爪蟾卵母细胞相关技术培训服务Xenopus oocyte technology training

爪蟾资源中心致力于帮助科研人员快速建立和掌握爪蟾实验技术体系。中心提供六大模块的系统培训课程，涵盖从动物养殖管理到 TEVC 电生理记录的全流程技术，采用小班制手把手指导，确保学员能够在本实验室独立开展实验。The Xenopus Resource Center helps researchers rapidly establish and master Xenopus experimental techniques. We offer six training modules covering the complete workflow from animal husbandry to TEVC electrophysiology. Small-group, hands-on instruction ensures participants can independently run experiments in their own labs.

培训体系特色Training program highlights

小班授课：每期 1–4 人，确保每位学员获得充分的实操时间；Small groups: 1–4 participants per session, ensuring ample hands-on time;
模块化课程：可根据需求选择单一或组合模块，灵活定制培训内容；Modular curriculum: select individual or combined modules to match your needs;
标准化操作规范：所有操作均按中心制定的 SOP 进行，学员可直接带走书面流程；Standardized SOPs: all procedures follow center-developed SOPs; participants take home written protocols;
长期技术支持：培训结束后提供 3 个月线上技术咨询支持；Long-term support: 3 months of online technical consultations after training;
真实实验材料：使用中心自养爪蟾提供新鲜卵母细胞与胚胎进行实训。Real materials: fresh oocytes and embryos from our own facility used in all practical sessions.

六大培训模块Six training modules

系统掌握爪蟾核心实验技术Systematically master core Xenopus techniques

模块 01Module 01

卵母细胞分离与制备Oocyte isolation and preparation

成体爪蟾手术麻醉与取卵操作规范Anaesthesia and surgical ovariectomy
胶原酶消化（I 型，1 mg/mL，22℃）分离卵泡细胞Collagenase I digestion (1 mg/mL, 22 °C) for follicle removal
Dumont I–VI 期卵母细胞的形态鉴别与分级筛选Morphological identification and grading of Dumont stage I–VI oocytes
ND96 培养液配制与卵母细胞日常维护ND96 medium preparation and daily oocyte maintenance
卵母细胞活力评估与质控标准Oocyte viability assessment and QC criteria

模块 02Module 02

分子克隆与载体构建Molecular cloning and vector construction

适合爪蟾表达的载体选择（pGEMHE、pCS2+ 等）Expression vector selection (pGEMHE, pCS2+, etc.)
酶切 / 连接、同源重组（Gibson Assembly / In-Fusion）策略Restriction-ligation, Gibson Assembly / In-Fusion recombination
目的基因优化（密码子优化、Kozak 序列）Target gene optimization (codon optimization, Kozak sequence)
N / C 端标签（FLAG / GFP / HA / mCherry）插入设计N/C-terminal tag insertion design (FLAG/GFP/HA/mCherry)
质粒验证（菌落 PCR、测序）Plasmid verification (colony PCR, sequencing)

模块 03Module 03

cRNA 体外合成与显微注射cRNA in-vitro synthesis and microinjection

质粒线性化（限制酶选择与纯化）Plasmid linearization (restriction enzyme selection and purification)
体外转录体系（T3/T7 RNA 聚合酶，5' 帽修饰，3' Poly-A）IVT system (T3/T7 polymerase, 5' cap, 3' poly-A)
cRNA 浓度与质量检测（凝胶电泳 + 分光光度计）cRNA quality and concentration (gel + spectrophotometry)
显微注射仪器操作（Drummond Nanoject II / Nanoliter 2010）Microinjection instrument operation (Drummond Nanoject II / Nanoliter 2010)
注射量校准（50–70 nL / 个）与成功率优化Volume calibration (50–70 nL/cell) and yield optimization
注射后培养与表达时间优化（1–4 天）Post-injection culture and expression timing (1–4 days)

模块 04Module 04

TEVC 双电极电压钳记录TEVC two-electrode voltage clamp recording

TEVC 基本原理与仪器构成（放大器、浴槽、虚地电极）TEVC principle and instrument setup (amplifier, bath chamber, virtual ground)
玻璃微电极拉制（参数设置、阻抗 0.5–2 MΩ）与充填（3 M KCl）Glass microelectrode pulling (parameters, 0.5–2 MΩ) and filling (3 M KCl)
卵母细胞固定与电极穿刺（电压 + 电流双电极定位）Oocyte fixation and electrode impalement (voltage + current electrode)
标准电压方案设计（阶梯 / 斜坡 / 持续电压）Standard voltage protocol design (step/ramp/sustained voltage)
I-V 曲线记录与漏电流校正I-V curve recording and leak-current subtraction
数据采集软件（Clampex / pCLAMP）与分析（Clampfit）操作Data acquisition (Clampex/pCLAMP) and analysis (Clampfit) operation
常见问题排查（噪声、封闭失败、细胞死亡）Troubleshooting (noise, poor seal, cell death)

模块 05Module 05

胚胎操作与发育表型分析Embryo manipulation and developmental phenotyping

激素诱导产卵（HCG / PMSG 注射方案）Hormone-induced spawning (HCG/PMSG injection protocols)
体外受精与胚胎去卵壳膜操作In-vitro fertilization and jelly-coat removal
早期胚胎特定卵裂球注射（基因过表达、Morpholino 敲低）Targeted blastomere injection for overexpression or Morpholino knockdown
NF 分期与表型观察记录NF staging and phenotype documentation
整体原位杂交（WISH）基础操作Whole-mount in-situ hybridization (WISH) basics
免疫荧光标记与共聚焦成像Immunofluorescence labeling and confocal imaging

模块 06Module 06

养殖管理与实验动物伦理规范Husbandry management and laboratory animal ethics

爪蟾饲养环境标准（水质、温度、光照、密度）Housing standards (water quality, temperature, lighting, density)
饲料种类与投喂频率Diet types and feeding frequency
常见疾病识别与处理（红腿病、真菌感染）Common disease recognition and treatment (red-leg syndrome, fungal infection)
MS-222 麻醉操作与安乐死规范MS-222 anaesthesia and euthanasia protocols
实验动物使用许可证申请与 IACUC 申报要点Animal use permit application and IACUC protocol submission
爪蟾入侵物种风险管控（废弃动物处置规范）Invasive species risk management (disposal of unwanted animals)

培训安排Training schedule

灵活定制，按需组合Flexible customization, modular combination

培训形式Format	时长Duration	内容Content	人数Group size
单模块集中培训Single-module intensive	1–2 天	选定模块理论 + 操作Selected module theory + practice	1–4 人
基础组合（模块 01+02+03）Foundation combo (01+02+03)	3–4 天	从动物管理到 cRNA 注射完整流程Husbandry to cRNA injection full workflow	1–4 人
电生理组合（模块 01+02+03+04）Electrophysiology combo (01+02+03+04)	5–6 天	cRNA 制备到 TEVC 记录完整技术链cRNA to TEVC complete technical chain	1–3 人
全程班（全部六模块）Full program (all six modules)	8–10 天	爪蟾实验平台搭建全流程Complete Xenopus lab platform setup	1–2 人
线上理论课（远程）Online theory (remote)	半天 / 模块	概念讲解 + 视频演示（不含实操）Concept lecture + video demo (no practicals)	不限Unlimited

快速建立你的爪蟾实验能力Rapidly build your Xenopus experimental capacity

选择培训模块，告知我们你的实验目标，我们为你定制培训方案与时间安排。Choose your modules, describe your experimental goals — we design a custom training plan and schedule.

预约培训Book training 查阅技术资料Technical resources