计算溶液所需的质量、体积或浓度。
快速高尔基染色试剂盒
有货
概述
Golgi‑Cox 浸染法是研究神经元与胶质细胞正常及异常形态的经典有效方法之一。采用高尔基染色法处理神经组织时，仅能标记少量神经元，且方法稳定性、重复性较差，但该技术实现了人类首次对神经纤维的清晰观测。传统 Golgi 染色操作耗时、结果不稳定，限制了该方法的广泛应用。
本试剂盒无需多聚甲醛灌注，也无需频繁更换浸染液，大幅优化并简化 Golgi‑Cox 实验流程，提升实验重复性与结果可靠性。试剂盒经过大量验证，适用于多种动物脑组织及人脑组织样本。 本试剂仅用于科研领域，不适用于临床诊断或其他用途。
组分与储存：
| 组分货号 | 组分名称 | 物理外观 | 25T | 50T | 储存 |
| R1539901A | 溶液A | 液体 | 125mL | 250mL | 室温 |
| R1539901B | 溶液B (2×) | 液体 | 60mL | 125mL | 室温 |
| R1539901C | 溶液C | 液体 | 125mL | 250mL | 室温 |
| R1539901D | 染色反应瓶 | 固体 | 2EA | 2EA | 室温 |
| R1539901E | 天然毛画笔 | 固体 | 1EA | 1EA | 室温 |
| R1539901F | 塑料镊子 | 固体 | 1EA | 1EA | 室温 |
自备组分：
1. 双蒸水或 Milli-Q 水
2. 振动切片机
3. 光学显微镜
4. 组织学耗材和试剂： 防脱载玻片、盖玻片、玻片架、染色罐 、六孔板、塑料吸管、乙醇、二甲苯
安全操作注意事项：
1.该试剂盒仅用于体外研究，不能用于诊断或其它应用。
2. 试剂盒所包含有试剂是有毒的，吸入或接触皮肤是有害的，如果吞咽可能是致命的。不要用嘴吸。避免吸入和接触皮肤和眼睛。如果接触，立即用大量的水冲洗并求助医生。
3. 在化学通风橱下完成实验。操作这些试剂时须穿戴合适的防护服，手套和眼睛和面部防护罩，完成实验之后把手彻底冲洗干净。
试剂准备：
溶液B (2×)加入等体积去离子水进行稀释。
注意：溶液C有强烈刺激气味，请在通风橱内操作完成！
组织制备 (请仔细阅读)：
1. 杀死动物之前对实验动物进行深度麻醉。应尽快地从颅骨中取出动物的脑（或死去病人的脑），但操作时必须非常小心以免损伤或压迫组织。
注意：除非完全必要，不要对动物进行灌注。如果确实需要对动物进行灌注（用4%的多聚甲醛处理不要超过 5 分钟），一定不要对组织进行后固定处理。
体积较大的脑部样本包括大鼠脑部应该用锋利的刀片切成大约 10 mm 厚的块状。
2. 用双蒸水或 Milli-Q 水快速冲掉组织表面的血液。
3. 把组织浸泡在由溶液A的浸渍液中，室温下黑暗保存10天*。
*对于大多数情况浸泡10天是足够的。然而，不同的组织类型及组织的大小不同可能需要延长或缩短浸泡时间来达到最佳效果。对于每种类型的组织的浸泡时间应该通过试验获得，但是对于大多数组织浸泡 10天是足够的。注意，对于皮层等浅层脑组织可以缩短浸泡时间至7天，对于深部核团或大块组织可延长浸泡时间至14天，但是可能增加浅层组织的染色背景。
注意：溶液A接触皮肤是有毒的，如果吞咽可能是致命的。溶液B有易挥发，有刺激性味道，应该在化学通风橱中完成。当操作这些试剂时应穿戴防护服，手套和防护面具/眼镜。不要把溶液A 和B 的废弃物倒进水槽中。溶液 A 和 B 的废弃物需收集起来放至瓶子中，致电安全办公室或有执照的专业废物处理服务的部门处理。
每1 cm³待研究组织至少用 10 ml 浸泡液。用较少的浸泡液可能降低染色的灵敏性和可靠性。
4. 75%乙醇→95%乙醇→无水乙醇依次浸泡过夜脱水。
5. 室温条件下用振动切片机将组织切成 100-200 μm厚的薄片（进行切片之前在切片收集槽内放入适量去离子水保证组织块和切片浸没在水中）。用塑料吸管或毛笔将感兴趣的脑片转移至盛有去离子水的脑片回收皿中。为取得最佳结果，应尽快地完成切片，制备好的切片如果保存于脑片回收皿中，室温条件下最长可保存2 天。
6. 用移液枪吸弃脑片回收皿中的去离子水，加入适量溶液B，浸泡30 分钟（此时脑片变为黑灰色，为显色过程）。
7. 吸弃溶液B，加入适量去离子水轻柔漂洗5 分钟后弃去洗涤液，加入适量溶液C，浸泡10 分钟（此时脑片变为半透明并部分卷曲，为正常现象）。
8. 吸弃溶液C，加入适量去离子水轻柔漂洗10 分钟，75%乙醇→95%乙醇→无水乙醇依次浸泡30秒，进行脱水（此过程不可让切片变干）。
9. 使用毛笔（试剂盒提供）将无水乙醇中的脑片转移至阳离子包被的显微镜载玻片上，小心展平并晾干（晾干过程中可能出现部分脑片边缘部分卷曲，此时使用毛笔，小心再次展平，一旦乙醇完全挥发，脑片干透后呈现脆性，不可再次展平）。
10. 在二甲苯中透明，2次，每次1分钟，并且用树脂封片剂对盖玻片进行封片,显微镜明场下观察树突棘形态和数量。
11. 随机选择树突进行成像。拍照和统计依照盲法进行。树突棘密度的计算方法是从细胞体开始每10 μm树突一段，计数树突棘。树突棘的形态学根据之前描述的标准分类:“瘦长”形有长颈和小脑袋；“矮胖形”的有一个大脑袋，但没有脖子；“蘑菇”型有一个大脑袋，粗脖子。来自同一处理组的神经元的数值被合并在一起，计算平均值。
The Golgi‑Cox impregnation method is one of the most effective methods to study the normal and abnormal morphology of neurons and glial cells. When Golgi staining is applied to neural tissue, only a small number of neurons can be labeled, and the method exhibits poor stability and reproducibility. Nevertheless, this technique enabled the first clear observation of nerve fibers. Conventional Golgi staining is time-consuming and unreliable, which limits its widespread application.
This kit does not require paraformaldehyde perfusion or frequent replacement of impregnation solution. It greatly improves and simplifies the Golgi‑Cox protocol and enhances experimental reproducibility and reliability. Widely validated, it is suitable for brain tissues from various animal species and human brain tissue.
For research use only. Not for clinical diagnosis or other applications.
Contents & Storage
| Component No. | Component Name | Appearance | 25T | 50T | Storage |
| R1539901A | Solution A | Liquid | 125mL | 250mL | Room Temperature |
| R1539901B | Solution B (2×) | Liquid | 60mL | 125mL | Room Temperature |
| R1539901C | Solution C | Liquid | 125mL | 250mL | Room Temperature |
| R1539901D | Staining reaction vial | Solid | 2EA | 2EA | Room Temperature |
| R1539901E | Natural‑hair paintbrush | Solid | 1EA | 1EA | Room Temperature |
| R1539901F | Plastic forceps | Solid | 1EA | 1EA | Room Temperature |
Materials Required but Not Included
1.Double-distilled water or Milli-Q water
2.Vibratome
3.Light microscope
4.Histology consumables and reagents: adhesive-coated slides, coverslips, slide racks, staining jars, six-well plates, plastic pipettes, ethanol, xylene
Safety Precautions
1.This kit is for in vitro research use only and must not be used for diagnosis or other applications.
2.The reagents included in the kit are toxic. Inhalation or skin contact is harmful, and ingestion may be fatal. Do not mouth pipette. Avoid inhalation and contact with skin and eyes. In case of contact, immediately rinse with plenty of water and seek medical attention.
3.Perform the experiment in a chemical fume hood. Wear appropriate protective clothing, gloves, and eye and face protection when handling these reagents. Thoroughly wash hands after completing the experiment.
Reagent Preparation
Dilute Solution B (2×) by adding an equal volume of deionized water.
Note: Solution C has a strong irritating odor. Please operate in a fume hood!
Tissue Preparation (Please Read Carefully)
1.Deeply anesthetize the experimental animal before sacrificing it. Remove the animal brain (or the brain of a deceased patient) from the skull as quickly as possible, but handle with great care to avoid damaging or compressing the tissue.
Note: Do not perfuse the animal unless absolutely necessary. If perfusion is indeed required (treatment with 4% paraformaldehyde should not exceed 5 minutes), do not post-fix the tissue.
Larger brain samples, including rat brains, should be cut into blocks approximately 10 mm thick with a sharp blade.
2.Quickly rinse blood from the tissue surface with double-distilled water or Milli-Q water.
3.Immerse the tissue in the impregnation solution of Solution A and store in the dark at room temperature for 10 days*.
*In most cases, 10 days of immersion is sufficient. However, different tissue types and sizes may require longer or shorter immersion times to achieve optimal results. The immersion time for each tissue type should be determined experimentally, but 10 days is sufficient for most tissues. Note that for superficial brain tissues such as the cortex, the immersion time can be shortened to 7 days, while for deep nuclei or large tissue blocks, it can be extended to 14 days, but this may increase the staining background of superficial tissues.
Note: Solution A is toxic upon skin contact and may be fatal if swallowed. Solution B is volatile and has an irritating odor; it should be handled in a chemical fume hood. Wear protective clothing, gloves, and a protective mask/glasses when handling these reagents. Do not pour waste from Solutions A and B down the sink. Waste from Solutions A and B should be collected in a bottle, and the safety office or a licensed professional waste disposal service should be contacted to dispose of these materials.
Use at least 10 mL of impregnation solution per 1 cm³ of tissue to be studied. Using less impregnation solution may reduce the sensitivity and reliability of staining.
4.Dehydrate by sequential immersion in 75% ethanol → 95% ethanol → absolute ethanol overnight.
5.At room temperature, cut the tissue into 100–200 μm thick sections using a vibratome (before sectioning, place an appropriate amount of deionized water in the section collection bath to ensure the tissue block and sections are submerged in water). Transfer the brain sections of interest to a brain section recovery dish containing deionized water using a plastic pipette or a brush. For best results, sectioning should be completed as quickly as possible. Prepared sections can be stored in the brain section recovery dish at room temperature for up to 2 days.
6.Aspirate and discard the deionized water from the brain section recovery dish using a pipette, add an appropriate amount of Solution B, and immerse for 30 minutes (the brain sections turn black-gray at this point; this is the color development process).
7.Aspirate and discard Solution B, add an appropriate amount of deionized water, rinse gently for 5 minutes, then discard the wash solution. Add an appropriate amount of Solution C and immerse for 10 minutes (the brain sections become semi-transparent and partially curled at this point; this is normal).
8.Aspirate and discard Solution C, add an appropriate amount of deionized water, rinse gently for 10 minutes, then dehydrate by sequential immersion in 75% ethanol → 95% ethanol → absolute ethanol for 30 seconds each (do not allow the sections to dry during this process).
9.Transfer the brain sections from absolute ethanol to cation-coated microscope slides using a brush (provided in the kit), carefully flatten them, and air-dry (during drying, some edges of the brain sections may curl; use the brush to carefully flatten them again. Once the ethanol has completely evaporated and the brain sections are thoroughly dry, they become brittle and cannot be flattened again).
10.Clear in xylene, 2 times, 1 minute each, and mount with a resin mounting medium and a coverslip. Observe dendritic spine morphology and number under a bright-field microscope.
11.Randomly select dendrites for imaging. Photograph and quantify according to blind methods. Dendritic spine density is calculated by counting dendritic spines per 10 μm segment of dendrite starting from the cell body. The morphology of dendritic spines is classified according to previously described criteria: "thin" spines have a long neck and a small head; "stubby" spines have a large head but no neck; "mushroom" spines have a large head and a thick neck. Values from neurons in the same treatment group are pooled together to calculate the mean.
规格
别名
神经元高尔基染色试剂盒
英文别名
All-in-One Exosome Extraction & Purification Kit
规格或纯度
BioReagent, 用于显微镜
英文名称
Golgi‑Cox Stain Kit
样品类型
动物, 组织
储存与运输
储存条件
室温
运输条件
常规运输
稳定性与储存
Store at RT long term (24 months).
组分与储存
| 组分货号 | 组分名称 | 物理外观 | 25T | 50T | 储存 |
| R1539901A | 溶液A | 液体 | 125mL | 250mL | 室温 |
| R1539901B | 溶液B (2×) | 液体 | 60mL | 125mL | 室温 |
| R1539901C | 溶液C | 液体 | 125mL | 250mL | 室温 |
| R1539901D | 染色反应瓶 | 固体 | 2EA | 2EA | 室温 |
| R1539901E | 天然毛画笔 | 固体 | 1EA | 1EA | 室温 |
| R1539901F | 塑料镊子 | 固体 | 1EA | 1EA | 室温 |
高级数据
质检证书(CoA,COO,BSE/TSE 和分析图谱)
C of A & Other Certificates(BSE/TSE, COO):
输入批号以搜索分析图谱:
溶液计算器
产品仅提供给经过验证的企业、机构和合格的专业人员用于研发。不适用于人类、动物、诊断或治疗。
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