Tg(fli1:EGFP)y1 zebrafish were used in this study [34 (link)]. To block pigmentation, embryos were raised from 22 hpf in the presence of 0.003% N-Phenylthiourea (PTU; Sigma-Aldrich #P7629). 29 hpf embryos were placed in each well of a 24-well plate in 1 mL egg water containing 0.1% DMSO (control) or 500 µM TNP-470. Embryos were kept at 28.5 °C until imaging took place. For imaging, live embryos were anaesthetized using Tricaine mesylate and were mounted in 0.5% low-melting-point agarose (SeaPlaque, Lonza). Images were acquired using a Zeiss LSM700 confocal microscope and an Axio Imager M2 compound microscope equipped with a 40 × 1.0 NA water objective. Images were exported as TIFF files using ZEN 2009 LE software (Zeiss), and figures were assembled using the Adobe Photoshop CS4 software, n = 17.
Zen 2009 le software
Manufactured by Zeiss
Sourced in Germany
ZEN 2009 LE software is a imaging and microscopy platform developed by Zeiss. It provides basic image acquisition, processing, and analysis capabilities for microscopy applications.
Automatically generated - may contain errors
Lab products found in correlation
Seaplaque, by Lonza (3 mentions)
Lsm 700 confocal microscope, by Zeiss (1 mentions)
Axio imager m2 compound microscope, by Zeiss (1 mentions)
Axiocam mrc digital camera, by Zeiss (1 mentions)
Discovery v8 stereoscope, by Zeiss (1 mentions)
Axiovision 4, by Zeiss (1 mentions)
Lsm710 meta microscope, by Zeiss (1 mentions)
Lsm image browser software, by Zeiss (1 mentions)
Axio imager m2, by Zeiss (1 mentions)
Lsm 700, by Zeiss (1 mentions)
Axiocam mrc, by Zeiss (1 mentions)
4 protocols using zen 2009 le software
1
Zebrafish Angiogenesis Imaging Protocol
2
Photoconverting Embryos for Fluorescence Imaging
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To block pigmentation when imaging embryos older than 24 hpf, embryos were raised from 22 hpf in the presence of 0.003% N-Phenylthiourea (PTU; Sigma-Aldrich #P7629). Live embryos were anaesthetized using tricaine and mounted in 0.5% low-melting-point agarose (SeaPlaque, Lonza). For photoconversion, images of eye and OS region were acquired by minimal exposure using a 488 nm laser. A selected region of interest (ROI) was exposed to 405 nm laser scanning until all green fluorescence was converted to red, as determined by imaging with 488 nm and 555 nm lasers. The embryos were released from agarose and incubated at 28.5 °C in the dark until imaged again.
Images were acquired using Zeiss LSM700 confocal microscope and Axio Imager.M2 compound microscope or with Discovery.V8 stereoscope and AxioCam MRc digital camera (Zeiss). Microscope objective used were 40 × 1.0 NA water objective or 25 × 0.8 NA or 10 × 0.3 NA. Images were exported as JPEG or TIFF files using ZEN 2009 LE software (Zeiss) and figures were assembled using Adobe Photoshop CS4 software.
Images were acquired using Zeiss LSM700 confocal microscope and Axio Imager.M2 compound microscope or with Discovery.V8 stereoscope and AxioCam MRc digital camera (Zeiss). Microscope objective used were 40 × 1.0 NA water objective or 25 × 0.8 NA or 10 × 0.3 NA. Images were exported as JPEG or TIFF files using ZEN 2009 LE software (Zeiss) and figures were assembled using Adobe Photoshop CS4 software.
3
Quantitative Imaging of Adipocyte Clusters
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For laser-scanning microscopy studies, freshly isolated adipocyte clusters, were used immediately after 1-hour incubation at 37°C with 22 μmol/L QD675 in NaCl 0.9%. The high concentration was chosen to ensure a complete saturation in the labeling process and does not represent a suitable dosage for in vivo experiments. As a control group, adipocyte clusters without QDs incubation were used. Labeled and unlabeled adipocyte clusters were placed into Lab-Tek 8 chambered cover glasses (Nalge Nunc, Naperville, Ill). Confocal laser-scanning microscopy was carried out with a LSM 710 META microscope (Carl Zeiss, Jena, Germany) equipped with gas lasers emitting lines at 364 nm (argon), 488 nm (argon), and 633 nm (helium/neon laser). Pictures were taken with a plan-Apochromat 63Â/1.40 Oil differential interference contrast (DIC) objective.
For DNA and consecutive cell nucleus marking, DAPI was used. Fluorescence detection of QD675 CdTe QDs was achieved with a 650-nm longpass filter and laser excitation at 633 nm. 4′,6-Diamidin-2phenylindol was spotted with a 385-to 470-nm bandpass filter and excitation at 364 nm. Data were acquired and analyzed with AxioVision 4.6 software, LSM image browser software, and ZEN 2009 LE Software (Carl Zeiss, Jena, Germany).
For DNA and consecutive cell nucleus marking, DAPI was used. Fluorescence detection of QD675 CdTe QDs was achieved with a 650-nm longpass filter and laser excitation at 633 nm. 4′,6-Diamidin-2phenylindol was spotted with a 385-to 470-nm bandpass filter and excitation at 364 nm. Data were acquired and analyzed with AxioVision 4.6 software, LSM image browser software, and ZEN 2009 LE Software (Carl Zeiss, Jena, Germany).
4
Embryo Imaging with PTU Blocking
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To block pigmentation when imaging embryos older than 24 hpf, embryos were raised from 22 hpf in the presence of 0.003% N-Phenylthiourea (PTU; Sigma-Aldrich #P7629). Live embryos were anaesthetized using tricaine and mounted in 0.5% low-melting-point agarose (SeaPlaque, Lonza).
Images were acquired using Zeiss LSM700 confocal microscope and Axio Imager.M2 compound microscope or with Discovery.V8 stereoscope and AxioCam MRc digital camera (Zeiss). Microscope objectives used were 40x 1.0 NA water objective or 25x 0.8 NA or 10x 0.3 NA. Images were exported as JPEG or TIFF files using ZEN 2009 LE software (Zeiss) and figures were assembled using Adobe Photoshop CS4 software.
Images were acquired using Zeiss LSM700 confocal microscope and Axio Imager.M2 compound microscope or with Discovery.V8 stereoscope and AxioCam MRc digital camera (Zeiss). Microscope objectives used were 40x 1.0 NA water objective or 25x 0.8 NA or 10x 0.3 NA. Images were exported as JPEG or TIFF files using ZEN 2009 LE software (Zeiss) and figures were assembled using Adobe Photoshop CS4 software.
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