O autor gostaria de reconhecer financiamento do NHLBI (K08 HL102066 e R01 HL114832).

<ol>
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The author has nothing to disclose.

Existem três passos críticos neste processo: 1) a colocação do tubo endotraqueal e ventilação adequado, 2) a colocação do cateter IV jugular, e 3) a colocação correcta do cateter PV no ventrículo esquerdo. Determinação da taxa respiratória apropriada é uma parte importante do fornecimento de suporte ventilatório. ratinhos conscientes geralmente manter a ventilação alveolar com respirações superficiais rápidas. Em geral, os ratinhos ventilados terá volumes correntes muito maiores. Assim, é necessá...

A doença cardiovascular continua a ser uma causa significativa de morbilidade e mortalidade em todo o mundo 1. Doenças do coração apresentar desafios particularmente difíceis no desenvolvimento de novas terapias. Os avanços no campo da genética para proporcionar a possibilidade de identificar uma grande variedade de potenciais contribuintes genética para o desenvolvimento de doença cardíaca. A natureza integradora do sistema cardiovascular exige que esses alvos genéticos ser validados em modelos animais intactos. Os custos de flexibilidade e baixa habitação genéticas do rato tê-lo trazido para o primeiro plano para a avaliação do papel fisiológico de um determinado gene. O pequeno tamanho do mouse apresenta alguns desafios únicos para a avaliação da função cardíaca. Existem várias modalidades que podem fornecer informações sobre a função cardíaca, mas apenas a medição simultânea da pressão ventricular e o volume permite pressão-volume (PV) análise do lacete da função ventricular. PV laços tudoOW função cardíaca a ser analisada independente da sua conexão com a vasculatura; um factor importante na determinação do papel funcional de um elemento genético particular. A avaliação de loops de pressão-volume tem sido utilizada tanto experimentalmente e clinicamente por muitos anos e extensa literatura existe sobre a análise desses conjuntos de dados 2,3. A adaptação da tecnologia de ciclo PV para o rato tem sido um importante avanço para a compreensão da fisiologia cardíaca murino 4-6. Cateter de tecnologias baseadas na ansa PV acoplar um transdutor de pressão e a utilização de condutância para estimar o volume do ventrículo. O volume ventricular é determinado por análise de alterações em um campo eléctrico gerado pelo cateter. Este método modelos do ventrículo como um cilindro, cuja altura é definida pela distância entre os eléctrodos no cateter e o raio é calculado a partir da condução de um campo eléctrico através do sangue emventrículo 7-9. O sinal de condutância medida pelo cateter tem duas componentes. A primeira é a condução através do sangue; esta varia com o volume do ventrículo e constitui o sinal primário usado para determinar o volume do ventrículo. O segundo componente resultante da condução através e ao longo da parede do ventrículo. Isto é chamado de condutância paralela e deve ser removido, a fim de determinar o volume do ventrículo absoluto. Há dois sistemas disponíveis comercialmente para a recolha de dados de pressão-volume no laboratório de pesquisa e o método utilizado para calcular e remover a condutância paralela é a principal diferença entre eles 6,10,11. cateteres condutância exigem a injecção de uma solução salina hipertónica para o cálculo da condutância paralelo. Esta injecção transitoriamente muda a condutividade do sangue no ventrículo, enquanto que a condutividade da parede mantém-se constante. A partir destes dados, é possível determinar acomponente do sinal de condutância que se origina a partir do sangue e o que vem a partir da parede do ventrículo. Esta abordagem assume que a condutância paralelo não varia durante o ciclo cardíaco. O método baseia-se na admissão mudanças de fase no campo eléctrico para avaliar a contribuição da parede ventricular para o sinal total de volume. Este método baseia-se uma variedade de constantes predeterminadas para a condutividade do sangue e do miocárdio para determinar o volume final, mas torna medidas contínuas de condutância paralela durante o ciclo cardíaco. Ambos estes sistemas fornecem boas estimativas do volume ventricular esquerdo e as diferenças entre eles não são susceptíveis de ser f isiologicamente significativa. O modelo cilíndrico do ventrículo e outras premissas tornar estas abordagens baseadas em cateter não tão precisos quanto outras modalidades, mas esses dados são fornecidos numa base de batimento a batimento que é essencial para a avaliação das medidas independentes de carga da função cardíaca. O procedimento aqui descrito é utilizado no meu laboratório e apresentou dados de um grande número de estudos que examinam os mecanismos patofisiológicos básicos de distrófica cardiomiopatia 12-18. O procedimento descrito abaixo é um dos dois que pode ser usado para obter dados de espira PV. Embora muitos dos princípios são aplicáveis ​​para qualquer abordagem, este protocolo vai se concentrar em uma abordagem apical de tórax aberto; um protocolo de tórax fechado foi detalhado em outros lugares 19,20. Enquanto o processo será descrito em pormenor, os princípios gerais são importantes para expor o coração com danos mínimos para quer o coração ou os pulmões. Durante todo o protocolo é importante lembrar que este é um procedimento não-sobrevivência e que tendo uma boa exposição do coração é criticamente importante para a colocação correcta do cateter.

<table border="0" cellpadding="0" cellspacing="0" width="523">
	<tbody>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Dumont 5/45 (2)</td>
			<td style="width:127px;">Fine Science Tools</td>
			<td style="width:119px;">11251-33</td>
			<td style="width:63px;"></td>
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			<td height="42" style="height:42px;width:216px;">Vessel Dilating Forceps</td>
			<td style="width:127px;">Fine Science Tools</td>
			<td style="width:119px;">18153-11</td>
			<td></td>
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			<td height="42" style="height:42px;width:216px;">Castroviejo Micro Dissecting Spring Scissor</td>
			<td style="width:127px;">Roboz Instruments</td>
			<td style="width:119px;">RS-5668</td>
			<td></td>
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			<td height="42" style="height:42px;width:216px;">Octogon Forceps - Serrated/Curved</td>
			<td style="width:127px;">Fine Science Tools</td>
			<td style="width:119px;">11041-08</td>
			<td></td>
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		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Octogon Forceps - Serrated/Straight</td>
			<td style="width:127px;">Fine Science Tools</td>
			<td style="width:119px;">11040-08</td>
			<td></td>
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			<td height="42" style="height:42px;width:216px;">Dissector Scissors- Heavy Blade</td>
			<td style="width:127px;">Fine Science Tools</td>
			<td style="width:119px;">14082-09</td>
			<td></td>
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			<td height="21" style="height:21px;width:216px;">Transpore Surgical Tape</td>
			<td style="width:127px;">3M</td>
			<td style="width:119px;">1527-1</td>
			<td></td>
		</tr>
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			<td height="42" style="height:42px;width:216px;">3-0 Silk Suture</td>
			<td style="width:127px;">Fine Science Tools</td>
			<td style="width:119px;">18020-30</td>
			<td></td>
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			<td height="21" style="height:21px;width:216px;">TOPO Ventilator</td>
			<td style="width:127px;">Kent Scientific</td>
			<td style="width:119px;">TOPO</td>
			<td></td>
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			<td height="42" style="height:42px;width:216px;">Martin ME 102 Electrosurgical Unit</td>
			<td style="width:127px;">Harvard Apparatus</td>
			<td style="width:119px;">PY2 72-2484</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Syringe Pump</td>
			<td style="width:127px;">Lucca Technologies</td>
			<td style="width:119px;">GenieTouch</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Stereomicroscope with boom stand</td>
			<td style="width:127px;">Nikon</td>
			<td style="width:119px;">SMZ-800N</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:216px;">Thermocouple Thermometer</td>
			<td style="width:127px;">Cole Parmer</td>
			<td style="width:119px;">EW-91100-40</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">T/Pump Warm Water Recirculator</td>
			<td style="width:127px;">Kent Scientific</td>
			<td style="width:119px;">TP-700</td>
			<td></td>
		</tr>
		<tr height="20">
			<td height="20" style="height:20px;width:216px;">ADVantage Pressure-Volume System</td>
			<td style="width:127px;">Transonic</td>
			<td style="width:119px;">ADV500</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:216px;">Data Acquision and Analysis</td>
			<td style="width:127px;">DSI</td>
			<td style="width:119px;">Ponemah ACQ-16</td>
			<td></td>
		</tr>
	</tbody>
</table>

measuring pressure volume loops in the mouse

Compreender as causas e progressão da doença cardíaca apresenta um desafio significativo para a comunidade biomédica. A flexibilidade genética do mouse oferece um grande potencial para explorar a função cardíaca no nível molecular. O tamanho reduzido do rato apresenta alguns desafios no que diz respeito à realização de fenotipagem cardíaco detalhado. Miniaturização e outros avanços na tecnologia fizeram muitos métodos de avaliação cardíaca possível no mouse. Destes, a coleta simultânea de dados de pressão e volume fornece uma imagem detalhada da função cardíaca que não está disponível através de qualquer outra modalidade. Aqui, um procedimento detalhado para a recolha de dados de loop pressão-volume é descrito. Incluído é uma discussão sobre os princípios subjacentes às medições e as potenciais fontes de erro. manejo anestésico e abordagens cirúrgicas são discutidos em grande detalhe como ambos são cruciais para a obtenção de medida hemodinâmica de alta qualidades. Os princípios do desenvolvimento protocolo de hemodinâmica e aspectos relevantes da análise dos dados também são abordados.

Por convenção, o volume é representada graficamente no eixo X e a pressão sobre o eixo dos Y como na Figura 1. As malhas de pressão-volume decorrentes traçando pressão contra o volume deve assemelhar a um rectângulo, os bordos verticais que representam as alterações isovolumétrica de pressão (isto é, quando ambos valvas mitral e aórtica estão fechados). A horizontal inferior representa enchimento ventricular através da válvula mitral e a parte h...

Watch this Scientific Journal Video about Measuring Pressure Volume Loops in the Mouse at JoVE.com

Measuring Pressure Volume Loops in the Mouse

Este manuscrito descreve um protocolo detalhado para a recolha de dados de pressão-volume do mouse.

Medição do volume Pressão Loops no mouse

Understanding the causes and progression of heart disease presents a significant challenge to the biomedical community. The genetic flexibility of the ...

measuring-pressure-volume-loops-in-the-mouse

Research

JoVE Journal

Biology

16.2K visualizações.  University of Minnesota.  Este manuscrito descreve um protocolo detalhado para a recolha de dados de pressão-volume do mouse. 

Vídeo: Measuring Pressure Volume Loops in the Mouse

Measuring Blood Pressure in Mice using Volume Pressure Recording, a Tail-cuff Method

The CODA 8-Channel High Throughput Non-Invasive Blood Pressure system measures the blood pressure in up to 8 mice or rats simultaneously. The CODA tail-cuff system uses Volume Pressure Recording (VPR) to measure the blood pressure by determining the tail blood volume. A specially designed differential pressure transducer and an occlusion tail-cuff measure the total blood volume in the tail without the need to obtain the individual pulse signal. Special attention is afforded to the length of the occlusion cuff in order to derive the most accurate blood pressure readings. VPR can easily obtain readings on dark-skinned rodents, such as C57BL6 mice and is MRI compatible. The CODA system provides you with measurements of six (6) different blood pressure parameters; systolic and diastolic blood pressure, heart rate, mean blood pressure, tail blood flow, and tail blood volume. Measurements can be made on either awake or anesthetized mice or rats. The CODA system includes a controller, laptop computer, software, cuffs, animal holders, infrared warming pads, and an infrared thermometer. There are seven different holder sizes for mice as small as 8 grams to rats as large as 900 grams.

The CODA 8-Channel High Throughput Non-Invasive Blood Pressure system measures the blood pressure in up to 8 mice or rats simultaneously. This tail-cuff system uses Volume Pressure Recording (VPR) to measure the blood pressure by determining the tail blood volume.

Medição da pressão arterial em ratos usando Recording Volume de pressão, um método de cauda-manguito

The CODA 8-Channel High Throughput Non-Invasive Blood Pressure system measures the blood pressure in up to 8 mice or rats simultaneously. The CODA ...

Biologia

Catheterization of Intestinal Loops in Ruminants

The intestine is a complex structure that is involved not only in absorption of nutrients, but also acts as a barrier between the individual and the outside world. As such, the intestine plays a pivotal role in immunosurveillance and protection from enteric pathogens. Investigating intestinal physiology and immunology commonly employs 'intestinal loops' as an experimental model. The majority of these loop models are non-recovery surgical procedures that study short-term (&lt;24 hr) changes in the intestine (1-3). We previously created a recovery intestinal loop model to specifically measure long-term (&lt;6 mo) immunological changes in the intestine of sheep following exposure to vaccines, adjuvants, and viruses (4). This procedure localized treatments to a specific 'loop', allowing us to sample this area of the intestine. A significant drawback of this method is the single window of opportunity to administer treatments (i.e. at the time of surgery). Furthermore, samples of both the intestinal mucosa and luminal contents can only be taken at the termination of the project. Other salient limitations of the above model are that the surgical manipulation and requisite post-operative measures (e.g. administration of antibiotics and analgesics) can directly affect the treatment itself and/or alter immune function, thereby confounding results. Therefore, we modified our intestinal loop model by inserting long-term catheters into the loops. Sheep recover fully from the procedure, and are unaffected by the exteriorized catheters. Notably, the establishment of catheters in loops allows us to introduce multiple treatments over an extended interval, following recovery from surgery and clearance of drugs administered during surgery and the post-operative period.

We describe a novel surgical method for catheterizing 'intestinal loops' within the ileum of sheep. Once animals have recovered from surgery and have cleared antibiotics and analgesics, multiple treatments can be deposited directly in loops via the catheters.

Cateterização de alças intestinais em Ruminantes

The intestine is a complex structure that is involved not only in absorption of nutrients, but also acts as a barrier between the individual and the ...

In vivo Imaging and Therapeutic Treatments in an Orthotopic Mouse Model of Ovarian Cancer

Human cancer and response to therapy is better represented in orthotopic animal models. This paper describes the development of an orthotopic mouse model of ovarian cancer, treatment of cancer via oral delivery of drugs, and monitoring of tumor cell behavior in response to drug treatment in real time using in vivo imaging system. In this orthotopic model, ovarian tumor cells expressing luciferase are applied topically by injecting them directly into the mouse bursa where each ovary is enclosed. Upon injection of D-luciferin, a substrate of firefly luciferase, luciferase-expressing cells generate bioluminescence signals. This signal is detected by the in vivo imaging system and allows for a non-invasive means of monitoring tumor growth, distribution, and regression in individual animals. Drug administration via oral gavage allows for a maximum dosing volume of 10 mL/kg body weight to be delivered directly to the stomach and closely resembles delivery of drugs in clinical treatments. Therefore, techniques described here, development of an orthotopic mouse model of ovarian cancer, oral delivery of drugs, and in vivo imaging, are useful for better understanding of human ovarian cancer and treatment and will improve targeting this disease.

In vivo Imaging and Therapeutic Treatments in an Orthotopic Mouse Model of Ovarian Cancer

Orthotopic animal models of ovarian cancer replicate better human disease and therefore enhance our understanding of cancer progression and tumor response to therapy. A mouse model receives an intrabursal injection of luciferase-expressing ovarian tumor cells. Treatment is administered via oral gavage. Tumor growth is monitored by in vivo imaging system.

In vivo de imagens e tratamentos terapêuticos em um modelo de camundongo ortotópico de câncer de ovário

Human cancer and response to therapy is better represented in orthotopic animal models.  This paper describes the development of an orthotopic mouse model ...

Measuring Caenorhabditis elegans Life Span in 96 Well Microtiter Plates

Lifespan is a biological process regulated by several genetic pathways. One strategy to investigate the biology of aging is to study animals that harbor mutations in components of age-regulatory pathways. If these mutations perturb the function of the age-regulatory pathway and therefore alter the lifespan of the entire organism, they provide important mechanistic insights1-3.
Another strategy to investigate the regulation of lifespan is to use small molecules to perturb age-regulatory pathways. To date, a number of molecules are known to extend lifespan in various model organisms and are used as tools to study the biology of aging4-16. The number of molecules identified thus far is small compared to the genetic "toolset" that is available to study the biology of aging.
Caenorhabditis elegans is one of the principle models used to study aging because of its excellent genetics and short lifespan of three weeks. More recently, C.elegans has emerged as a model organism for phenotype based drug screens5,7,16-20 because of its small size and its ability to grow in microtiter plates.
Here we present an assay to measure C.elegans lifespan in 96 well microtiter plates. The assay was developed and successfully used to screen large libraries for molecules that extend C.elegans lifespan7. The reliability of the assay was evaluated in multiple tests: first, by measuring the lifespan of wild type animals grown at different temperatures; second, by measuring the lifespan of mutants with altered lifespans; third, by measuring changes in lifespan in response to different concentrations of the antidepressant Mirtazepine. Mirtazepine has previously been shown to extend lifespan in C.elegans7. The results of these tests show that the assay is able to replicate previous findings from other assays and is quantitative. The microtiter format also makes this lifespan assay compatible with automated liquid handling systems and allows integration into automated platforms.

Measuring Caenorhabditis elegans Life Span in 96 Well Microtiter Plates

In this protocol we present a method to measure Caenorhabditis elegans lifespan in 96 well microtiter plates.

Medição Caenorhabditis elegans Expectativa de Vida em 96 placas de microtitulação Bem

Lifespan is a biological process regulated by several genetic pathways. One strategy to investigate the biology of aging is to study animals that harbor ...

Generation of Mice Derived from Induced Pluripotent Stem Cells

The production of induced pluripotent stem cells (iPSCs) from somatic cells provides a means to create valuable tools for basic research and may also produce a source of patient-matched cells for regenerative therapies. iPSCs may be generated using multiple protocols and derived from multiple cell sources. Once generated, iPSCs are tested using a variety of assays including immunostaining for pluripotency markers, generation of three germ layers in embryoid bodies and teratomas, comparisons of gene expression with embryonic stem cells (ESCs) and production of chimeric mice with or without germline contribution2. Importantly, iPSC lines that pass these tests still vary in their capacity to produce different differentiated cell types2. This has made it difficult to establish which iPSC derivation protocols, donor cell sources or selection methods are most useful for different applications.
The most stringent test of whether a stem cell line has sufficient developmental potential to generate all tissues required for survival of an organism (termed full pluripotency) is tetraploid embryo complementation (TEC)3-5. Technically, TEC involves electrofusion of two-cell embryos to generate tetraploid (4n) one-cell embryos that can be cultured in vitro to the blastocyst stage6. Diploid (2n) pluripotent stem cells (e.g. ESCs or iPSCs) are then injected into the blastocoel cavity of the tetraploid blastocyst and transferred to a recipient female for gestation (see Figure 1). The tetraploid component of the complemented embryo contributes almost exclusively to the extraembryonic tissues (placenta, yolk sac), whereas the diploid cells constitute the embryo proper, resulting in a fetus derived entirely from the injected stem cell line.
Recently, we reported the derivation of iPSC lines that reproducibly generate adult mice via TEC1. These iPSC lines give rise to viable pups with efficiencies of 5-13%, which is comparable to ESCs3,4,7 and higher than that reported for most other iPSC lines8-12. These reports show that direct reprogramming can produce fully pluripotent iPSCs that match ESCs in their developmental potential and efficiency of generating pups in TEC tests. At present, it is not clear what distinguishes between fully pluripotent iPSCs and less potent lines13-15. Nor is it clear which reprogramming methods will produce these lines with the highest efficiency. Here we describe one method that produces fully pluripotent iPSCs and "all- iPSC" mice, which may be helpful for investigators wishing to compare the pluripotency of iPSC lines or establish the equivalence of different reprogramming methods.

Generating induced pluripotent stem cell (iPSC) lines produces lines of differing developmental potential even when they pass standard tests for pluripotency. Here we describe a protocol to produce mice derived entirely from iPSCs, which defines the iPSC lines as possessing full pluripotency1.

Geração de camundongos derivadas de células-tronco pluripotentes induzidas

The production of induced pluripotent stem cells (iPSCs) from somatic cells provides a means to create valuable tools for basic research and may also ...

Measuring the Osmotic Water Permeability Coefficient (Pf) of Spherical Cells: Isolated Plant Protoplasts as an Example

Studying AQP regulation mechanisms is crucial for the understanding of water relations at both the cellular and the whole plant levels. Presented here is a simple and very efficient method for the determination of the osmotic water permeability coefficient (Pf) in plant protoplasts, applicable in principle also to other spherical cells such as frog oocytes. The first step of the assay is the isolation of protoplasts from the plant tissue of interest by enzymatic digestion into a chamber with an appropriate isotonic solution. The second step consists of an osmotic challenge assay: protoplasts immobilized on the bottom of the chamber are submitted to a constant perfusion starting with an isotonic solution and followed by a hypotonic solution. The cell swelling is video recorded. In the third step, the images are processed offline to yield volume changes, and the time course of the volume changes is correlated with the time course of the change in osmolarity of the chamber perfusion medium, using a curve fitting procedure written in Matlab (the &#8216;PfFit&#8217;), to yield Pf.

Measuring the Osmotic Water Permeability Coefficient Pf of Spherical Cells: Isolated Plant Protoplasts as an Example

Measuring the osmotic water permeability coefficient (Pf) of cells can help understand the regulatory mechanisms of aquaporins (AQPs). Pf determination in spherical plant cell protoplasts presented here involves protoplasts isolation and numerical analysis of their initial rate of volume change as a result of an osmotic challenge during constant bath perfusion.

Medindo o Coeficiente de Permeabilidade osmótica de água (P<sub&gt; F</sub&gt;) De células esféricas: protoplastos planta isolada como um exemplo

Studying AQP regulation mechanisms is crucial for the understanding of water relations at both the cellular and the whole plant levels. Presented here is ...

Measuring Glutathione-induced Feeding Response in Hydra

Hydra is among the most primitive organisms possessing a nervous system and chemosensation for detecting reduced glutathione (GSH) for capturing the prey. The movement of prey organisms causes mechanosensory discharge of the stinging cells called nematocysts from hydra, which are inserted into the prey. The feeding response in hydra, which includes curling of the tentacles to bring the prey towards the mouth, opening of the mouth and consequent engulfing of the prey, is triggered by GSH present in the fluid released from the injured prey. To be able to identify the molecular mechanism of the feeding response in hydra which is unknown to date, it is necessary to establish an assay to measure the feeding response. Here, we describe a simple method for the quantitation of the feeding response in which the distance between the apical end of the tentacle and mouth of hydra is measured and the ratio of such distance before and after the addition of GSH is determined. The ratio, called the relative tentacle spread, was found to give a measure of the feeding response. This assay was validated using a starvation model in which starved hydra show an enhanced feeding response in comparison with daily fed hydra.

Here we describe a simple assay for the quantification of the feeding response in hydra induced by the reduced form of glutathione. This assay relies on measuring the distance between the apical end of the tentacle and mouth of hydra.

Medindo alimentação resposta induzida por glutationa em Hydra

Hydra is among the most primitive organisms possessing a nervous system and chemosensation for detecting reduced glutathione (GSH) for capturing the prey. ...

Cardiac Pressure-Volume Loop Analysis Using Conductance Catheters in Mice

Cardiac pressure-volume loop analysis is the &#8220;gold-standard&#8221; in the assessment of load-dependent and load-independent measures of ventricular systolic and diastolic function. Measures of ventricular contractility and compliance are obtained through examination of cardiac response to changes in afterload and preload. These techniques were originally developed nearly three decades ago to measure cardiac function in large mammals and humans. The application of these analyses to small mammals, such as mice, has been accomplished through the optimization of microsurgical techniques and creation of conductance catheters. Conductance catheters allow for estimation of the blood pool by exploiting the relationship between electrical conductance and volume. When properly performed, these techniques allow for testing of cardiac function in genetic mutant mouse models or in drug treatment studies. The accuracy and precision of these studies are dependent on careful attention to the calibration of instruments, systematic conduct of hemodynamic measurements and data analyses. We will review the methods of conducting pressure-volume loop experiments using a conductance catheter in mice.

Cardiac pressure-volume loop analysis is the most comprehensive way to measure cardiac function in the intact heart. We describe a technique to perform and analyze cardiac pressure volume loops, using conductance catheters.

Cardiac Pressão-Volume Análise loop usando Conductance Cateteres em Ratos

Cardiac pressure-volume loop analysis is the &#8220;gold-standard&#8221; in the assessment of load-dependent and load-independent measures of ventricular ...

Measuring the Stiffness of Ex Vivo Mouse Aortas Using Atomic Force Microscopy

Arterial stiffening is a significant risk factor and biomarker for cardiovascular disease and a hallmark of aging. Atomic force microscopy (AFM) is a versatile analytical tool for characterizing viscoelastic mechanical properties for a variety of materials ranging from hard (plastic, glass, metal, etc.) surfaces to cells on any substrate. It has been widely used to measure the stiffness of cells, but less frequently used to measure the stiffness of aortas. In this paper, we will describe the procedures for using AFM in contact mode to measure the ex vivo elastic modulus of unloaded mouse arteries. We describe our procedure for isolation of mouse aortas, and then provide detailed information for the AFM analysis. This includes step-by-step instructions for alignment of the laser beam, calibration of the spring constant and deflection sensitivity of the AFM probe, and acquisition of force curves. We also provide a detailed protocol for data analysis of the force curves.

Measuring the Stiffness of Ex Vivo Mouse Aortas Using Atomic Force Microscopy

We present detailed protocols for isolation of aortas from mouse and measurement of their elastic modulus using atomic force microscopy.

Medindo a rigidez<em&gt; Ex Vivo</em&gt; Mouse aortas Usando Microscopia de Força Atômica

Arterial stiffening is a significant risk factor and biomarker for cardiovascular disease and a hallmark of aging. Atomic force microscopy (AFM) is a ...

The Mouse Isolated Perfused Kidney Technique

The mouse isolated perfused kidney (MIPK) is a technique for keeping a mouse kidney under ex vivo conditions perfused and functional for 1 hr. This is a prerequisite for studying the physiology of the isolated organ and for many innovative applications that may be possible in the future, including perfusion decellularization for kidney bioengineering or the administration of anti-rejection or genome-editing drugs in high doses to prime the kidney for transplantation. During the time of the perfusion, the kidney can be manipulated, renal function can be assessed, and various pharmaceuticals administered. After the procedure, the kidney can be transplanted or processed for molecular biology, biochemical analysis, or microscopy.
This paper describes the perfusate and the surgical technique needed for the ex vivo perfusion of mouse kidneys. Details of the perfusion apparatus are given and data are presented showing the viability of the kidney&#39;s preparation: renal blood flow, vascular resistance, and urine data as functional, transmission electron micrographs of different nephron segments as morphological readouts, and western blots of transport proteins of different nephron segments as molecular readout.

The mouse isolated perfused kidney (MIPK) is a technique for keeping a mouse kidney under ex vivo conditions perfused and functional for 1 hr. The buffers and surgical technique are described in detail.

A técnica de rim perfundidos rato isolado

The mouse isolated perfused kidney (MIPK) is a technique for keeping a mouse kidney under ex vivo conditions perfused and functional for 1 hr. This is a ...