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LSM Mp4 [Extra Quality]

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LSM mp4


The jury, made up of British composer John Rutter, Latvian mezzo-soprano Elīna Garanča and Swiss conductor Nicolas Fink, praised the powerful performance that showed strength in timbre, tone, power and a nuanced delivery.

Photo: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Kārlis Miksons/LTVPhoto: Ekrānuzņēmums no LTV

The Elyra combines three dimensional (PRILM Technology), dual channel dSTORM/PALM super-resolution imaging with standard confocal microscopy to study protein localization and geometry at the nanoscale level. The setup is able to achieve an experimental resolution of 30 nm laterally, and 70 nm axially. The microscope is also suitable for live imaging of fluorescent-tagged transgenic cell lines.

Inverted super-resolution confocal microscope used for studying protein localization, motility and colocalization using traditional antibody labelling. Additionally, the microscope is fitted with an incubation chamber for live-cell imaging experiments requiring control of temperature and O2/CO2 levels. The LSM 800 enables simultaneous imaging of up to 4 different proteins. With Airyscan processing, the setup is able to achieve a resolution of 120 nm laterally and 350 nm axially, far exceeds that of a conventional confocal microscope.

Inverted confocal microscope used for studying Ca2+ dynamics in live cells. The 7 Live provides high speed scanning and gives acquisition rates of 1000 images of 512 x 512 pixels in less than 10 seconds. The microscope is set up with a perfusion system (37C), and field stimulation. Flash photolysis equipment is also available.

This fluorescence system enables wide-field fluorescence detection such as Fluo-4 (Easy Ratio Pro) by high-speed camera (Orca Flash 4.0, Hamamatsu Photonics) with WarpDrive interface controller. The setup is also equipped with a Cell Tester system (World Precision Instruments), enabling concomitant measurements of local cellular Ca2+ dynamics as well as contractile force development at varying sarcomere lengths. Additionally, the setup contains a heated perfusion system, as well as equipment for electrophysiology (Axon Axoclamp 200B) and field stimulation.

This inverted point-scanning confocal microscope has modest spatial and temporal resolution, and is commonly employed for both imaging of ionic fluorophores and static imaging protein localization / colocalization. This setup is also equipped with whole-cell fluorescence with PMT-based detection (Ratio Master, Photon Technology International), a heated perfusion system, electrophysiology (Axon Axoclamp 2B).

Zeiss Axio Observer / PTI Wide-Field FluorescenceThis inverted wide-field microscope is employed for studying ionic dynamics in live cells. Whole-cell ratiometric fluorescence (for example Fluo-4, Fura-2, SBFI) can be measured using the Ratio Master system from Photon Technology International employing Felix software. The setup is equipped with a heated perfusion system, equipment for measuring cell shortening (Crescent electronics) and electrophysiology (Axon Axoclamp 2A) and for exciting cells by field stimulation.

Huygens deconvolution software from Scientific Volume ImagingThe core facility is licensed for use of deconvolution software from SVI which improves image quality by reversing optical distortion in the image.

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CRITICAL: Commercially available pre-coated PDL coverslips rarely show lot number variation. When you have an issue with cell adhesion for coated coverslips, you should check and discuss coating quality with the supplier.

Note: When expressing the p3XFlag-SEP-DAGLα plasmid, the Flag tags were designed to localize to the intracellular region, and SEP was located on the extracellular surface. This structure makes it possible to confirm whether the protein expressed on the membrane is properly positioned through immunocytochemistry (Figure 5).

CRITICAL: The diameter of the pinhole affects resolution in X, Y and Z. The diameter of the first minimum of the airy disk is referred to as one airy unit. The size of the airy unit at the pinhole depends on the objective lens NA, the wavelength of the fluorescent light, and any magnification up to the pinhole.

CRITICAL: Exposing excessive laser-light damages cells and leads to death. To obtain the maximum bleaching effect with minimal stimulation, the membrane was stimulated for different periods, and the optimal effect was confirmed at 5 seconds (Figure 3B).

CRITICAL: Resolution depends on the diameter of the pinhole. The resolution in X and Y is determined by the distance from the center of the airy disk, and it depends on the wavelength of light (λ) and the NA. For a confocal system, the X and Y resolution equation is

Alternatives: As we mentioned above, PDL-pre-coated coverslips (GG-12-1.5-PDL; Neuvitro Corporation) could be used if the quality is tested. To perform the FRAP, similar or advanced models of Nikon C2+ confocal microscope system such as Nikon A1R, LSM 980 (ZEISS), and STELLARIS 8 (Leica) can be used alternatively. However, it is recommended to test FRAP conditions when using other systems.

CRITICAL: Trypsin treatment for 10 min digests various extracellular matrix proteins allowing easy mechanical dissociation. However, neurons are weakened and can die quickly. To increase the survival rate of neurons, resuspend cells by pipetting up and down with a 1 mL tip placed close to the bottom of the conical tube. Pipetting should be done gently and carefully, never performing more than 10 repetitions.

CRITICAL: The 12-well plate and medium used for maintenance of the neuronal cells put back into the incubator. After transfection, the coverslips will be returned to the original 12-well plate.

CRITICAL: To ensure experimental reproducibility, only healthy pyramidal neurons are considered, and subjects with abnormal morphology or weak fluorescence intensity are excluded (Figure 6). Troubleshooting, problem 4.

(D) FRAP curve of SEP-DAGLα in primary cultured neurons. Normalize the intensity of FITC by TRITC. One-phase exponential equations fitted the curves. t1/2: the half time of equilibrium; IF: immobile fraction; MF: mobile fraction.

XFile3 simplifies your file transfer and archive needs during a live production. With higher levels of efficiency and automation, you can archive, transform and restore your selected content - in any format and from multiple destinations. This software application provides complete control and visibility of all file transfers from one single intuitive interface, whether the user is archiving locally or through connected live workflows enabled by C-Next.

XFile3 allows immediate access and archiving of EVS server content during the event. You can also live stream your content directly to your post-production platforms for real-time collaboration from one central interface.

"At EVS we know that XFile has been a great tool in live production environments for years now, as there is always someone walking into the truck that wants to play back clips just seconds later" Brady Jones - Lead EVS LSM-VIA ambassador

In the last years we have seen the increased need for more tools in XFile. Replay operators or media managers need to be able to not only import last minute content but also to import content coming from other shows. To facilitate this, EVS has been increasing the codec compatibility. This way, EVS solutions can be easily integrated with each other as well as with other 3rd party systems and post-production tools.

For Multicamera recording, XFile3 allows real-time streaming of all camera angles from the server to your storage using an intuitive user interface and including crash-record capabilities and time-based scheduling of these streams.

Besides sharing content via hard drives and USB sticks, XFile3 also gives you the possibility to share content through cloud storage. For example, you can push content to AWS S3 storage, so another show can download the content from their XFile3 interface.

Beside the regular connections and drives that can be attached to the XFile hardware, EVS also validated Seagate LYVE storage which can be rented on demand. This high performance cloud storage service focuses on secure transport of your content. For benchmarks or more information on the offering, reach out to our sales team. 041b061a72


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