计算溶液所需的质量、体积或浓度。
动物灌注固定液 (4% PFA)
有货
为什么选择此级别
BioReagent,即用型 级 ，适用于对基线干扰要求严格的色谱和分析工作流程。
储存与运输
室温。常规运输 。请查阅批次 COA 获取详细规格。
质量文档
SDS、COA、产品数据表及规格说明书均可下载。可通过批号查询获取批次 COA。
文献证明
在色谱分析、有机合成和交叉偶联反应领域已被 0 篇同行评审文献引用。
概述
灌注固定是解剖学和神经科学等领域中一种用于制备高质量组织样本的关键技术。它指的是在动物 (通常是小鼠、大鼠等实验动物) 经深度麻醉后，通过其循环系统 (通常是心血管系统) 快速注入固定液，从而迅速、均匀地终止细胞内所有生化活动，将组织结构和生物大分子 “冻结” 在生命状态的某一瞬间。其核心在于利用动物自身的血管网络作为输送管道，使固定液能到达全身每一个角落。
与简单的将组织浸泡在固定液中 (immersion fixation，浸泡固定) 相比，灌注固定具有无可比拟的优势：
①速度极快，固定效果极佳；
②形态结构保存完好；
③适用于深部组织；
④减少可溶性物质的流失；
⑤为后续实验提供基础。
动物灌注固定液 (4% PFA) 主要由多聚甲醛、磷酸盐等组成，pH 为 7.4 左右，经过滤处理。本产品适合于绝大多数动物的灌注固定。该产品固定均匀，组织收缩轻微。
该试剂仅用于科研领域，不适用于临床诊断或其他用途。
操作步骤 (以大鼠 / 小鼠经心脏灌注为例):
1、麻醉：
使用过量麻醉剂 (如戊巴比妥钠) 腹腔注射，确保动物深度麻醉 (以疼痛刺激无反应为准)。
2、固定与开胸：
(1) 将动物仰卧位固定在冰盘上的操作板上 (低温可延缓组织降解)。
(2) 用酒精消毒胸腹部皮肤。
(3) 用剪刀沿中线剪开腹部皮肤和肌肉，向上剪开胸腔，暴露肋骨。小心剪开膈肌，然后向两侧剪开肋骨，充分暴露心脏。
3、插管：
(1) 用镊子轻轻提起心包，暴露心脏。
(2) 将 perfusion 针头从左心室心尖部插入，朝向主动脉弓方向，并轻轻推进至主动脉内。
(3) 用动脉夹或缝合线将针头固定在心脏内，防止滑脱。
(4) 立即用手术剪剪开右心耳，为灌流液创建流出通道。
4、灌注：
(1) 打开装有 4℃生理盐水的吊瓶开关，以快速流速 (小鼠约 10‑20 mL/min，大鼠约 50‑100 mL/min) 进行灌注。肝脏颜色迅速变白，右心耳流出液从鲜红色逐渐变为无色。通常灌注量约为动物体重的 2 倍 (如 200 g 大鼠灌注 400 mL)。动物四肢、尾巴和肝脏会剧烈抽搐后变得僵硬。
(2) 当流出的液体基本无色时，迅速切换到装有 4% PFA 的吊瓶。改为慢速灌注 (小鼠约 5‑10 mL/min，大鼠约 20‑30 mL/min)。动物身体会再次出现缓慢的全身性强直 (四肢、尾巴、颈部僵硬)，这是固定成功的标志。灌注量同样约为动物体重的 2 倍。
5、取材与后固定：
(1) 关闭灌注系统，取出针头。小心取出目标组织 (如脑、脊髓)。
(2) 将组织块立即放入新鲜冰冷的 4% PFA 中，4℃下进行后固定。后固定时间至关重要：
对于常规光镜样本：6‑24 小时 (视组织块大小而定)。
时间过短：固定不充分。
时间过长：组织过度交联，变脆，并影响后续抗体染色 (抗原遮蔽)。
(3) 后固定完成后，将组织转移至蔗糖‑PBS 溶液 (如 20%‑30%) 中，4℃下沉降直至组织完全沉底，然后进行冷冻切片或长期保存。
注意事项:
1、动物灌注固定液对人体有一定的损害，请在通风好的环境下小心操作，避免吸入。
2、组织取材的厚度不同，固定时间也不同，对组织恰当的选材有利于固定液的渗透。
3、固定液的容量应足够，一般固定液与组织块的体积比率应大于 10:1。
4、温度对固定的影响很明显，提高温度可以加速固定作用，但温度不宜过高。
5、取出新鲜组织后应及时固定，无法及时固定时应保存于生理盐水中及时送检。
6、为了您的安全和健康，请穿实验服并戴一次性手套操作。
7、试剂开封后请尽快使用，以防影响后续实验效果。
Perfusion fixation is a key technique for preparing high‑quality tissue samples in anatomy, neuroscience and other disciplines. After experimental animals (commonly mice and rats) are deeply anesthetized, fixative is rapidly infused via the circulatory system (cardiovascular system in most cases). This promptly and uniformly terminates all intracellular biochemical activities and “freezes” tissue architecture and biomacromolecules at a certain moment of the living state. Its core principle is to utilize the animal’s own vascular network as delivery channels, enabling the fixative to reach every part of the body.
Compared with simple immersion fixation, perfusion fixation has unparalleled advantages:
① Extremely fast rate with excellent fixation performance;
② Well‑preserved tissue morphology and structure;
③ Applicable to deep‑layer tissues;
④ Reduced loss of soluble substances;
⑤ Provides reliable foundations for subsequent experiments.
Perfusion Fixative for Animal (4% PFA) is mainly composed of paraformaldehyde and phosphate, with pH approximately 7.4 and pre‑filtration treatment. It is suitable for perfusion fixation of most laboratory animals, featuring uniform fixation and minimal tissue shrinkage.
This reagent is for research use only and is not intended for clinical diagnosis or other applications.
Operating Procedures (Transcardiac Perfusion in Rats / Mice for Reference Only)
1. Anesthesia
Inject an overdose of anesthetic (e.g. sodium pentobarbital) intraperitoneally to achieve deep anesthesia, confirmed by absence of response to painful stimuli.
2. Restraint and Thoracotomy
(1) Secure the animal in supine position on an operating plate placed over an ice tray (low temperature slows tissue degradation).
(2) Disinfect the skin of the thorax and abdomen with alcohol.
(3) Cut the abdominal skin and muscle along the midline, open the thoracic cavity upwards to expose the ribs. Gently incise the diaphragm, cut the ribs bilaterally, and fully expose the heart.
3. Cannulation
(1) Gently lift the pericardium with forceps to expose the heart.
(2) Insert the perfusion needle through the apex of the left ventricle towards the aortic arch, and advance gently into the aorta.
(3) Secure the needle within the heart using an arterial clip or suture to prevent dislodgement.
(4) Immediately cut the right auricle with surgical scissors to create an outflow channel for perfusate.
4. Perfusion
(1) Turn on the drip bottle containing 4 ℃ normal saline and perfuse at high flow rate (~10‑20 mL/min for mice; ~50‑100 mL/min for rats). The liver rapidly blanches, and effluent from the right auricle changes from bright red to nearly colorless. The perfusion volume is generally twice the animal’s body weight (e.g. 400 mL for a 200 g rat). Limbs, tail and liver will twitch violently and then stiffen.
(2) When the outflow becomes almost colorless, swiftly switch to the drip bottle with 4% PFA. Perfuse at reduced flow rate (~5‑10 mL/min for mice; ~20‑30 mL/min for rats). Slow generalized tonic contraction (stiff limbs, tail and neck) occurs in the animal, indicating successful fixation. The perfusion volume is also approximately twice the animal’s body weight.
5. Tissue Harvesting and Post‑fixation
(1) Shut down the perfusion system and remove the needle. Carefully dissect target tissues (e.g. brain, spinal cord).
(2) Immediately place tissue blocks into fresh ice‑cold 4% PFA for post‑fixation at 4 ℃. Post‑fixation duration is critical:
For conventional light‑microscopy samples: 6‑24 h depending on tissue block size.
Insufficient duration leads to incomplete fixation.
Excessive duration causes biomacromolecule over‑crosslinking, tissue brittleness and impaired antibody staining (antigen masking).
(3) Upon completion of post‑fixation, transfer tissues into sucrose‑PBS solution (e.g. 20%‑30%). Keep at 4 ℃ until tissues sink completely to the bottom, then proceed to frozen‑section preparation or long‑term storage.
Precautions
1. Perfusion Fixative for Animal is hazardous to human body. Operate carefully in a well‑ventilated environment and avoid inhalation.
2. Fixation time varies with specimen thickness; proper sampling facilitates fixative penetration.
3. Sufficient fixative volume is required; generally the volume ratio of fixative to tissue block shall be greater than 10:1.
4. Temperature exerts remarkable influence on fixation. Elevated temperature accelerates fixation, yet excessive temperature should be avoided.
5. Fresh tissues should be fixed promptly after dissection. If immediate fixation is unavailable, store tissues in normal saline for timely processing.
6. Wear a lab coat and disposable gloves for your safety and health during operation.
7. Use the reagent as soon as possible after opening to avoid compromised experimental performance.
规格
规格或纯度
即用型, BioReagent
稳定性与储存
Store at RT long term (12 months).
英文名称
Perfusion Fixative for Animal (4% PFA)
储存条件
室温
运输条件
常规运输
产品线
生物产品
名称和识别符
分子类型
生物试剂/缓冲液
技术文档
高级数据
产品属性
| pH | 7.4 |
|---|
质检证书（CoA,COO,BSE/TSE 和分析图谱)
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通过匹配包装上的批号来查找并下载产品的 COA，每批产品都进行了严格的验证，您可放心使用！
| 批号(Lot Number) | 证书类型 | 货号 |
|---|---|---|
 ZJ26F1040063 |
分析证书 | P1529305 |
技术文档和文章
溶液计算器
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