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Traditional in vivo nucleic acid delivery systems, such as gene gun delivery, suffer from poor efficiency. The AgilePulse In Vivo System effectively introduces DNA or RNA into the target tissue, representing a powerful and safe means for stimulating an immune response that recognizes and eliminates target molecules in the body.

Upon implantation, mammalian embryos undergo major morphogenesis and key developmental processes such as body axis specification and gastrulation. However, limited accessibility obscures the study of these crucial processes. Here, we develop an ex vivo Matrigel-collagen-based culture to recapitulate mouse development from E4.5 to E6.0. Our system not only recapitulates embryonic growth, axis initiation, and overall 3D architecture in 49% of the cases, but its compatibility with light-sheet microscopy also enables the study of cellular dynamics through automatic cell segmentation. We find that, upon implantation, release of the increasing tension in the polar trophectoderm is necessary for its constriction and invagination. The resulting extra-embryonic ectoderm plays a key role in growth, morphogenesis, and patterning of the neighboring epiblast, which subsequently gives rise to all embryonic tissues. This 3D ex vivo system thus offers unprecedented access to peri-implantation development for in toto monitoring, measurement, and spatiotemporally controlled perturbation, revealing a mechano-chemical interplay between extra-embryonic and embryonic tissues.

Vivo Y27 System Update Download

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Investigators have also reported use of an intravenous gold-coated stainless steel medical wire with a hydrogel layer covalently coupled with antibodies against epithelial cellular adhesion molecule (EpCAM) protein (GILUPI CellCollector)20,21,22,23,24,25. However, physiologic variations between patients affecting blood flow and affinity make it difficult to standardize quantitative interpretation of CTCs by time of insertion. Similarly, a recent study has demonstrated in vivo capture of non-small cell lung cancer cells injected into a porcine model, using a flexible magnetic wire (MagWIRE)26. However, the approach requires pre-injection of EpCAM coated magnetic particles to label CTCs which limits its long-term application due to possible systemic exposure of iron overload.

To address the shortcomings of current approaches, we developed a temporary indwelling, intravascular aphaeretic CTC isolation system that can potentially be worn by a patient for several hours and through which a relatively large volume of blood can be interrogated. We demonstrate the feasibility of this device in ex vivo experiments, and we have performed proof-of-principle investigations of its potential clinical application for in vivo CTC capture in a canine model.

The in vivo aphaeretic CTC isolation system. a, b Schematic overview of the indwelling intravenous system by functional components (a) and manifold (b). c Application of a dual lumen catheter for in vivo CTC isolation

Despite the promising clinical implications of monitoring CTCs as a liquid biopsy, current CTC assays are hampered by the requirement that only a limited amount of blood can be collected at a single time-point. The indwelling intravascular aphaeretic CTC collection system we describe interrogates a larger blood volume in vivo over an extended period of time for capture and subsequent analysis of CTC. We liken this device to a Holter monitor commonly used by cardiologists to monitor cardiac arrhythmias over a prolonged period of time vs. at a single time-point provided by a rhythm strip.

In summary, we show that a temporary indwelling intravascular aphaeretic system can enable long-term operation in vivo to continuously harvest large quantities of CTCs, by re-transfusing the remaining blood products after the isolation procedure with minimal cell loss nor patient burden. Successful demonstration in canine models confirms the feasibility of our approach for future interventional clinical studies. The flexibility of the current system design can also be combined and adapted with various CTC enrichment methodologies or biochemical sensors that requires real-time analytical information from the blood. Finally, high numbers of CTCs obtained from large volume of blood screening will significantly reduce errors in determining the disease status and allow multiple characterization of CTCs to gain insight into their molecular and functional role realizing the full potential of a true liquid biopsy.

T.K., S.N., D.H.T., and D.F.H. conceived and designed the study. T.K. and C.R.O. built and tested the system with wireless controller. T.K., A.W. and K.J.S. fabricated the chips and performed the ex vivo CTC isolation experiments. T.K., Y.W. and D.H.T. performed the in vivo CTC capture experiments in canine models. D.H.T. provided canine blood samples and collected the patient data. L.C. and C.P. provided essential protocols and technical support. T.K., S.N., D.H.T. and D.F.H. analyzed the data and interpreted the results. T.K., S.N. and D.F.H. co-wrote the manuscript. All authors discussed and edited the manuscript.

Our in vivo recording systems connect to almost all commecially available probes. We recommend the use of probes from NeuroNexus or ATLAS Neuroengineering. Please see their company websites for probe specifications and order information.

Quasi Vivo is an advanced interconnected cell culture flow system, specifically designed to provide a solution to the major problems academic and industry researchers encounter when using conventional in vitro systems and in vivo research.

With over 20 years of experience Berthold Technologies is one of the pioneers of in vivo imaging. Our first system, the Luminograph, enabled scientists to detect low light emission in organisms. This opened up the opportunity of monitoring reporter genes in animals and plants in a non-invasive way. Today, our in vivo imaging systems offer the performance and features you need for your research, with optimized versions for analysing plants or laboratory animals.

The NightOWL II is the first imager with a motor-driven camera inside the cabinet. To cover the broadest range of in vivo imaging applications, the NightOWL II provides a wide variety of accessories, including heating tables, gas anaesthesia unit, animal isolation chambers, animal beds, and more

The NightSHADE evo LB 985N In Vivo Plant Imaging System is a modular, easy to use optical imaging system dedicated to in vivo analysis of plants. Equipped with a multi-position camera, it enables sensitive luminescence and fluorescence monitoring in tissues, seedlings and whole plants

In vivo imaging is the non-invasive visualization of living organisms for research or diagnostic purposes. Generally speaking, this method can be divided in two key areas: anatomical/morphological imaging and molecular imaging. In molecular imaging cellular function or molecular processes are visualized, normally using biomarkers. While in anatomical imaging no marker is used and visualization is based on the intrinsic properties of the tissues and organs being observed, such as the attenuation of X-rays in the case of computed tomography, molecular imaging very often uses biomarkers labeled with bioluminescence or fluorescence.

The IVIS Spectrum in vivo imaging system combines 2D optical and 3D optical tomography in one platform. The system uses leading optical technology for preclinical imaging research and development ideal for non-invasive longitudinal monitoring of disease progression, cell trafficking and gene expression patterns in living animals.

For advanced fluorescence pre-clinical imaging, the IVIS Spectrum has the capability to use either trans-illumination (from the bottom) or epi-illumination (from the top) to illuminate in vivo fluorescent sources. 3D diffuse fluorescence tomography can be performed to determine source localization and concentration using the combination of structured light and trans illumination fluorescent images. The instrument is equipped with 10 narrow band excitation filters (30nm bandwidth) and 18 narrow band emission filters (20nm bandwidth) that assist in significantly reducing autofluorescence by the spectral scanning of filters and the use of spectral unmixing algorithms. In addition, the spectral unmixing tools allow the researcher to separate signals from multiple fluorescent reporters within the same animal.

Accidental injury to the cardiac conduction system (CCS), a network of specialized cells embedded within the heart and indistinguishable from the surrounding heart muscle tissue, is a major complication in cardiac surgeries. Here, we addressed this unmet need by engineering targeted antibody-dye conjugates directed against the CCS, allowing for the visualization of the CCS in vivo following a single intravenous injection in mice. These optical imaging tools showed high sensitivity, specificity, and resolution, with no adverse effects on CCS function. Further, with the goal of creating a viable prototype for human use, we generated a fully human monoclonal Fab that similarly targets the CCS with high specificity. We demonstrate that, when conjugated to an alternative cargo, this Fab can also be used to modulate CCS biology in vivo, providing a proof of principle for targeted cardiac therapeutics. Finally, in performing differential gene expression analyses of the entire murine CCS at single-cell resolution, we uncovered and validated a suite of additional cell surface markers that can be used to molecularly target the distinct subcomponents of the CCS, each prone to distinct life-threatening arrhythmias. These findings lay the foundation for translational approaches targeting the CCS for visualization and therapy in cardiothoracic surgery, cardiac imaging, and arrhythmia management.

Labeling of the CCS using systemic mCntn2-800. To assess the feasibility of labeling the CCS in vivo, we conjugated IRDye800CW NHS ester, an NIR dye already in clinical use (10), to a commercially available polyclonal antibody directed against Cntn2, an extracellular marker previously shown to be expressed specifically within the CCS of mice and humans (Figure 1A) (9, 11). We injected wild-type adult mice intravenously with a single dose (75 g) of either mCntn2-800 or control IgG-800 (i.e., nonspecific IgG conjugated to IRDye800CW NHS ester), harvested the hearts and all other major organs after 72 hours, and imaged them using a closed-field NIR imaging system (Figure 1B). NIR signal was detected expectedly within the liver and kidneys, similar to prior reports of metabolism and clearance of other NIR imaging agents (12). Notably, mCntn2-800 signal was not detected within the brain tissue despite it being the only other major organ besides the CCS known to express Cntn2 (13), consistent with an intact blood-brain barrier (Figure 1C). 006ab0faaa