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Hito NeuronMyelinStain™ Kit is designed based on Luxol fast blue and Cresyl echt violet staining method. This kit is made in a ready-to-use format and offers high quality, rapid staining of myelin/myelinated axons and neurons on paraffin sectionsor frozen sections. It has been proven to be extremely reliable and sensitive for simultaneously demonstrating the morphological details of myelin fibers and neurons.
Myelin is a substance rich in lipids and proteins that forms a layer, the myelin sheath, around only the axon of a neuron and acts as insulation. It is essential for the proper functioning of the nervous system. Demyelination is the loss of the myelin sheath insulating the nerves. This impairs the conduction of signals in the affected nerves, causing impairment in sensation, movement, and cognition. Currently no cure exists for demyelinating diseases and myelin repair is an active research field. Hito NeuronMyelinStain™ Kit allows sensitive localization and visualization of the myelin fibers, thus offers a fast and reliable way to determine the extent of demyelination.Hito NeuronMyelinStain™ Kit has been tested extensively on the brains and spinal cord from several species of animals and it is a simple solution for your research. HitoNeuronMyelinStainKit Solution-1 is designed to contain Alcohol that is flammable. However, shipping hazardous materials internationally comes with high cost and long waiting time for customs clearance. This PreKit is designed for customers who can purchaseabsolute Alcohol in their own countries, and then add to the PreKit (Solution-1 Stock Solution) for fast and safe usage of the kit.
Kit Contents Standard Kit Solution-1 (Luxol Fast Blue Stock Solution) 10 ml Solution-2 (Lithium Carbonate Solution 5xConcentrate) 250 ml Solution-3 (Cresyl Echt Violet Solution) 30 ml Dropper Bottle 1 Staining Jars 3 User Manual and MSDS 1 Before using Hito NeuronMyelinStain™ Kit, please make sure you have the following Required Equipment / Materials in your lab (not included in the kit):Cryostat or Microtome and light microscopeParaffin embedding equipment(or paraffin sections)Hito Bouin’s Plus Solution (Cat# HTSH0102)Dry ice, isopentane, O.C.T. compound (for frozen sections), ethanol, xylene, 4% PFA (Hito Cat# HTSHS0102), double distilled or deionized waterSlides and coverslipsStaining jars for slides washResinous mounting medium
Hito NeuronMyelinStain PreKitManual and MSDS- Additional Information
Additional Information
SKU HTKNS1225NH - Reviews
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但在干冰或-70℃冰箱中保存的细胞，活力下降很快，当然细胞与细胞之间敏感性不同。有实验验证保存在-70℃冰箱中的细胞6个月后活力损失超过50%。ATCC证明在干冰中人类细胞4个月活力损失殆尽，小鼠的细胞可到6个月。
2×miRcute。
伤,有必要使用含有DMSO冷冻保护剂。 DMSO能够快速穿透细胞膜进入细胞中,降低冰点、延缓冻存过程, 同时提高细胞内离子浓度,
减少细胞内冰晶的形成,从而减少细胞损伤。深低温时二甲基亚砜的细胞毒性受到抑制。复苏时动作要快，尽快洗掉二甲基亚砜，否则会造成对细胞严重的毒性。二
甲基亚砜（DMSO）是目前最好的细胞冻存保护剂，但也是一种以细胞毒性很大的化学试验剂。研究结果表明，培养液中DMSO浓度为10%时，细胞生长抑制
率近100%；1‰浓度时抑制率为35%，即使是0.04‰的浓度，DMSO对细胞的生长也有不利的影响。
1. 净化工作台
2. 离心机
3. 恒温水浴箱
4. 冰箱（4℃、-20℃、-70℃）
5. 倒置相差显微镜
6. 培养箱
7. 液氮冰箱
（二）玻璃器皿
1. 吸管（弯头、直头）
2. 培养瓶
3. 玻璃瓶（250ml、100ml）
4. 废液缸
（三）塑料器皿
1. 吸头
2. 枪头
3. 胶塞
4. 移液管（10ml）
5. 15ml离心管
6. 冻存管（1～2ml）
（四）其他物品
1. 微量加样枪
2. 红血球计数板
3. 记号笔
4. 医用橡皮膏
5. 移液枪
（五）试剂
1. D-Hanks液
2. 小牛血清
3. 培养液
4. 双抗（青霉素、链霉素）
5. 胰蛋白酶（0.08%）
6. 1NHCl
7. 7.4%NaHCO3
8. DMSO（分析纯）或无色新鲜甘油 （一）细胞冻存
1. 配制含10%DMSO或甘油、10～20%小牛血清的冻存培养液；
2. 取对数生长期的细胞，用胰蛋白酶把单层生长的细胞消化下来，悬浮生长的细胞则直接将细胞移至15ml离心管中；
3. 离心1000rpm，5min；
4. 去除胰蛋白酶及旧的培养液，加入适量配制好的冻存培养液，用吸管轻轻吹打使细胞均匀，计数，调节冻存液中细胞的最终密度为5×106/ml～1×107/ml；
5. 将细胞分装入冻存管中，每管1～1.5 ml；
6. 在冻存管上标明细胞的名称，冻存时间及操作者；
7. 冻存：标准的冻存程序为降温速率-1～-2℃/ min；当温度达-25℃以下时，可增至-5℃～-10℃/min；到-100℃时，则可迅速浸入液氮中。也可将装有细胞的冻存管放入-20℃冰箱2h ，然后放入-70℃冰箱中过夜，取出冻存管，移入液氮容器内。
（二） 细胞复苏
1. 从液氮容器中取出冻存管，直接浸入37℃温水中，并不时摇动令其尽快融化。
2. 从37℃水浴中取出冻存管，打开盖子，用吸管吸出细胞悬液，加到离心管并滴加10倍以上培养液，混匀；
3. 离心， 1000rpm，5min；
4. 弃去上清液，加入含10%小牛血清培养液重悬细胞，计数，调整细胞密度，接种培养瓶，37℃培养箱静置培养；
5. 次日更换一次培养液，继续培养。 1．从增殖期到形成致密的单层细胞以前的培养细胞都可以用于冻存，但最好为对数生长期细胞。在冻存前一天最好换一次培养液；
2．将冻存管放入液氮容器或从中取出时，要做好防护工作，以免冻伤；
3．冻存和复苏最好用新配制的培养液。向左转|向右转
