盐酸乙醇超快速分化液

Acid Alcohol Superfast Differentiation Solution

A743391-100ml

100ml

20X

室温

常规运输

室温保存，至少一年有效。

阿拉丁生产的盐酸乙醇超快速分化液是一种用于苏木素染色、苏木素伊红染色(HE染色)或Masson染色等涉及苏木素染色时，非常快速地降低细胞浆等背景性的苏木素染色并使细胞核苏木素染色更清晰的溶液。阿拉丁生产的盐酸乙醇超快速分化液，分化时间非常快，仅需2-5秒。如果希望可以精细地控制分化时间，可以考虑选购盐酸乙醇慢速分化液或盐酸乙醇快速分化液。使用本产品可获得更好的苏木素染色效果。本产品的基本原理是在酸性条件下，可以有效洗去细胞核以外的苏木素染色，从而降低背景；同时，也可以使细胞核着色过深的区域染色适当减弱，使染色更清晰。苏木素染色(hematoxylin staining or haematoxylin staining)，也被称作苏木精染色，是最常用的组织和细胞的染色方法之一。无色的苏木素(hematoxylin)氧化后形成有醌环结构(quinoid ring)的氧化苏木素(hematein or haematein)，从而可以和三价的铝离子、铁离子等形成有颜色的带正电荷的复合物(如hematein-Al3+ complexes)。氧化苏木素(也称氧化苏木精)和铝离子等形成有色的复合物的过程也被称为Dye Lake Formation。细胞核内基因组DNA的双螺旋结构中，双链上的磷酸基团向外，带负电荷，呈酸性，很容易与带正电荷的氧化苏木素复合物结合，从而形成细胞核染色。苏木素染色液有多种不同的配制方法，不同的方法可以把细胞核染色成不同深浅的蓝色或蓝紫色。苏木素是优良的细胞核染色剂', 但由于细胞质中RNA和线粒体DNA的存在，也会导致细胞质中出现非预期的染色。因此苏木素染色后经常会出现细胞质有较高背景或细胞核染色过深的情况。盐酸乙醇分化液的作用主要通过溶液中的pH值等以弱化氧化苏木素和RNA或DNA形成有色的复合物从而使细胞质中几乎没有苏木素染色，而细胞核的苏木素染色比较适中。分化过程中，细胞核中的苏木素染色在酸性条件下呈现红色，当细胞核的颜色达到适宜程度时，立刻用自来水洗涤以终止分化，此时细胞核中苏木素染色在近中性的条件下而转变为蓝色，称为返蓝或发生蓝化作用。分化的适宜程度是指经水洗返蓝后用显微镜观察，细胞核膜特别是细胞核染色质的形态结构清晰，而细胞浆及其它部分基本没有背景染色，后续再用伊红染色，就能获得细胞核与细胞浆蓝红相映、鲜艳漂亮的染色结果。如果分化不充分，细胞核为深蓝色，不能辩认出核膜及染色质的形态结构，且细胞浆等仍有浅蓝色背景，经伊红染色后即成红紫色，会影响观察效果。如果分化过度，细胞核染色会呈现浅蓝色，导致仅能看出轮廓甚至模糊不清。本盐酸乙醇分化液的组分经过反复优化，分化速度非常快，通常仅需分化2-5秒左右，可以非常快速地完成分化，特别适合对于苏木素染色分化操作非常熟练的实验人员。苏木素染色后的组织切片经本分化液分化处理后，核质着色更加清晰分明。本产品用于石蜡切片苏木素染色后的分化前后同一视野效果图参考图1。图1. 本产品用于小鼠睾丸石蜡切片苏木素染色后同一视野分化前后的实拍效果图。左图为未经分化处理的效果图，右图为经本产品分化处理后的效果图。从图中可见，经本产品分化处理后，苏木素染色的细胞核呈现蓝色，核质清晰分明，背景呈现纯净的白色。本图仅作参考，不同的样品不同的检测条件，实际获得的结果可能有所差别。Scale bar, 100μm。按照每个样品使用0.2ml盐酸乙醇分化液计算，一个包装的C0165S、C0165M和C0165L分别可以分化500个、2500个和10000个样品。如果用于浸泡式的分化操作，建议在使用约10-20次并且分化效果明显减弱后更换分化液。 注意事项 ： 本产品易挥发，请注意密闭保存。为避免挥发导致的使用效果下降，开启后宜尽快用完。染色后的分化为选做步骤，但分化后核质染色更清晰，核膜及染色质的形态结构更清楚。样品数量很多时，可以使用阿拉丁生产的染色架(FJ024、FJ026、FJ030、FJ060)和染色缸，便于操作。如果订购的是C0165L盐酸乙醇超快速分化液(20X)，需要自备75%乙醇，以用于把盐酸乙醇超快速分化液(20X)稀释20倍配制成盐酸乙醇超快速分化液。不同样品和不同染色要求，所需要的分化时间会有所不同。初次使用本产品建议先摸索不同分化时间，根据镜下观察结果，确定最优的分化时间。也可以根据需求选择阿拉丁的盐酸乙醇慢速分化液或盐酸乙醇快速分化液。本产品为易燃液体，须密闭储存于阴凉、干燥、通风良好的地方，建议存放在易燃液体的专用安全柜中，并远离明火、火花、热源、热表面，禁止吸烟。本产品有腐蚀性，操作时请小心，并注意有效防护以避免直接接触人体或腐蚀其他物品。本产品仅限于专业人员的科学研究用，不得用于临床诊断或治疗，不得用于食品或药品，不得存放于普通住宅内。为了您的安全和健康，请穿实验服并戴一次性手套操作。 使用说明 ： 1.需要用户自己准备的试剂：a.固定液：阿拉丁的免疫染色固定液，或4%多聚甲醛固定液。b.苏木素染色液：阿拉丁的苏木素染色液或苏木素伊红染色试剂盒；c.如果需要脱水、透明和封片处理，还需自备二甲苯和封片剂，如阿拉丁的PVP封片液、抗荧光淬灭封片液或中性树胶等其它封片剂。d.70%乙醇，如果样品是石蜡切片或需要脱水、透明、封片处理，需自备80%乙醇、90%乙醇、无水乙醇以及二甲苯。如果使用的是C0165L盐酸乙醇超快速分化液(20X)，需要自备75%乙醇，以用于把盐酸乙醇超快速分化液(20X)稀释20倍配制成盐酸乙醇超快速分化液，例如100ml盐酸乙醇超快速分化液(20X)加入1.9L 75%乙醇，混匀后即为盐酸乙醇超快速分化液。e.蒸馏水。2.样品处理：a.对于石蜡切片：二甲苯中脱蜡5-10分钟。换用新鲜的二甲苯，再脱蜡5-10分钟。无水乙醇5分钟。90%乙醇2分钟。80%乙醇2分钟。70%乙醇2分钟。蒸馏水2分钟。b.对于冰冻切片：固定液固定10分钟以上。蒸馏水2分钟。换用新鲜的蒸馏水，再洗涤2分钟。c.对于培养细胞：固定液固定10分钟以上。蒸馏水洗涤2分钟。换用新鲜的蒸馏水，再洗涤2分钟。3.苏木素染色：对于上述处理好的样品：a.苏木素染色液染色5-10分钟(可以根据染色结果和要求适当调整染色时间)。b.浸自来水中冲洗去多余的染色液，约10分钟。c.蒸馏水再洗涤一遍(约数秒钟)。d.盐酸乙醇超快速分化液分化约2-5秒(可以根据分化结果和要求适当调整分化时间)。自来水冲洗返蓝10分钟。此时，如果需要直接观察，可以用70%乙醇洗涤2次。如需进行伊红染色，在返蓝后即可按照伊红染色液的操作步骤进行伊红染色。如需脱水、透明后封片，按常规步骤进行，70%乙醇洗涤后仍可按照后续步骤进行脱水、透明和封片处理。注：如果用于免疫组化等染色后的复染，可以参考上述步骤在其它染色完成后直接进行苏木素染色。 Aladdin's Acid Alcohol Superfast Differentiation Solution is generally used in hematoxylin staining, hematoxylin-eosin staining (HE staining) or Masson staining to reduce the staining background from cytoplasm, making the hematoxylin stain of nucleus more lucid.This product developed by aladdin enables a very fast differentiation of samples, requiring only 2-5 seconds. If a slower differentiation is desired, we recommend using the Acid Alcohol Fast Differentiation Solution or the Acid Alcohol Slow Differentiation Solution .A better hematoxylin stainng result can be obtained if the differentiation step is performed. The Acid Alcohol Differentiation Solution reduces the stain background effectively by removing the hematoxylin stain outside nucleus. Meanwhile, it also weakens the over-stained area in nucleus to improve the hematoxylin staining result of nucleus.Hematoxylin staining is one of the most commonly used methods for staining tissues and cells. The colorless hematoxylin is oxidized to form hematein with a quinoid ring structure, which can form a colored positively charged complex with trivalent aluminum ions, iron ions, etc. In the DNA double helix in nucleus, the phosphate groups on double strands are outward, negatively charged and acidic, and easily bind with the positively charged hematein complex to stain nucleus in purple. Hematoxylin Staining Solution prepared in different ways can stain cell nuclei in blue or blue-violet with different intensities.Hematoxylin is an excellent dye for staining nucleus. However, unexpected cytoplasm staining generally occurs due to the presence of RNA and mitochondrial DNA in cytoplasm, resulting in a high cytoplasm stain background or an over-staining of nucleus. The function of the Acid Alcohol Differentiation Solution is to weaken the colored complex of oxidized hematoxylin and RNA or DNA through the pH value of the solution, so that there is almost no hematoxylin staining in the cytoplasm while the hematoxylin staining of nucleus is relatively moderate. During the differentiation process, the hematoxylin stain in nucleus appears red under acidic conditions. When nucleus staining reaches an appropriate level, the differentiation is stopped immediately by washing samples with tap water and the hematoxylin stain in nucleus returns blue. The appropriate differentiation means that nucleus membrane and chromatin in nucleus especially have clear morphology while the cytoplasm and other parts have basically no background staining. Cytoplasm can be stained in red in contrast by the subsequent eosin staining. Insufficient or over differentiation should be avoided by adjusting the differentiation time properly.The formula of this product has been optimized to have a superfast differentiation. It is especially suitable for researchers who are very skilled in hematoxylin staining and differentiation procedures. The hematoxylin staining result is improved significantly by the addition of differentiation step. Please see Figure 1 for the hematoxylin staining result before or after differentiation by the Acid Alcohol Superfast Differentiation Solution .Figure 1. Paraffin sections of mouse testis stained with hematoxylin and observed before differentiation (Left) or after differentiation (Right) by the Acid Alcohol Superfast Differentiation Solution . Sample after differentiation (Right) has reduced background from cytoplasm, and clearer nuclei, compared to the sample before differentiation (Left). Scale bar, 100μm. This figure is for reference only, the actual results may vary due to different experimental conditions. When 0.2ml of Differentiation Solution is used for each sample, C0165S, C0165M and C0165L products are sufficient for the differentiation of 500, 2500 and 10,000 samples, respectively. When using the immersion method for differentiation, the Differentiation Solution can be resued for 10-20 times until its performance decreases significantly. Precautions ： This product is volatile. Please keep the vial tightly closed after use. The reagent should be used up as soon as possible after opening.The differentiation step after staining is optional, but if the differentiation step is performed, the nucleus staining result will be better with clearer morphological structure of nuclear membrane and chromatin.When there is a large number of samples, staining racks (FJ024, FJ026, FJ030, FJ060) or tanks produced by can be used for easier operation.The C0165L product, the Acid Alcohol Superfast Differentiation Solution (20X), needs to be diluted to 1X with 75% ethanol prior to use.The duration period of differentiation should be optimized for different samples based on your experimental requirements. For first-time users of this product, it is recommended to perform a time course experiment to determine the best differentiation time for your samples. The Acid Alcohol Fast Differentiation Solution and the Acid Alcohol Slow Differentiation Solution are also available for different experimental requirement.This product is volatile and flammable, and must be stored hermetically in a cool, dry and well-ventilated safety cabinet away from open fire, spark, heat source, and hot surface. Smoking is prohibited.This product is corrosive. Please be careful when handling the reagent and reactions, and take effective measures to avoid direct contact or corrosion of other objects.This product is for R&D only. Not for drug, household, or other uses.For your safety and health, please wear a lab coat and disposable gloves during the operation.Materials Required But Not Supplieda. Fix Solution: Immunol Staining Fix Solution , or 4% Paraformaldehyde Fix Solution .b. Hematoxylin Staining Solution: Hematoxylin Staining Solution or Hematoxylin and Eosin Staining Kit .c. If dehydration, clearing and mounting are required, xylene and mounting medium can be purchase from , such as Polyvinylpyrrolidone Mounting Medium , Antifade Mounting Medium , neutral gum or other mounting media.d. Different concentrations of ethanol: Absolute ethanol, 90% ethanol, 80% ethanol, 75% ethanol.e. Distilled water. Instructions for Use ： 1. Sample preparation: a. For paraffin sectionsDewax paraffin sections in xylene for 5-10 minutes;Replace with fresh xylene and dewax for another 5-10 minutes;Anhydrous ethanol for 5 minutes;90% ethanol for 2 minutes;80% ethanol for 2 minutes;70% ethanol for 2 minutes;Distilled water for 2 minutes;Proceed to step 2.b. For frozen sectionsFix frozen sections in Fix Solution for more than 10 minutes;Wash twice with distilled water for 2 minutes each;Proceed to step 2.c. For cultured cellsFix cells in Fix Solution for more than 10 minutes;Wash cells twice with distilled water for 2 minutes each;Proceed to step 2.2. Hematoxylin staining:a. Stain samples with Hematoxylin Staining Solution for 5-10 minutes (Time can be adjusted appropriately).b. Rinse samples with tap water to remove the extra staining solution, and immerse samples in tap water for 10 minutes.c. Wash with distilled water once for a few seconds.d. Perform sample differentiation with the Acid Alcohol Superfast Differentiation Solution for 10 seconds, and then rinse with tap water for 10 minutes. The differentiation time can be adjusted appropriately.e. Wash samples with 70% ethanol twice, examine cells directly or proceed to eosin staining or sample mounting after dehydration and clearing. Note: For counterstaining of sample that has been subdued to immunohistochemistry staining, the hematoxylin staining can be followed directly after completing immunohistochemistry staining.