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Small sections of the tail (2 mm) were dissected from F0 embryos and then analysed to verify mutagenesis by the direct sequencing of PCR amplicons assay73.
The portal vein (diameter, 5 10 mm), supra- and infra-hepatic inferior vena cava (IVC) (diameter, 12 15 mm), and common bile duct (diameter, 1.5 3.0 mm) were dissected out.
From each toluidine blue-stained section, four rectangular regions of approximately 1×1.4 mm were dissected with a Leica SVS LMD System (Leica Microsystems).
Then, coronal sections of approximately 2 mm (interaural position 4.7 to 6.7 mm corresponding to Bregma −4.3 to −2.3 mm) were dissected from the cerebrum of adult rats and the corresponding region in younger animals.
The distal parts of the soleus and gastrocnemius muscles (5-8 × 10 mm) were dissected out.
Thin ventricular trabeculae (diameter <0.6 mm) were dissected with the help of a stereoscopic microscope.
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Thereafter, a rectangular scleral flap measuring 4 × 3 mm was dissected.
The NP was isolated after the outermost layer of the AF (about 0.5 mm) was dissected and discarded to prevent contamination with cells from surrounding ligaments.
An area of approximately 10 30 mm was dissected from each patient and isolated in 300 μl of lysis buffer (Gentra Systems Inc., Minneapolis, MN, USA).
After conjunctival incision at the 12 o´clock position, a superficial parabolic scleral flap measuring 5 × 5 mm was dissected and a smaller deep flap was created to allow access to Schlemm canal.
The septal coronary artery (4 6 mm) was dissected, cleared of adhering tissue and incubated with 20 μM Fluo-4 AM for 3 h at room temperature in the presence of 0.25% of the non-ionic detergent Pluronic F-127.
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