila melanogaster (fruit fly) as a model organism has ensured significant progress in many areas of biological science, from cellular organization and genomic investigations to behavioral studies. Due to the accumulated scientific knowledge, in recent years, Drosophila was brought to the field of modeling human diseases, including heart disorders. The presented work describes the experimental system for monitoring and manipulating the heart function in the context of a whole live organism using red light (617 nm) and without invasive procedures. Control over the heart was achieved using optogenetic tools. Optogenetics combines the expression of light-sensitive transgenic opsins and their optical activation to regulate the biological tissue of interest. In this work, a custom integrated optical coherence tomography (OCT) imaging and optogenetic stimulation system was used to visualize and modulate the functioning D. melanogaster heart at the 3rd instar larval and early pupal developmental stages. The UAS/GAL4 dual genetic system was employed to express halorhodopsin (eNpHR2.0) and red-shifted channelrhodopsin (ReaChR), specifically in the fly heart. Details on preparing D. melanogaster for live OCT imaging and optogenetic pacing are provided. A lab-developed integration software processed the imaging data to create visual presentations and quantitative characteristics of Drosophila heart function. The results demonstrate the feasibility of initiating cardiac arrest and bradycardia caused by eNpHR2.0 activation and performing heart pacing upon ReaChR activation.

Developing Drosophila melanogaster Models for Imaging and Optogenetic Control of Cardiac Function

The present protocol describes the generation of Drosophila melanogaster expressing eNpHR2.0 or ReaChR opsins in the heart for OCT imaging and optogenetic heart pacing. Detailed instructions for Drosophila OCT imaging and heart beating modulation, including the simulation of restorable heart arrest, bradycardia, and tachycardia in live animals at different developmental stages, are reported.

Desenvolvendo Modelos de Melanogaster Drosophila para Imagem e Controle Optogenético da Função Cardíaca

Using Drosophila melanogaster (fruit fly) as a model organism has ensured significant progress in many areas of biological science, from cellular ...

Bioengenharia

Drosophila Preparation and Longitudinal Imaging of Heart Function In Vivo Using Optical Coherence Microscopy (OCM)

Longitudinal study of the heartbeat in small animals contributes to understanding structural and functional changes during heart development. Optical coherence microscopy (OCM) has been demonstrated to be capable of imaging small animal hearts with high spatial resolution and ultrahigh imaging speed. The high image contrast and noninvasive properties make OCM ideal for performing longitudinal studies without requiring tissue dissections or staining. Drosophila has been widely used as a model organism in cardiac developmental studies due to its high number of orthologous human disease genes, its similarity of molecular mechanisms and genetic pathways with vertebrates, its short life cycle, and its low culture cost. Here, the experimental protocols are described for the preparation of Drosophila and optical imaging of the heartbeat with a custom OCM system throughout the life cycle of the specimen. By following the steps provided in this report, transverse M-mode and 3D OCM images can be acquired to conduct longitudinal studies of the Drosophila cardiac morphology and function. The en face and axial sectional OCM images and the heart rate (HR) and cardiac activity period (CAP) histograms, were also shown to analyze the heart structural changes and to quantify the heart dynamics during Drosophila metamorphosis, combined with the videos constructed with M-mode images to trace cardiac activity intuitively. Due to the genetic similarity between Drosophila and vertebrates, longitudinal study of heart morphology and dynamics in fruit flies could help reveal the origins of human heart diseases. The protocol here would provide an effective method to perform a wide range of studies to understand the mechanisms of cardiac diseases in humans.

Drosophila Preparation and Longitudinal Imaging of Heart Function In Vivo Using Optical Coherence Microscopy OCM

Here, the experimental protocols are described for preparing Drosophila at different developmental stages and performing longitudinal optical imaging of Drosophila heartbeats using a custom optical coherence microscopy (OCM) system. The cardiac morphological and dynamical changes can be quantitatively characterized by analyzing the heart structural and functional parameters from OCM images.

Drosophila Preparação e imagem longitudinal do coração Função In Vivo Usando Optical Coherence Microscopy (OCM)

Longitudinal study of the heartbeat in small animals contributes to understanding structural and functional changes during heart development. Optical ...

Light Sheet Microscopy of Fast Cardiac Dynamics in Zebrafish Embryos

Embryonic cardiac research has greatly benefited from advances in fast in vivo light sheet fluorescence microscopy (LSFM). Combined with the rapid external development, tractable genetics, and translucency of the zebrafish, Danio rerio, LSFM has delivered insights into cardiac form and function at high spatial and temporal resolution without significant phototoxicity or photobleaching. Imaging of beating hearts challenges existing sample preparation and microscopy techniques. One needs to maintain a healthy sample in a constricted field of view and acquire ultrafast images to resolve the heartbeat. Here we describe optimized tools and solutions to study the zebrafish heart in vivo. We demonstrate the applications of bright transgenic lines for labeling the cardiac constituents and present novel gentle embedding and immobilization techniques that avoid developmental defects and changes in heart rate. We also propose a data acquisition and analysis pipeline adapted to cardiac imaging. The entire workflow presented here focuses on zebrafish embryonic heart imaging but can also be applied to various other samples and experiments.

We describe optimized tools to study the zebrafish heart in vivo with light sheet fluorescence microscopy. Specifically, we suggest bright cardiac transgenic lines and present new gentle embedding and immobilization techniques that avoid developmental and heart defects. A possible data acquisition and analysis pipeline adapted to cardiac imaging is also provided.

Microscopia de Folha de Luz da Dinâmica Cardíaca Rápida em Embriões de Zebrafish

Embryonic cardiac research has greatly benefited from advances in fast in vivo light sheet fluorescence microscopy (LSFM). Combined with the rapid ...

Preparation of Drosophila Heart: A Technique to Study Heart Physiology in the Adult Fly

Source: Vogler, G., Ocorr, K. <a href="https://www.jove.com/video/1425/" target="_blank">Visualizing the Beating Heart in Drosophila</a>. J. Vis. Exp. (2009).
This video describes the fly&#39;s adult heart, or otherwise called the Drosophila dorsal vessel. The featured protocol demonstrates a dissection procedure for live heart preparations.

Visualizar o coração batendo no Drosophila

Resumo

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