胭脂红又称胭脂虫红、卡红、洋红等，英文名称为Carmine，水溶性合成色素，鲜艳的黄光红色，单色品种。可用于食品、饮料、药品、化妆品、饲料、烟草、玩具、食品包装材料等的着色。在水中呈深红色；在酸性溶液中呈黄色至紫色。溶于水、甘油、浓硫酸，微溶于乙醚、乙醇，几乎不溶于石油醚、苯、氯仿、油脂。
天然胭脂红来源于热带的一种雌性胭脂虫干燥后，磨成粉末，提取出胭脂红，再用明矾处理，除去其中杂质，制成胭脂红。单纯的胭脂红不能染色，要经酸性或碱性溶液溶解后才能染色。常用的酸性溶液有冰醋酸或苦味酸，碱性溶液有氨水、硼砂等。胭脂红是细胞核的优良染料，染色的标本不易褪色。可用作切片或组织块染，尤其适宜于小型材料的整体染色。
盐酸卡红染色液 (2%)又称盐酸洋红染色液或盐酸胭脂红染色液，由卡红、盐酸、酒精、去离子水等组成，pH呈酸性。盐酸卡红染色液主要用于除大型标本外的其他寄生虫样本的染色观察，也可用于植物染色体的研究。该试剂仅用于科研领域，不适用于临床诊断或其他用途。
自备材料：
1、载玻片、盖玻片、恒温箱、光学显微镜
2、卡诺固定液、45%醋酸、1%盐酸乙醇分化液、梯度乙醇、二甲苯、中性树胶等
操作步骤(仅供参考)：
(一)用于植物染色体染色：
1、样本固定：可取植物根尖、幼叶、幼芽、茎尖、幼嫩的子房和花药等作为样本。花药不需任何前处理。用于有丝分裂中期染色体核型分析的材料，需经常规预处理，子房的处理时间比根尖长1~2小时，或者使用的秋水仙素和8-羟基喹啉溶液的浓度比根尖的使用浓度高1倍，处理时间不变。预处理后的材料用卡诺固定液固定24小时以上。
2、洗涤：用70%乙醇洗涤材料2~3次，每次1小时以上。
3、染色：滤纸快速吸干材料表面，放入盐酸洋红中，染液体积一般为材料的3倍以上，染色时间依材料而定。花药染色时间为12~24小时，其他材料如子房、幼叶等为2~6天。如需缩短染色时间，可放入60°C温箱中染色1~3天。
4、染色后洗涤：染好的材料表面看是紫红色，再用70%乙醇洗涤材料2~3次，每次10分钟，洗涤好可保存于70%乙醇中。染液可重复使用数次。
5、软化和分色：用45%醋酸在60°C条件下软化10~30分钟，软化时间需依材料而定。软化后的材料可用新的45%醋酸在酒精灯上分色5~20秒。
6、压片：取出软化和分色后的材料放于载玻片上，滴加一滴45%醋酸，常规压片，镜检。
7、永久制片：冰冻揭片、气干或在温箱中低温烘干，中性树胶封片。
(二)用于寄生虫样本的染色：
1、样本洗涤：可取除大型标本外的其他寄生虫作为样本。自保存液中取出寄生虫，用70%乙醇洗涤材料2~3次，每次1小时以上。
2、染色：滤纸快速吸干虫体表面，放入盐酸洋红中，染液体积一般为材料的3倍以上，染色时间依材料而定，一般为6~12小时，如需缩短染色时间，可放入60°C温箱中。
3、分色：取出虫体，放入1%盐酸乙醇中快速分色10~30秒，再用70%乙醇继续分色2~3次，每次10分钟。染液可重复使用数次。
4、脱水透明：依次使用80%乙醇2~12小时、95%乙醇2~12小时(时间依材料大小而定)，再依次入新的95%乙醇1小时、无水乙醇1小时(两次)、二甲苯10分钟(两次)，中性树胶封片观察。
注意事项：
1、进行有丝分裂中期核型分析材料时，幼叶、茎尖、花药壁、子房均可，但子房最佳。
2、材料经固定后，一定要用70%乙醇充分洗尽固定液。
3、该试剂含有高浓度乙醇，有强挥发性，且染色时间较长，宜用玻片盒或带盖染色缸密封染色。
4、注意密闭保存，否则染色效率会下降。
5、为了您的安全和健康，请穿实验服并戴一次性手套操作。
Carmine, also known as cochineal red, carmine red, or magenta, is a water-soluble synthetic pigment. It presents a bright yellowish red color and belongs to the monochromatic pigment category. It can be used for coloring food, beverages, pharmaceuticals, cosmetics, feed, tobacco, toys, food packaging materials, and other products. In water, it appears as a deep red color; in acidic solutions, it turns from yellow to purple. It is soluble in water, glycerol, and concentrated sulfuric acid, slightly soluble in ether and ethanol, and almost insoluble in petroleum ether, benzene, chloroform, and oils.
Natural carmine is derived from dried female cochineal insects native to tropical regions. The insects are ground into powder, from which carmine is extracted, and then treated with alum to remove impurities, yielding the final carmine product. Pure carmine cannot act as a stain on its own; it must first be dissolved in an acidic or alkaline solution to become functional. Commonly used acidic solutions include glacial acetic acid or picric acid, while common alkaline solutions include ammonia water and borax. Carmine is an excellent nuclear stain, and specimens stained with it are resistant to fading. It can be used for staining sections or tissue blocks, and is particularly suitable for whole-mount staining of small specimens.
Carmine Hydrochloride Staining Solution (2%), also called carmine hydrochloride stain or hydrochloric acid carmine stain, is composed of carmine, hydrochloric acid, alcohol, and deionized water, with an acidic pH. This staining solution is mainly used for the staining and observation of parasitic samples excluding large specimens, and can also be applied to the study of plant chromosomes. This reagent is intended for research use only and is not suitable for clinical diagnosis or any other purposes.
Materials to Be Prepared by User:
1. Glass slides, cover slips, incubator, optical microscope
2. Carnoy's fixative, 45% acetic acid, 1% hydrochloric acid-ethanol differentiation solution, graded ethanol series, xylene, neutral balsam, etc.
Operating Procedures (For Reference Only):
(I) For Plant Chromosome Staining
1. Sample Fixation: Plant root tips, young leaves, buds, shoot tips, tender ovaries, and anthers can be used as samples. Anthers require no pre-treatment. For materials intended for karyotype analysis of metaphase chromosomes during mitosis, routine pre-treatment is necessary. The treatment duration for ovaries should be 1–2 hours longer than that for root tips; alternatively, the concentrations of colchicine and 8-hydroxyquinoline solutions used can be doubled while keeping the treatment duration unchanged. Pre-treated materials should be fixed in Carnoy's fixative for more than 24 hours.
2. Washing: Rinse the materials 2–3 times with 70% ethanol, with each rinse lasting over 1 hour.
3. Staining: Blot the surface of the materials dry quickly with filter paper and immerse them in carmine hydrochloride. The volume of the staining solution should generally be more than 3 times that of the materials, and the staining duration depends on the material type. The staining time for anthers is 12–24 hours, while for other materials such as ovaries and young leaves, it is 2–6 days. To shorten the staining time, the samples can be placed in an incubator at 60°C for 1–3 days of staining.
4. Post-staining Washing: Well-stained materials appear purplish-red on the surface. Rinse the stained materials 2–3 times with 70% ethanol, 10 minutes each time. The washed materials can be preserved in 70% ethanol. The staining solution can be reused several times.
5. Softening and Differentiation: Soften the materials in 45% acetic acid at 60°C for 10–30 minutes, with the exact duration depending on the material. After softening, differentiate the materials in fresh 45% acetic acid over an alcohol lamp for 5–20 seconds.
6. Squashing: Place the softened and differentiated materials on a glass slide, add one drop of 45% acetic acid, perform squashing following standard procedures, and examine under a microscope.
7. Permanent Slide Preparation: Carry out freeze peeling, air drying, or low-temperature drying in an incubator, then mount the slide with neutral balsam for permanent preservation.
(II) For Parasite Sample Staining
1. Sample Washing: Parasite samples excluding large specimens can be used. Remove the parasites from the preservation solution and rinse them 2–3 times with 70% ethanol, with each rinse lasting over 1 hour.
2. Staining: Blot the surface of the parasites dry quickly with filter paper and immerse them in carmine hydrochloride. The volume of the staining solution should generally be more than 3 times that of the samples. The staining duration depends on the sample type, usually 6–12 hours. To shorten the staining time, the samples can be incubated at 60°C.
3. Differentiation: Take out the parasites, differentiate them rapidly in 1% hydrochloric acid-ethanol for 10–30 seconds, then continue differentiation in 70% ethanol for 2–3 times, 10 minutes each time. The staining solution can be reused several times.
4. Dehydration and Clearing: Dehydrate the samples sequentially in 80% ethanol for 2–12 hours and 95% ethanol for 2–12 hours (duration varies with sample size). Then transfer the samples successively to fresh 95% ethanol for 1 hour, absolute ethanol twice for 1 hour each time, and xylene twice for 10 minutes each time. Finally, mount the slide with neutral balsam for microscopic observation.
Precautions:
1. For materials intended for metaphase mitosis karyotype analysis, young leaves, shoot tips, anther walls, and ovaries are all applicable, with ovaries being the optimal choice.
2. After the materials are fixed, it is essential to thoroughly rinse off the fixative with 70% ethanol.
3. This reagent contains a high concentration of ethanol, which is highly volatile. In addition, the staining process requires a relatively long time, so it is advisable to perform sealed staining using slide boxes or covered staining jars.
4. Store the reagent in an airtight container; otherwise, the staining efficiency will decrease.
5. For your safety and health, wear a lab coat and disposable gloves during the operation.
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