Metamorph microscopy automation and image analysis software. These image montages were then used for subsequent neurite outgrowth analysis. Immunostaining was performed 48 hours later, and neurite outgrowth was analyzed using metamorph neurite outgrowth software v7. The software will display the optical densities of a brightfield source image in a. Metamorph program is sending data to the spreadsheet because the metamorph program will be unable to send future information to the spreadsheet or to the correct position on the spreadsheet. Openmeta helps you bring models and analyses together to. Raw images were acquired using the minimax cytometer and stitched together using metamorph software. Our selection of metamorph as opposed to one of the many other image processing tools available was motivated simply by the fact that we already had access to the software. Analysis of neurite length was done by manually tracing the neurites using metamorph software universal imaging corporation, west chester, pa, u. Two small gtpases, rab and arf, are wellknown molecular switches that function in diverse membranetrafficking routes in a coordinated manner.
Stages of neuronal morphological development in vitroan. Optional application modules perform assayspecific analysis, such as counting nuclei or assessing cell cycle phases, in a simple wizardlike interface. Metamorph will open most tif files, however most analysis needs 16bit tifs. For application modules that are metamorph dropin applications, such as. The design of complex systems requires many engineering specialties and software tools. Cell migration is required for axonal guidance and neurite outgrowth and involves a series of highly coordinated and overlapping signalling pathways. To open a zstack generated in metamorph or another program or create a stack from a series of individual images. Gfp and pcdna3gfp control p software described in this document, including information contained in any. Neurite lengths were determined as the distance from the. Our team has a combined 40 years experience in building and integrating modeldriven engineering tools. The metamorph tools are designed to make the creation of electronic circuits as accessible as the creation of software apps. Oct 19, 2016 unless specified otherwise, neurite outgrowth was evaluated for a total of 80100 neurons per experimental point using the metamorph neurite outgrowth module, by taking 34 images containing 8.
Rab small gtpases are key regulators in membrane trafficking, and although several rab isoforms, e. Based on this input the software performed a cell segmentation step segmented tuj1 image and. The nonreceptor tyrosine kinase, focal adhesion kinase fak has. For the neurite outgrowth analysis, the semiautomated metamorph software molecular devices was applied to analyze average and longest process length. Developed in conjunction with researchers, metamorph offers a range of tools for a variety of applications such as. Metamorph softwares builtin stitching feature was used to create the image montages from the original data generated in softmax pro software figure 3. Quantitative neurite outgrowth measurement based on image. With the metamorph offline package, users can measure, analyze, and display multidimensional data acquired from a confocal system. Neurite outgrowth is mediated by the exocytosis of intracellular vesicles at the tips of elongating neuronal processes. Leica mm af provides a platform for neurite outgrowth experiments including both image acquisition and image analysis.
This biological phenomena is referred to as neurite outgrowth and is regulated by complex. Tbc1d12 is a novel rab11binding protein that modulates. Some modules handle very specific and challenging assays e. Each identified neuron has a number associated with it. We recommend capturing and saving images as 16 bit tifs. Oct 24, 2008 to understand the process of neurite outgrowth in the context of neuroregeneration, we used mouse neuroblastoma n1e115 as our model neuronal cell. Manual analysis of neuronal morphology is time consuming and becomes impractical for large datasets. Synaptotagmin 17 controls neurite outgrowth and synaptic. Optimising parameters for the differentiation of shsy5y. The software will display the optical densities of a brightfield source image in a scaled 8bit or 16bit image. Metamorph has the functionality of measuring the length of. Neurite outgrowth application module is designed to measure and characterize outgrowths, the extension of processes from the cell body, which are natural parts of neuronal development. Neuritequant does not require any additional commercial software products. Pituitary adenylate cyclaseactivating polypeptide prevents the effects of ceramides on migration, neurite outgrowth, and cytoskeleton remodeling anthony falluelmorel, david vaudry, nicolas aubert, ludovic galas, magalie benard, magali basille, marc fontaine, alain fournier, hubert vaudry, and bruno j.
For the analysis of neurite outgrowth in div3 hippocampal slice cultures various parameters were analysed using metamorph software using the track points function. Links to image analysis software duke light microscopy. To further tailor analysis to your researchoriented goals, journals can be amended to application modules, allowing customizing on any analysis. Neurite outgrowth apps menu 716 angiogenesis tube formation apps menu 726. Neurons create connections via extensions of their cellular body, which is referred to as neurite outgrowth and is regulated by complex intracellular signaling events. Neurite outgrowth in culture provides an easy way to determine the effects of a particular substrate or exogenous factor on neuron behavior. Metamorph for olympus is an advanced digital imaging software for capture, display, and analysis of biological images. Data that has been sent to a text file can be displayed using the view current type log function on the log menu. Through highly regulated mechanisms, young neurons undergo axonaldendritic polarization, and subsequent outgrowth of these neurites is essential to the establishment of synaptic connections that lead to brain function da silva and dotti, 2002.
Using metamorph softwares neurite outgrowth module, cell bodies and their outgrowths were identified, and measurements and analysis were reported. The synaptotagmin syt proteins have been widely studied for their role in regulating fusion of intracellular vesicles with the plasma membrane. Metamorph softwares neurite outgrowth image analysis module was used to. Neurite outgrowth is the extension of axonal processes from the cell body and is a natural part of early development. Briefly, a neurite tuj1 and nuclei pi image were loaded into the software. An open source toolkit for high content screens of. Understanding the signaling mechanisms driving neurite outgrowth provides valuable. Neurite outgrowth is an essential parameter for assessing not only a unique characteristic of neurons but also as. Whether youre adopting modeldriven engineering tools for your organization or looking to get more value from the models and tools that you already have, metamorph s team of experienced software developers can help. Metamorph software automatically calculates a variety of neurite outgrowth measurements. The module helps standardize results compared to traditional methods. Substrate topography determines neuronal polarization and. Pituitary adenylate cyclaseactivating polypeptide prevents. The neurite outgrowth application module is designed to facilitate the analysis of neurite outgrowth experiments.
Computer vision profiling of neurite outgrowth dynamics. Als2alsin regulates racpak signaling and neurite outgrowth. Understanding the signaling mechanisms driving neurite outgrowth. Sep 11, 20 cell migration is a fundamental biological process and has an important role in the developing brain by regulating a highly specific pattern of connections between nerve cells. We and others have previously shown that expression of the constitutive active mutant of rab35 rab35q67l in pc12 cells enhances ngfinduced neurite outgrowth, whereas its constitutive negative mutant rab35s22n inhibits it chevallier et al. Measuring neurite outgrowth with the spectramax minimax cytometer and metamorph software neurons create connections via extensions of their cellular body called dendrites or processes. Unless specified otherwise, neurite outgrowth was evaluated for a total of 80100 neurons per experimental point using the metamorph neurite outgrowth module, by. Cell migration is a fundamental biological process and has an important role in the developing brain by regulating a highly specific pattern of connections between nerve cells. Its component and circuit library, platform templates, and analysis tools help electronics designers build prototypes more quickly. Recycling endosomes are generally thought to play a central role in endocytic recycling, but recent evidence has indicated that they also participate in other cellular events, including cytokinesis, autophagy, and neurite outgrowth. Links to image analysis software duke light microscopy core. Whether youre adopting modeldriven engineering tools for your organization or looking to get more value from the models and tools that you already have, metamorphs team of experienced software developers can help. Metamorph is a great addition to a core facilitys confocal station.
Munc18 and munc regulate early neurite outgrowth omicx. Neurite article about neurite by the free dictionary. The lysosomal vesicleassociated soluble n ethylmaleimidesensitive factor attachment protein receptor tetanus neurotoxin insensitive vesicleassociated membrane protein tivampvamp7 was previously implicated in membrane fusion events mediating neurite outgrowth. Measuring neurite outgrowth with the spectramax minimax cytometer and metamorph software neurons create connections via extensions of their cellular body, which is referred to as neurite outgrowth and is regulated by complex intracellular signaling events. The metamorph tools are built on top of our openmeta framework. Module are seamlessly integrated with the flexibility of metamorph software and its advanced. With the flexibility of metaxpress software, researchers are able to choose. Please note, not all hardware drivers are compatible with 64bit systems. Neurite outgrowth, neurotoxicity molecular devices. Metamorph offers analysis tools to handle everything from simple intensity logging to advanced morphometry analysis, colocalization, fret, 3d measurements, and more.
In our experiments, changes in the ratio between total neurite length and neuronal cell body area usually resulted from altered neurite outgrowth. Neurite lengths were determined as the distance from the edge of the cell body to the growth cone tip. Inhibition or stimulation of neurite outgrowth is implicated in a broad range of cns disorders or injuries including stroke, parkinsons disease, alzheimers disease, and spinal cord injuries. Synaptic adhesionlike molecules salms promote neurite. Metamorph is commercial software and developed by mds for cellular image analysis. Metaxpress software and the neurite outgrowth application. Hamburg or custom journals implemented in the microscopy control software metamorph molecular devices, inc. Neurite outgrowth, this area displays the same description as the meta. Interestingly, taxol application to primary hippocampal neurons resulted in an increase in total neurite length and an apparent decrease in neuronal cell body area see 7.
Inhibition or stimulation of neurite outgrowth is implicated in a broad range of. A number of different application modules can be used to analyze data from. Topographical guidance of pscderived cortical neurons. It is available as part of the microscopes image acquisition system or as a standalone version. Neurite outgrowth is a fundamental event in the development and maintenance of synaptic connections in the nervous system. Built on the marketproven metamorph platform, this powerful package offers functionality, flexibility, and stability that meet the most demanding image acquisition and analysis needs. The nonreceptor tyrosine kinase, focal adhesion kinase fak has an. A role for synaptotagmin viiregulated exocytosis of. The study of neuronal morphology and neurite outgrowth has been enhanced by the. Several software packages based on the automated neuronal morphology and neurite outgrowth analyses have been published, such as neuritequant 8, neurite image quantitator neuriteiq 9.
Image analysis was performed using the metamorph neurite outgrowth plugin universal imaging. Metamorph premier 4 offline licenses ffsc 64bit, lsrc workstation 2, nan duke workstation and rp2 workstation tracking, 4dviewer, count nuclei, neurite outgrowth etc applications. Apr 26, 2006 neurite outgrowth is mediated by the exocytosis of intracellular vesicles at the tips of elongating neuronal processes. Images for neurite outgrowth experiments were analyzed using metamorph image analysis software. Open or build a stack to open a zstack generated in metamorph or another program or create a stack from a series of individual images. The metamorph microscopy automation and image analysis software automates acquisition, device control, and image analysis. Measuring neurite outgrowth with the spectramax minimax. Neurite outgrowth application module molecular devices. Images from each section of the well were loaded into a stack and stitched together into a montage using the softwares image processing features. While specialized commercial software applications are available to measure neurite outgrowth, such tools are usually not openly available for user customization beyond the supplied standard interface. Within metamorph, we used the neurite outgrowth and cell scoring modules to quantify the morphology of immunostained neurons from fluorescent images. Although rab35 and arf6 were independently reported to regulate the same cellular events, including endocytic recycling, phagocytosis, cytokinesis and neurite outgrowth, the.
Using a nuclear stain is beneficial to identify cell bodies in some cell types. Measuring neurite outgrowth with the spectramax minimax cytometer and metamorph software. Measurement of neurite outgrowth in high density cultures of. Metamorph imageanalysis software was used to quantify the length of the longest neurite. The software offers many userfriendly application modules for biologyspecific analysis. The lysosomal vesicleassociated soluble n ethylmaleimidesensitive factor attachment protein receptor tetanus neurotoxin insensitive vesicleassociated membrane protein tivampvamp7 was previously implicated in membrane fusion events mediating neurite outgrowth, but the. Quantitate neuron potential molecular devices crisel instruments. These actions of c2ceramide on cell motility and neurite outgrowth were.
It easily integrates dissimilar fluorescent microscope hardware and peripherals into a single custom workstation. If not convert them in image j or using the scale command 1. Research park ii image analysis workstation duke light. Metamorph from molecular devices now part of mds analytical. Neurite outgrowth and expression of progenitor and differentiation markers were analysed using the semiautomated software metamorph molecular devices, usa. Using metamorph softwares neurite outgrowth module, cell bodies and their outgrowths were identified, and measurements and analysis were reported automatically figure 4.
Developed in conjunction with researchers, metamorph offers a range of tools for. Openmeta helps you bring models and analyses together to get a better view of system performance and tradeoffs. The parameters of cellprofiler were suggested by the software developers and a service engineer from mds fine tuned the parameters for metamorph. Built on the marketproven metamorph platform, this powerful package offers functionality, flexibility, and stability that meet the. Neurite outgrowth angiogenesis tube formation time lapse zseries, and more an integrated imaging system for maximized control device automation for easy acquisition metamorph provides highend control for devices including microscopes, filter wheels, shutters, cooled ccd cameras, video cameras, monochromators, focus motors and piezo. Metamorph software runs under 32bit and 64bit operating systems. Metamorph gives users complete control over the type of data logged, as well as row and column label locations. Here, a specific neuron is selected yellow overlay and its associated measurements are highlighted blue in the table. The neurite outgrowth application module for metaxpress software is designed for the analysis of neurite outgrowth assays. Apr 25, 2008 metamorph from molecular devices now part of mds analytical technologies the good huge list of functions available. Measurements include total neurite outgrowth, total branches and cell bodies, straightness and more.
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