非洲爪蟾的注射规范操作Standard injection handling

1. 目的Purpose

规范非洲爪蟾在实验过程中的操作方法，最大限度减少动物应激与损伤，防止感染传播，保障动物福利与实验安全。Standardise handling of Xenopus during experiments to minimise stress and injury, prevent pathogen transfer, and ensure animal welfare and laboratory safety.

2. 操作前准备Preparation

• 操作者应彻底洗净双手，确保无刺激性化学物残留（如肥皂、酒精、消毒剂等）。Wash hands thoroughly — remove all irritant residues (soap, alcohol, disinfectants).
• 禁止佩戴乳胶、丁腈或其他类型手套直接操作动物；手套材料对爪蟾皮肤及黏膜有刺激性。Do not handle frogs with latex, nitrile or any other gloves — glove materials irritate frog skin and mucous membranes.
• 备好注射工具、配套缓冲液、注射平台与鱼网等操作辅助物品，注射针提前校准体积。Prepare injection tools, buffers, injection platform and net; calibrate injection volume in advance.

3. 徒手持握法Hand-holding technique

• 使用裸手（不佩戴手套）直接操作动物。Handle with bare hands (no gloves).
• 左手温和持握爪蟾，掌心托住其腹面，拇指和食指轻轻支撑背部两侧。Cup the frog in the left hand, palm under the belly, thumb and index finger gently supporting the sides of the back.
• 用手指轻遮眼部，帮助动物安静，减少挣扎与应激。Lightly cover the eyes to calm the animal and reduce struggling.
• 禁止施加挤压性压力，避免内脏压迫、肺破裂或皮肤损伤。Never squeeze — this can compress viscera, rupture lungs or damage skin.
• 动物平稳后，右手进行背部皮下淋巴囊注射或腹腔注射等操作，注射后温和释放。Once the animal is calm, inject with the right hand (dorsal subcutaneous lymph sac or intraperitoneal); release gently afterwards.

4. 鱼网固定法（推荐用于难以安抚的个体）Net restraint (for uncooperative individuals)

• 使用清洁的细网鱼网（网眼细密）捞取个体。Use a clean fine-mesh net to capture the animal.
• 将爪蟾轻轻置于网中，将网口轻度卷合，使其保持自然蜷缩状态，不得强行收紧。Place the frog in the net and gently roll the opening closed — do not overtighten.
• 用左手固定网体底部，确保爪蟾不滑出；右手在透网空间或尾部裸露区域进行注射。Hold the net base with the left hand; inject through the mesh gaps or at the exposed rear.
• 操作完成后缓慢松网，将动物温和释放或转入恢复容器，切勿用力摇甩。Release the net slowly; transfer to a recovery container gently — do not shake or drop.

5. 注意事项与生物安全Safety & biosafety

• 切勿强制按压动物或在其挣扎时注射；强行操作可导致针头折断或动物内伤。Never inject a struggling animal — forceful injection can break needles or cause internal injury.
• 所有注射设备应保持无菌或经适当消毒处理，防止针头污染后续样本。Keep all injection equipment sterile or properly disinfected to avoid sample cross-contamination.
• 操作结束后彻底清洗双手，防范Salmonella spp. 等潜在病原体的交叉传播，尤其在接触其他水生动物或食物之前。Wash hands thoroughly after handling — prevent cross-contamination from Salmonella spp. and other potential pathogens, particularly before touching other animals or food.
• 废弃注射器与针头按医疗废物处理；废弃缓冲液与生物材料按机构规定处置。Dispose of syringes and needles as medical waste; discard buffers and biological material per institutional guidelines.

参考文献References

1. Wu M & Gerhart J (1991) Raising Xenopus in the laboratory. Methods Cell Biol 36:3–18.
2. Green SL (2010) The Laboratory Xenopus sp. CRC Press, Boca Raton.
3. Gurdon JB & Hopwood N (2000) The introduction of Xenopus laevis into developmental biology. Int J Dev Biol 44:43–50.