Sep 15, 2011 on bd pathway 855 bioimager system u sing bd attovision tm softw are. Thr172 cell signaling t echnology was used in this set of images. Pdf raptor, a positive regulatory subunit of mtor complex 1. Bd attovision software was used to automatically identify and quantify nuclei stained with hoechst 33342 invitrogen, h3570, gfpp65, and mcherry fluorescence. The 855 bioimager has an integrated environmental control chamber that allows for imaging. This study proves that downregulation of signaling pathway of fak is involved in ethanolinduced apoptosis in human skin fibroblasts.
Preparation and storage store undiluted at 4c and protected from prolonged exposure to light. Bdpathway 855 the bd pathway systems from bd biosciences offer an integrated solution encompassing instrumentation, software and reagents. Bd pathway systems provide fluorescence intensity measur ements and temporal and morphological imaging, including subcellular imaging to detect cellular events. Image analysis in highcontent screening semantic scholar. Image quality, segmentation, object features, software. A comparison of fluorescent images generated by the three staining methods. Here, we explore how pathogenic mutations and the inhibition of lrrk2 kinase activity affect cytoskeleton dynamics in mouse and human cell systems. Bd pathway 855 quick user guide to the automatic fluorescent microscopehighthroughput screener hts lasse evensen march 2012. Figure 8 the bd biosciences pathway 855 highcontent bioimager for. Preparation and storage the monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. For imaging, islet cells were immersed in hepesbuffered solution mm.
The bd pathway 435 highcontent bioimager is a compact bench top imager compatible with multiwell plates and slides. The bioimager was equipped with a 48810nm enhanced gfp egfp excitation filter, a 380 10nm dapi excitation filter, a 515nm longpass lp egfp. The bd pathway software provides a suite of integrated tools to accomplish the tasks of. Acquired images were processed using attovision software bd biosciences. The bd pathway 855 bioimager is a high content cell analyzer designed for superior performance and flexibility. Connexin 43 and metabolic effect of fatty acids in stressed. Huvecs, skin fibroblasts seed cells in 96well clear bottom tissue culture plates for imaging applications becton dickinson. Cb 2 agonist capable to prevent macrophage activation in vivo. The percentage of dead cells in the presence of inhibitors was determined by using the bd attovision v1.
Highthroughput imaging method for direct assessment of gm1. The liquid handling and detection module are highly integrated by a mitsubishi rv2aj. Visualization of fixedpermeabilized cultured cells by bd pathway bioimager 855. The pipeline pilot imaging software bundle can also help to process the data generated from this. Bd pathway highcontent analyzers technical specifications. Becton dickinsons bd pathway 855 highcontent bioimager is one of the.
Jordy stichelmans phdstudent vrije universiteit brussel. Nerve growth factor induces neurite outgrowth of pc12. This system can perform a wide range of fluorescencebased kinetic and endpoint biological. Bd pathway tm 855 highcontent bioimager labmakelaar. It can handle specimen formats from single slides to multiwell plates. The detection modules include the bd pathway 855 bioimager, epic system, tecan safire 2 reader, aquest currently analyst gt plate reader, icr8000 atomic absorption spectrometer, spectramax 340 reader, and las3000 fuji imaging station. Pdf the active form of the metabolic sensor ampactivated.
Silibinin suppresses emtdriven erlotinib resistance by. Estimation of the cx43 protein amount in huvecs and isolated mitochondria. The gfpp65 and tnf promotermcherry reporters in raw g9 cells were imaged with a bd pathway 855 bioimager bd biosciences. The laserbased autofocus capability of the bd pathway systems enables rapid acquisition of highquality images. Bd pathway 855 bioimager hardware upgrades subject. The bd pathway 855 bioimager is a selfcontained system used to obtain superior image quality in highcontent live or fixed cell analysis. In order to test the sensitivity and the capacity to detect eyfp and ecfp separately using bd pathway 855 bioimager, the cocultured cells were photographed for the two. Pathway 855 bioimager bd biosciences imaging platform is equipped. Becton dickinsons bd pathway 855 highcontent bioimager is one of the most capable imaging solutions available for live or fixed cell samples. The system enables a broad array of fluorescencebased biological applications such as neurite outgrowth analysis, angiogenesis, protein translocation.
The monoclonal antibody was purified from tissue culture supernatant or ascites by affinity chromatography. Fixedpermeabilized cultured cells bd pathway bioimager 855. It can automatically image multiple sites per slide or well and every well of a multiwall plate. Differential nuclear staining assay for highthroughput. Highthroughput imaging method for direct assessment of. Dell precision 390 workstation with windows xp operating system. Nerve growth factor induces neurite outgrowth of pc12 cells. Imaging studies were performed in 96well plates bd falcon using the bd pathway 855 bioimager microscope bd biosciences.
Bd pathway t m 855 bioimager system and merged using bd attovisiontm software. Attovision software flexible software for both kinetic and endpoint experimentation provides a broad range of acquisition and analysis tools. Development of a cellbased fluorescence polarization. It combines superior image quality, flexible image capture, and livecell analysis to address a wide range of. Iiitubulin immunofluorescence b and hcs cellmask red c fluorescence images of cell bodies and neurites were acquired on the bd pathway 855 bioimager.
It will rapidly record highresolution fluorescence images from multiwell plates. Bd pathway 435 confocal bioimager from bd biosciences. Igf1repithelialtomesenchymal transition emt crosstalk. Bd pathway 435 bioimager biomedical microscopy core bmc uga. All liquid handling for treatment and staining was carried out by a robotic workstation and the bd pathway 855 cell imaging platform was used for automated image acquisition. A rapid, inexpensive high throughput screen method for. The company has two bioimaging systems currently on the market, the bd pathway 435 and 855 bioimagers. B was added prior to washing and staining with bd bioimaging.
For corning huvec2, hmvec, and hmec1, 70 to 80% confluence is recommended. Images show representative metforminnaive a431 cells and metforminrefractory a431 metr cells captured in different channels for phosphoigf1r tyr1161 red and hoechst 33,258 blue with a 20. Bd pathway bioimaging systems brochure bd biosciences. Connexin 43 and metabolic effect of fatty acids in. Binocular eyepieces the bd pathway bioimager is the only system in its class with. Protective influence of hyaluronic acid on focal adhesion. Dispersed islet cells were plated at a density of 20,000 cells per well in blacksided 96well plates coated with polydlysin, and cultured overnight in cmrl medium. Pathway qbiomarker somatic mutation pcr arrays, which com bine allelespecific amplification and 5 9 hydrolysis probe detection to detect as little as 0. Both the benchtop bd pathway 435 and the standalone bd pathway 855 provide high performance and ease of use to improve workflow and productivity, al ong with advanced automation features. The highmobility group iy transcription factor is involved. Jan 01, 2009 the company has two bioimaging systems currently on the market, the bd pathway 435 and 855 bioimagers. A dualcolor fluorescencebased platform to identify.
We generated and characterized a novel transgenic mouse model expressing. Light from a mercury metal halide lamp introduced through a liquid light guide provides illumination from 360 nm to 700 nm. In vivo cell monitoring bd pathway bioimager 855 jumc. Sep 24, 2011 connexin 43 protein was visualized by using a 1.
The bd pathway was designed to provide highresolution, automated confocal imaging with sophisticated imaging software to assist in developing better cellbased assays. The bd pathway 855 bioimager system and its associated attovision v1. The antimalarial chloroquine overcomes primary resistance. All the imaging data were analyzed with attovision software package. The powerful acquisition software included with the microscope allows users to.
Unaltered, exemplar tiff files were retrieved with imagej software and the yellow. Four objectives 4x, 10x, 20x and 40x are available to capture images in an ample variety of montages contiguous frames and confocal z stack acquisition mode. The 855 bioimager has an integrated environmental control chamber that. The bd pathway systems from bd biosciences offer an integrated solution encompassing instrumentation, software and reagents. Analysis can be conducted in multiple modes, including kinetic imaging, fluorescence intensity measurements, read more. Aid 1224826 genomewide sirna screen of genes regulating. A new optical upgrade package is now available to optimize the system s ability to capture fluorescence images.
Images were acquired on the bd pathway 855 highcontent bioimager bd biosciences. The pipeline pilot imaging software bundle can also help to process the data generated from this system. This antibody also stains a549 atcc ccl185 and hela atcc ccl2 cells, and it works with either cold methanol or triton x100 permeabilization see recommended assay procedure. Some other commonly used imaging software packages for measuring the extent of tube formation include bd pathway 855 bioimager with sophisticated image and data analysis algorithms. Autophagy facilitates the development of breast cancer. Bd pathway systems the benchtop bd pathway 435 and standalone bd pathway 855 feature precision optics with flexible and easytouse controls and comprehensive acquisition and analysis software.
The bd pathway 855 system offers the ultimate in flexibility for highcontent imaging of live and fixed. The rect angular regions white lines are enlarged an d shown as high magni ca tion insets in the rig ht parts. Nov 18, 2015 the cells were then washed once with pbs and imaged on a bd pathway 855 bioimager using a 20. A trueoptical spinning disk allows the operator to. The intuitive and customizable software provides for both readytorun and fully customizable applications. The intracellular mobility of these fluorescent tracers can be investigated using fluorescence recovery after.
Subsequently, images were segmented and analyzed for gfp granularity or hmgb1gfp nuclear intensity using the bd attovision software version 1. Mutations within the lrrk2 gene have been identified in parkinsons disease pd patients and have been implicated in the dysfunction of several cellular pathways. The confocal mode enables a bd pathway instrument to deliver. Becton dickinson s bd pathway 855 highcontent bioimager is one of the most capable imaging solutions available for live or fixed cell samples. The analysis of fluorescence imaging using bd pathway 855 highcontent bioimager showed the inhibition of fak migration to the cell nucleus, depending on the increasing concentration of ethanol.
After dye addition and incubation, images were captured with two software predetermined excitationemission filter sets. Visualization and in vivo monitoring of live cells by bd pathway bioimager 855. Bd pathway 435 confocal bioimager from bd biosciences bioimaging,the bd pathway 435 is an affordable, compact benchtop imager compatible with multiwell plates and slides, providing a broad array of automated imaging capabilities. The system rapidly acquires highresolution images with highperformance laser autofocus, true confocal optics, flexible computercont,biological,biology supply. Other aminereactive dyes many of the ph indicators described in probes useful at nearneutral phsection 20. This sytem offers highcontent imaging of fixed cells. Bd pathway bioimager from bd biosciences selectscience. The bd pathway 855 system offers the ultimate in flexibility for highcontent imaging of live and fixed cells. Phasecontrast left and immunofluorescence right images were obtained and merged on bd pathway 855 bioimager system according the recommended assay procedure bd attovision software.
It can collect transmitted and fluorescence light images. Highcontent imager, bd pathway 855 molecular imaging. Images were merged on a bd pathway 855 bioimager system with bd attovision software. The bd pathway bioimager is an automated, confocal, realtime, single cell kinetic and endpoint imaging system that has been integrated into a single, compact unit. Our main products include custom fluorescence, biological, metallurgical and polarizing microscopes, microscopy cameras and digital scanners such as pathology slide scanner, gel scanner and 3d imager. Bd pathway 855 quick user guide to the automatic fluorescent microscopehighthroughput screener hts lasse evensen march 2012 2 starting up the instrument and imaging software insert the desired objective turn on the main switch turn on the computer located in the cabinet. Tracing fluid transport with dextrans fluorescent dextrans are important tools for studying the hydrodynamic properties of the cytoplasmic matrix.
The bd pathway 855 high content bioimager bd biosciences, san jose, ca was used for automated image acquisition. Free actin impairs macrophage bacterial defenses via. If mammalian target of metformin indirectly is mammalian. Figure shows representative microphotographs of wound healing assays 0. This standalone system offers exceptional image quality, adjustable image capture, and live cell analysis, enabling it to meet the needs of a variety of applications. Images show representative portions of skbr3 and tzbr cell cultures captured in different channels for lc3 red and hoechst 33258 blue with a 20. The bd pathway 855 2006model is a fully automated microscope that provides the option of automated image acquisition and data analysis on multiwell plates 96 and 384. Extracts from cells transfected with the empty vector pcmv6 lane 1. Flexible and powerful, the bd pathway systems enable the researcher to choose between the ease of readytorun applications to developingyourown customized assays.
Bd pathway bioimager system bd biosciences this multiwell 96 and 384 plate reader confocal microscope system has stationary stage with movable optics and it is assisted with attovison software to capture images and data analysis. It combines superior image quality, flexible image capture, and livecell analysis to address a wide range of applications. Of note, tzb exposure further increases autophagosome lc3 pattern. Bd pathway 435 bioimager biomedical microscopy core bmc. Extracts from cells transfected with the empty vector pcmv6 lane 1, nontransfected cell ntc. Images were analyzed using cellprofiler software 18 and mcherry features were tabulated from cells, including granularity, texture, and average mcherry intensity.
Aug 22, 20 the antimalarial chloroquine overcomes primary resistance and restores sensitivity to trastuzumab in her2positive breast cancer. Purchased september 2008, the 855 is a versatile system for highcontent confocal imaging of live or fixed samples. Images show representative portions n 3 of untreated or tgf. With updated image analysis software featuring new tools and wizardstyle applications, the bd pathway 435 confocal bioimager simplifies the process of highcontent cellular analysis. Fixedpermeabilized cultured cells bd pathway bioimager.
657 1615 1200 1578 1305 475 1430 1526 849 793 99 1192 655 923 372 33 686 650 1431 1623 1515 772 259 1131 344 968 94 1289 311 929 458 1231 567