爪蟾核心实验技术与操作规程,供科研人员参考。Core Xenopus protocols and SOPs for reference.
基于 Nieuwkoop–Faber 标准的高清彩色分期图,涵盖受精卵至自由游泳蝌蚪各阶段形态特征。High-resolution NF stage chart from fertilised egg to free-swimming tadpole, with key morphological features.
动物筛选、低温麻醉、手术取卵、OR2 保存液配方与胶原酶消化分离流程。Animal selection, cold anaesthesia, surgery, OR2 solution, and collagenase digestion protocol.
ND96 培养液配方、胶原酶消化(1 mg/mL,22℃)、Dumont V–VI 期筛选与质控标准。ND96 medium, collagenase digestion (1 mg/mL, 22 ℃), Dumont V–VI selection and QC criteria.
SCE 原理、微电极制备、电脉冲参数(20 V, τ=70 ms)与故障排除。SCE principle, micropipette prep, pulse parameters and troubleshooting.
脑室注射 75–125 nL 质粒 DNA、铂电极 30–50 V、单侧 / 双侧靶向转染。Ventricular injection 75–125 nL plasmid, Pt electrode 30–50 V, unilateral/bilateral targeting.
I–VI 期特征、直径与各类实验的选用建议。Stage I–VI features and selection for each assay.
cRNA 准备、注射量、针径与功能检测。cRNA prep, dose, needle and functional readout.
设备、针尖校准、注射位置与恢复培养。Equipment, calibration, site and recovery.
徒手持握法、鱼网固定法与动物福利要点。Hand-holding, net restraint and welfare notes.
显微操作、细胞追踪、移植与基因功能研究。Micromanipulation, tracing, grafting and gene function.
原理、设计、用量、对照与脱靶验证。Principle, design, dose, controls and validation.
Steinberg's、MBS-H、PBS 配方与 MS-222 麻醉。Steinberg's/MBS-H/PBS recipes and MS-222.
电极充填(3M KCl)、ND96 浴槽液配方、钳制步骤与常见问题排查。Electrode fill (3M KCl), ND96 bath solution, clamping procedure and troubleshooting.