Zebrafish experiments were approved by the UCD Animal Research Ethics Committee and the Healthcare Products Regulatory Authority (AE18982/P038). All methods were performed in accordance with the relevant guidelines and regulations. The transgenic Tg[dβh:MYCN-EGFP] zebrafish line was a generous gift from Prof. A. Thomas Look (Dana-Farber Cancer Institute, Harvard Medical School)33 (link). Adult Tg[dβh:MYCN-EGFP] zebrafish (Danio rerio) were maintained on a 14 h light/10 h dark cycle at 28 °C; and monitored for fluorescent tumor masses every two weeks. Approximately 20% of fish develop the equivalent of human neuroblastoma by about 8 months of age33 (link). Drug treatments of zebrafish were performed as described earlier40 (link). Zebrafish developing tumors (age: 21 months; n = 3) were treated with YM155 and lapatinib every 72 hours for a total of seven days. Fish water was changed every 72 hours, and drugs were dissolved in the fish water. Zebrafish were anesthetized by 0.016% Tricaine prior to microscopic analysis. An Olympus SZX10 fluorescent stereo microscope equipped with an Olympus DP71 camera was used to capture images. The fluorescent area posterior to the gills, which corresponds to the region of neuroblastoma growth33 (link), was analysed using the ImageJ software (v1.44p).
Szx10 fluorescent stereo microscope
Manufactured by Olympus
The Olympus SZX10 is a fluorescent stereo microscope designed for a wide range of applications. It features high-performance optics and advanced illumination systems to provide clear, high-contrast images. The SZX10 is capable of magnifications from 6.3x to 63x, making it suitable for various tasks that require detailed observation and analysis.
Automatically generated - may contain errors
3 protocols using szx10 fluorescent stereo microscope
1
Zebrafish Neuroblastoma Drug Treatment
2
Microscopy-based Biofilm and Single Cell Analysis
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Movies and snapshots were acquired with an IX71 inverted fluorescence microscope (Olympus), IX83 inverted fluorescence microscope (Olympus) or DeltaVision microscopy imaging system (GE Healthcare). Biofilm movies were taken from the bottom with a 2.5x objective lens (MPLFLN 2.5x/0.08, Olympus) to film the whole biofilm or a 10x objective lens (UPLFLN 10x/0.3, Olympus) to zoom in on certain regions. Images were taken every 40 min. Single cell movies were taken with a 100x objective lens (UPLFLN 100x/1.3, Olympus) while growing 10-fold diluted cell cultures on an MSgg pad at 30°C. Top-view of biofilms was acquired by a Retiga 2000R digital camera (QImaging) via an SZX10 fluorescent stereomicroscope (Olympus).
3
Transgenic Zebrafish Model of Neuroblastoma
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The transgenic Tg[dβh:MYCN-EGFP] zebrafish line was a generous gift from Prof. A. Thomas Look (Dana-Farber Cancer Institute, Harvard Medical School) (23) . Zebrafish experiments were approved by the UCD Animal Research Ethics Committee and the Healthcare Products Regulatory Authority (HRPA; AE18982/P038).
Adult Tg[dβh:MYCN-EGFP] zebrafish (Danio rerio) were maintained at 28°C on a 14 h light/10 h dark cycle and monitored for fluorescent tumor masses every two weeks. Approximately 20% of fish develop tumors by ~8 months (23) . Zebrafish developing the fish-equivalent of human neuroblastoma (age: 15 months; n=4) were treated every 48 h with JNK inhibitor II (5 μM) or DMSO control for a total of two weeks. Variance within tumor size for all fish at pre-treatment was <5%. Zebrafish were anesthetized by 0.016% Tricaine prior to microscopic analysis. An Olympus SZX10 fluorescent stereo microscope equipped with an Olympus DP71 camera was used to capture images. The fluorescent region posterior to the gills, which corresponds to the region of neuroblastoma growth (23) , was analyzed using ImageJ software (v1.44p).
Adult Tg[dβh:MYCN-EGFP] zebrafish (Danio rerio) were maintained at 28°C on a 14 h light/10 h dark cycle and monitored for fluorescent tumor masses every two weeks. Approximately 20% of fish develop tumors by ~8 months (23) . Zebrafish developing the fish-equivalent of human neuroblastoma (age: 15 months; n=4) were treated every 48 h with JNK inhibitor II (5 μM) or DMSO control for a total of two weeks. Variance within tumor size for all fish at pre-treatment was <5%. Zebrafish were anesthetized by 0.016% Tricaine prior to microscopic analysis. An Olympus SZX10 fluorescent stereo microscope equipped with an Olympus DP71 camera was used to capture images. The fluorescent region posterior to the gills, which corresponds to the region of neuroblastoma growth (23) , was analyzed using ImageJ software (v1.44p).
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