For high‐speed live imaging, Xenopus embryos injected with ANKRD55‐GFP and mCherry‐CLUAP1 mRNA were anaesthetized with 0.005% benzocaine at stage 26. High‐speed in vivo imaging was acquired on a Nikon Eclipse Ti confocal microscope with a 63×/1.4 oil immersion objective at 0.267 s per frame. Kymographs were calculated using Fiji (Schindelin et al, 2012 ). Confocal images were collected with an LSM700 inverted confocal microscope (Carl Zeiss) with a Plan‐APOCHROMAT 63×/1.4 oil immersion objective. Bright field images were collected using a Zeiss Axio Zoom V16 stereo microscope with Carl Zeiss Axiocam HRc color microscope camera. Neural tube closure quantification was performed using Fiji. A two‐sample Kolmogorov–Smirnov test was used to compare distributions of control, morphant, and rescue embryos. Sample sizes were selected sufficient to determine moderate effects. Embryos were randomly selected from multiple clutches, and cells were randomly selected from individual embryos. No blinding to treatment was employed.
Axiocam hrc color
Manufactured by Zeiss
The Axiocam HRc color is a high-resolution color camera designed for microscopy applications. It features a 12.5 megapixel CMOS sensor capable of capturing detailed images at high resolutions. The camera is compatible with a wide range of Zeiss microscopes and provides fast data transfer through a USB 3.0 interface.
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2 protocols using axiocam hrc color
1
Live Imaging of Neural Tube Closure
2
Metastatic Spread and Necrosis Evaluation
Check if the same lab product or an alternative is used in the 5 most similar protocols
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To compare the level of metastasis and associated necrosis between biopsied and non-biopsied mice, a total of 84 mice in four cohorts underwent the metastasis model described above. Cohort size was designed to allow manageable numbers of mice and enable consistent treatment and surgical handling. Seven days after the tumor excision, each cohort of mice consisting, on average, of biopsy (n = 10) and non-biopsy (n = 11) mice was sacrificed. The draining lymph nodes and lungs were harvested, formalin fixed, and paraffin embedded with each block assigned an anonymizing numerical identifier to allow unbiased evaluation. The anonymized lungs, lymph nodes, and tumors were serial sectioned into 5-μm thin tissue sections and stained with hematoxylin and eosin (H&E) for histologic evaluation. Imaging was performed using a Zeiss Axiocam HRC Color, mounted on a Zeiss Stemi. The percentage of metastatic (lung and lymph node) and necrotic (tumor) tissue areas relative to total section area was established using an overlay grid imposed on images captured from H&E-stained tissue sections.
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