Transgenic fluorescent embryos were embedded in 1% agarose in a glass‐bottom dish. Immunofluorescence double staining was performed with chicken‐anti‐GFP (1:400; Life Technologies) and rabbit‐anti‐phospho‐histone3 (pH3) antibodies (1:250; Abcam). We used AlexaFluor488‐conjugated anti‐chicken (1:1,000; Life Technologies) and AlexaFluor594‐conjugated anti‐rabbit (1:1,000; Life Technologies) secondary antibodies to reveal primary antibodies. Confocal imaging was performed using a Nikon inverted A1r spectral.
Inverted a1r spectral
Manufactured by Nikon
The Inverted A1r spectral is a specialized laboratory equipment designed for advanced microscopy applications. It functions as an inverted fluorescence microscope, allowing for the observation and analysis of samples from the bottom up. The core function of this instrument is to provide high-resolution, high-sensitivity imaging capabilities for a wide range of biological and material science research applications.
Automatically generated - may contain errors
Lab products found in correlation
Chicken anti gfp, by Thermo Fisher Scientific (2 mentions)
Alexa fluor 594 conjugated anti rabbit, by Thermo Fisher Scientific (2 mentions)
Alexafluor488 conjugated anti chicken, by Thermo Fisher Scientific (2 mentions)
Rabbit anti phospho histone3 ph3 antibodies, by Abcam (1 mentions)
Gtx127342, by GeneTex (1 mentions)
Tricaine, by Merck Group (1 mentions)
4 protocols using inverted a1r spectral
1
Fluorescent Embryo Imaging with IF Staining
2
Imaging DNA Damage in Transgenic Embryos
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Transgenic fluorescent embryos were embedded in 1% agarose dissolved in 1xE3 in a glass-bottom dish. For immunofluorescence, double staining was performed using chicken-anti-GFP (1:500; Life Technologies), rabbit-anti-H2A.X (phospho Ser139) (1:500; GTX127342; Genetex) rabbit, and pH3 (1:250; Abcam) antibodies. We used AlexaFluor488-conjugated anti-chicken (1:1,000; Life Technologies) and AlexaFluor594-conjugated anti-rabbit (1:1,000; Life Technologies) secondary antibodies to reveal primary antibodies. Confocal imaging was performed using a Nikon inverted A1r spectral.
3
Time-Lapse Imaging of Hematopoiesis
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For time‐lapse imaging, Tg(cmyb:GFP) embryos were anesthetized with 0.03% Tricaine (Sigma) and embedded in 1% agarose in a 60 mm glass‐bottom dish. Embryos were imaged at 28.5 °C with a confocal Nikon inverted A1r spectral. Scanning with 20× water immersion objective, z‐stacks were acquired with a step‐size of 7 μm within an interval of 10 min for 6 h in the control‐ and slco2b1‐morphants starting at ∼54hpf. The analysis of cmyb:GFP+ cells in all experiments was performed using IMARIS image software.
4
Confocal Imaging of Live Anesthetized Embryos
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Live (anaesthetized) embryos were embedded in 0.5% agarose supplied with 0.07 mg per mL Tricaine in E3 in a glass-bottom dish. Confocal imaging was performed using a Nikon inverted A1r spectral. For time-lapse experiments, embryos were imaged in a heated chamber at 28°C overnight. The larvae were kept anaesthetized for the duration of the experiment (several hours to overnight). Images were analyzed and the contrast and brightness equally adjusted using the FIJI (ImageJ) software.
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