HE Staining
HE染色资料介绍
普通染色又称常规染色或HE（Haematoxylin and eosin）染色。
它是病理技术中最常用的一种方法，通过它可以做出病理诊断和发现寻求别的辅助方法，以达到准确，完整的病理诊断。
一、染色目的病理学的所有切片，都必须通过一种以上的染料，通过各种不同的方法，将切片中各种不同的物质，在不同染液的作用下，将其显示出来，使之在光学显微镜下，能够完全的观看各种结构。
例如，HE染色，好质量的切片可以清晰地显示出许多不同的结构，细胞核着蓝黑色，细胞浆着粉红色，软骨着蓝色等。
清晰的结构为诊断提供可靠的依据，因此，染色技术也是病理技术中的重要组成部分，必须不断地总结，方能提高。
如果染色不好，切片染色一团糟，红蓝不分，结构不清，层次不明，影响了镜下的观察，直接影响了临床诊断，染色结果的好坏直接关系到诊断的准确性。
二、染色的作用1．化学作用：所有的染色液中，可把它们分为两种类型，一种为酸性，另一种为碱性。
酸性染料中有染色作用的为阴离子，碱性染料中有染色作用的为阳离子，每个细胞也存在着两种物质，细胞核含的是酸性物质，细胞浆含的是碱性物质。
在染色时，细胞核中的酸性物质与苏木素染液中的阳离子发生作用，细胞浆中的碱性物质与伊红染液中的阴离子发生作用，由于反应的部位不同，结果着色有异。
2．物理作用：在染色过程中，染液中的色素微粒子浸入到被染组织的粒子间隙内，此时，因受分子的引力作用，色素微粒子被吸附而着色。
由于各种组织有不同的吸附能力和不同的吸附程度，因此就可显出来不同的颜色来。
一般来说，染色的学说还有许多，但说服力强的仅有上述两种。
但不管怎么样解释，实际上完成的每一种染色，都与上述两种学说分不开，它们的作用是相辅相成，同时存在的。
三、染色方法及步骤1．人工苏木素-伊红染色法（HE法）(1)切片浸入二甲苯中5-10min；(2)切片浸入二甲苯中5-10min；(3)100%酒精1min。
(4)100%酒精1min。
(5)95%酒精1min。
不同温度对HE染色效果的影响
醇 １００ ｍＬ、硫酸铝钾 １５０ ｇ、蒸馏水 ２ ０００ ｍＬ、碘酸钠 １．２ ｇ、冰 醋酸 １２０ ｍＬ、甘油 ９００ ｍＥ）、０．５％ ～１．０％伊红染 色液 （９５％ 乙醇配制 ）等。 １．１．３ 试验仪器 本试验所 用的仪器主要有 精密 电子 天平 （Ａｔｅ）、电热鼓风干燥箱 （１０１—５型）、包埋机（徕卡 ＹＢ一６ＬＦ）、 轮转式切片机（徕卡 ＲＭ２０１６）、摊片机（徕卡 １－１１１２１０）等。
１ 材料与方法
１．１ 材 料 与 仪 器 １．１．１ 材料 草鱼 的肾、肝、肠组织切片。 １．１．２ 试 剂配制 常 用于 组织学 切片 制作 的试剂 有 Ｂｏｕｉｎ 氏液（苦味酸 ７５ ｍＬ、４０％ 甲醛 ２５ ｍＬ、冰醋酸 ５ ｍＬ）、粘合剂 （鸡蛋清 ）、清洗液 （重 铬酸钾 ８Ｏ ｇ、水 １ ０００ ｍＬ、粗制 浓硫 酸 １２０ ｍＬ）、乙醇 （分别 配制 了 ７０％、８０％、９０％ 、９５％ 、１００％ ５ 种浓度 ）、１／２二 甲苯 （乙醇和二 甲苯按 １：ｌ比例配制）、氨水 （１％）、酸水 （１％ ）、０．２％苏 木精染色液 （苏木 精 ６ ｇ、无水 乙
·－—
—
１６８－ ——
江苏农业科学 ２０１７年第 ４５卷第 ２２期
王庆容 ，徐 晓舒 ，夏 浪 ，等．不同温度 对 ＨＥ染色效果的影响［Ｊ］．江苏农业科学，２０１７，４５（２２）：１６８—１７０ ｄｏｉ：１０．１５８８９／ｊ．ｉｓａｎ．１００２—１３０２．２０１７．２２．０４６
不 同温度对 ＨＥ染 色效果 的影 响
收稿 日期 ：２０１６—０３—１７ 基金项 目：贵州省科技 厅农业 攻关项 目（编号 ：黔科合 ＮＺ［２０１３］３０２７
号 ）；贵州省教育 厅“１２５计划”重大科技专项 （编号 ：黔教合重大专 项字 ［２０１３］０２５号 ）；贵 州省科技 厅联 合基金 （编号 ：黔 科合 Ｊ字 ＬＫＺＳ［２０１２］１７号 ）。 作者简介 ：王庆容（１９７２一 ），女 ，贵州务川人 ，硕 士，教授 ，主要从事动 物分子生物学及疾病防治研 究。Ｅ—ｍａｉｌ：６１０１９４９７７＠ｑｑ．ＣＯＩＴＩ。
石蜡切片制作和HE染色实验报告
石蜡切片制作和HE染色实验报告英文回答：Introduction:Paraffin sectioning and H&E staining are essential techniques in histopathology for examining tissue samples under a microscope. Paraffin sectioning involves embedding tissues in paraffin wax, cutting thin sections, and mounting them on glass slides. H&E staining is a common staining method that uses hematoxylin and eosin to differentiate cell components.Materials and Methods:Tissue samples。
Paraffin wax。
Microtome。
Glass slides。
Hematoxylin。
Eosin。
Ethanol。
Xylene。
Procedure:Paraffin Sectioning:1. Fix the tissue samples in formalin and process them through a series of graded ethanol solutions.2. Embed the tissues in paraffin wax and allow them to solidify.3. Use a microtome to cut thin sections (5-10 microns)from the paraffin block.4. Mount the sections on glass slides and allow them to dry.H&E Staining:1. Deparaffinize the sections by passing them through xylene and ethanol solutions.2. Stain the sections with hematoxylin and rinse with water.3. Differentiate the hematoxylin staining with hydrochloric acid and rinse with water.4. Counterstain the sections with eosin and rinse with water.5. Dehydrate the sections by passing them through graded ethanol solutions and clear them in xylene.6. Mount the sections with a coverslip and examine them under a microscope.Interpretation of Results:Hematoxylin stains the nuclei blue, while eosin stains the cytoplasm pink.The stained sections allow for the examination of tissue morphology, identification of cell types, and detection of pathological changes.Conclusion:Paraffin sectioning and H&E staining are fundamental techniques in histopathology that provide valuable information for the diagnosis and study of diseases.中文回答：石蜡切片制作和HE染色实验报告。
HE染色
结果的判断
实验结果
细胞核被苏木精染成鲜明的蓝色，软骨基质、钙盐颗粒呈深蓝色，粘液呈灰蓝色。细胞浆被伊红染成深浅不 同的粉红色至桃红色，胞浆内嗜酸性颗粒呈反光强的鲜红色。胶原纤维呈淡粉红色，弹力纤维呈亮粉红色，红血 球呈橘红色，蛋白性液体呈粉红色。
着色情况与组织或细胞的种类有关，也随其生活周期及病理变化而改变。例如，细胞在新生时期胞浆对伊红 着色较淡或轻度嗜碱，当其衰老时或发生退行性变则呈现嗜伊红浓染。胶原纤维在老化和出现透明变性时，伊红 着色由浅变深。
H-E染色评定标准：
（1）切片完整，厚度4-6微米，厚薄均匀，无皱褶无刀痕；
（2）染色核浆分明，红蓝适度，透明洁净，封裱美观。
谢谢观看
由于组织或细胞的不同成分对苏木精的亲和力不同及染色性质不一样。经苏木精染色后，细胞核及钙盐粘液 等呈蓝色，可用盐酸酒精分化和弱碱性溶液显蓝，如处理适宜，可使细胞核着清楚的深蓝色，胞浆等其它成分脱 色。再利用胞浆染料伊红染胞浆，使胞浆的各种不同成分又呈现出深浅不同的粉红色。故各种组织或细胞成分与
试剂与仪器
适用范围
组织学、胚胎学、生物学、病理学教学与科研。
原理
易于被碱性或酸性染料着色的性质称为嗜碱性( basophilia )和嗜酸性( acidophilia )；而对碱性染料和 酸性染料亲和力都比较弱的现象称为中性（neutrophilia)。
构成组织内蛋白质的氨基酸的种类很多，它们有不同的等电点。在普通染色法中，染色液的酸碱度为pH6左 右，细胞内的酸性物质如细胞核的染色质、腺细胞和神经细胞内的粗面内质及透明软骨基质等均被碱性染料染色， 这些物质称为嗜碱性。而细胞质中的其它蛋白质如红细胞中的血红蛋白、嗜酸粒细胞的颗粒及胶原纤维和肌纤维 等被酸性染料染色，这些物质称为嗜酸型。如果改变染色液的酸碱度，pH值升高时，则原来被酸性染料染色的物 质可变为嗜碱性；pH值降低时，原来被碱性染料染色的物质则可变为嗜酸性。所以说染色液的pH值可以影响染色 的反应。
he染色原理和流程
he染色原理和流程English.Hematoxylin and Eosin (H&E) Staining Principle and Procedure.Hematoxylin and eosin (H&E) staining is a widely used histological staining technique that employs hematoxylin and eosin dyes to differentially stain the nuclei and cytoplasmic components of cells, respectively. This staining method provides a clear visualization of cellular structures, enabling the identification and characterization of tissues and cell types in tissue sections.Principle of H&E Staining.H&E staining relies on the interaction between the basic dye hematoxylin and acidic tissue components, primarily DNA and RNA. Hematoxylin stains these nucleicacids blue or purple, highlighting the nuclei of cells. In contrast, the acidic dye eosin binds to basic structures such as proteins, staining the cytoplasm and extracellular matrix in shades of pink or red. This differential staining allows for the clear distinction between cellular compartments and provides insights into the cellular composition and morphology.Procedure for H&E Staining.H&E staining involves a series of sequential steps:1. Tissue Preparation: Tissue samples are processed to create thin sections suitable for staining. This involves various steps such as fixation, embedding, sectioning, and mounting the sections on slides.2. Deparaffinization and Rehydration: If the tissue sections are paraffin-embedded, they undergo deparaffinization to remove the paraffin wax. Subsequently, the sections are rehydrated by passing them through a series of graded alcohol baths.3. Hematoxylin Staining: The tissue sections are immersed in hematoxylin solution, which stains the nuclei blue or purple. The staining time can be adjusted to achieve the desired intensity.4. Differentiation and Bluing: The hematoxylin staining is differentiated using acid alcohol or hydrochloric acid to remove excess dye and enhance the nuclear staining. Subsequently, the sections are treated with ammonia or sodium bicarbonate solution to "blue" the hematoxylin, resulting in a sharp nuclear staining.5. Eosin Counterstaining: The tissue sections are rinsed and counterstained with eosin solution, which imparts a red or pink color to the cytoplasm and extracellular components.6. Dehydration and Clearing: The stained sections are dehydrated by passing them through a series of graded alcohol baths. They are then cleared in a solvent such as xylene or toluene.7. Mounting: The cleared sections are mounted on slides using a resinous mounting medium for permanent preservation and visualization under a microscope.Applications of H&E Staining.H&E staining is a versatile technique with numerous applications in histology, including:Routine examination of tissues for identification of normal and abnormal structures.Diagnosis of various diseases, such as cancer, inflammation, and infections.Evaluation of tissue architecture and cell morphology.Research in cell biology, pathology, and developmental biology.中文回答：苏木精-伊红 (H&E) 染色原理和流程。
名词解释-问答
1.Introduction●HE staining: a staining method with hematoxylin and eosin. Basophilic substances incells will be stained by hematoxylin and show a blue color, while acidophilic substances will be stained by eosin and show a red color.2.Epithelium●Endothelium: simple squamous epithelium lining of the inner surfaces of circulatorysystem(heart, vessels, lymph vessels ) that facilitate fluid flow and the exchange of substances●Mesothelium:simple squamous epithelium lining of the outer surfaces of bodycavities(pericardium, pleura, peritoneum) that are moist, which help avoid mechanical friction between an organ and another●Brush/striated border:many microvilli get together and can be seen in LM●Junction complex: 2 or more type of cell junctions(tight junction, gap junction,intermediate junction and desmosome) at the same lateral surface of cell●Plasma membrane infolding: the cell membrane of basal surface invaginate(陷入) intocell body in order to enlarge the surface of cell; it’s rich in mitochondrion●Compare cilium and microvilli:Both are finger-like processes on the free surface of epithelium and they can enlarge the surface of cells. Microvilli can help the absorption of cells, while cilium can make fluid flow in one direction by their rapid back and forth movement3.connective tissue●fibrocyte: fibroblast in rest form, can be transmitted into fibroblast●molecular sieve: the structure of proteoglycan sub-units are rich in mesh(网眼) ,whichfiltrate molecules of different sizes and therefore act as a barrier to prevent the spread of bacteria and other micro-organisms●Describe the function and structure of plasma cell and macrophage:Plasma cell: ovoid cell with basophilic cytoplasm and eccentric located nucleus; play a role in the production of antibody after the recognition of antigen, therefore involved in immune reactionMacrophage: irregular shaped with short, blunt processes and a kidney-shaped nucleus;rich in lysosome; phagocytosis, secretion and as antigen-presenting cell4.bone and cartilage●distribution of 3 types of cartilage:hyaline cartilage: temporary skeleton of embryo and the main type of cartilage for adults(transparent, white or faded blue)elastic cartilage: auricle耳廓,pharynx咽喉,epiglottis会厌(not transparent, yellow)fibrous cartilage: intervertebral disc, pubic synthesis,meniscus, articular discs,etc. (milk white)●maturation process of osteocyte:mesenchymal stem cell----osteoprogenitor cell----osteo b last----osteocyte/bone lining cell(also: osteo c last)●osteoid类骨质: Uncalcified ECM of osseous tissue and it is secreted by osteoblast. It willchange to bone matrix once being calcified. In HE staining, it appears a thin paler line atthe surface of osseous tissue and below the osteoblasts.●osteons(Haversian systems)骨单位骨板: the major structure of long bone locatingbetween outer and inner circumferential laminae. They are concentric laminae surrounding central canal containing blood vessels, nerves and loose connective tissue.5.blood and haemopoiesis●neutrophil: neutrophilic granulocyte, the most numerous leukocyte in human blood;segmented nucleus with2-5 lobes; have azurophilic granules and specific granules; playa role in acute inflammation and defend body against the micro-organisms●who take part in allergy: eosinophil has enzymes that can dissolve histamine andleukotriene to inhibit allergy; basophil has leukotriene in its cytoplasm●reticulocyte: immature erythrocytes with ribosomes left in their cytoplasm, about onepercent of the total number of RBC;6.muscle tissue●Sarcomere: segment of the myofibril between two Z lines, consists of two halves of Iband and one A band. It’s the structural and functional unit of myofibril.●Sarcoplasmic reticulum: specialized SER, longitudinal aligned, enclose the myofibril,store and release calcium●Sarcoplasmic reticulum: specialized SER of muscle fiber that are longitudinal aligned andthey enclose the myofibril. Their function is to store and release calcium, especially during muscle contraction.●Terminal cisternae: expanded sarcoplasmic reticulum at the boundaries of A and I bands●T tubule: sarcolemma enfolds and penetrates into sarcoplasm●Triad: T tubule + two closely associated terminal cisternae; there’re two sets of triads inone sarcomere at the boundaries of A and I band; there function is to transmit impulses from sarcolemma into sarcoplasmic reticulum.●Intercalateddisk: junctional complexbetween cardiac muscle fibers, cross striationacross the cell under LM; at the level of Z line; it’s passage for information that contributes to the synchronization同步化of cardiac muscle fiber impulses.7.Nerve tissue●Compare dendrite with axon:There’s only one axon in most neurons but more dendrites; Dendrites are short and divided while axon is long and cylindrical. Dendrites are principal signal receptors and processing sites that●Blood-brain barrier: expanded foot plates of astrocytes that cover capillary endothelialcells, forming glia limitans that regulate the diffusion of substances between blood and the brain, which protect neurons from harmful substances in blood.●Ranvier node:a naked part of axon between adjacent Schwann cells that is not wrappedby neurolemma and mylinated sheath●Nissel body: strong basophilic clumps under LM; RER and free ribosomes that activelyproducing cytoskeletal protein, enzymes for neurotransmitters and peptide neuromodulators.。
石蜡切片制作和HE染色实验报告
石蜡切片制作和HE染色实验报告英文回答：Introduction。
Paraffin sectioning and HE staining are fundamental techniques used in histology to prepare tissue samples for microscopic examination. Paraffin sectioning involves embedding the tissue in paraffin wax, which allows for thin sections to be cut using a microtome. HE staining, also known as hematoxylin and eosin staining, is a common staining method used to differentiate between differentcell types and structures in the tissue. Together, these techniques provide a detailed view of the tissue's morphology and architecture.Materials and Methods。
Paraffin Sectioning。
1. Fix the tissue in formalin for 24 hours.2. Dehydrate the tissue by passing it through a seriesof graded alcohols (70%, 95%, 100%).3. Clear the tissue by passing it through xylene.4. Embed the tissue in paraffin wax.5. Cut thin sections (5-10 µm) using a microtome.HE Staining。
01-组织学绪论
（四） 组织化学和细胞化学技术
组织化学（histochemistry）和细胞化学（cytochemistry） 组织化学（histochemistry）和细胞化学（cytochemistry）技 术是应用化学反应原理检测组织和细胞的化学成分并进行 定位和定量的技术。组织细胞中的糖类、脂类、蛋白质、 定位和定量的技术。组织细胞中的糖类、脂类、蛋白质、 核酸、酶等均可与相应试剂反应， 核酸、酶等均可与相应试剂反应，最后形成有色反应终产 物或电子致密物，应用光镜或电镜进行观察。 物或电子致密物，应用光镜或电镜进行观察。 糖类物质常用过碘酸-Schiff反应(PAS反应)显示（ 糖类物质常用过碘酸-Schiff反应(PAS反应)显示（图1-4）。 脂类常用苏丹染料、油红O 尼罗蓝等脂溶性染料染色。 脂类常用苏丹染料、油红O、尼罗蓝等脂溶性染料染色。
（六）原位杂交
原位杂交是一种在组织细胞原位进行的核酸分子杂交技术， 原位杂交是一种在组织细胞原位进行的核酸分子杂交技术， 敏感度高，特异性强，是当前分子生物学研究的重要手段。 敏感度高，特异性强，是当前分子生物学研究的重要手段。 原位杂交的原理是两条单核苷酸链通过碱基互补原则紧密 结合，形成稳定的杂交体。根据这一原理， 结合，形成稳定的杂交体。根据这一原理，用一条碱基序 列已知、经特定标记的核苷酸链为探针，与组织切片、 列已知、经特定标记的核苷酸链为探针，与组织切片、细 胞制备或染色体标本中的待检DNA或mRNA片段进行杂交， 胞制备或染色体标本中的待检DNA或mRNA片段进行杂交， 然后显示标记物， 然后显示标记物，从而获得待检核酸的分布和含量等信息 (图1-9) 。按照探针分子的性质，可将其分为cDNA探针、 按照探针分子的性质，可将其分为cDNA探针、 cRNA探针和寡核苷酸探针。 cRNA探针和寡核苷酸探针。
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苏木素-伊红染色（HE）
【染色原理】
苏木素染液中带正电荷的蓝色色精与细胞核中带负电荷的脱氧核糖核苷通过
正、负电荷的极性吸附而守成染色；伊红是带负电荷的酸性染料，通过渗透或
弥散作用而使组织着色。
【组织固定与切片】
4%甲醛固定，石蜡切片厚度4~6μm。
【染色用具】
常规染色用具一套：染色皿或染色缸、刻字笔、标签、树脂胶、盖玻片、鸭嘴
镊、绸布或纱布。
滴管数支。
【试剂配制】
1. Harris苏木素液苏木素
2.5g，无水乙醇25ml，钾明矾50g，蒸馏水
500ml，氧化汞1.25g，乙酸20ml。
将苏木素溶于乙醇（加热）后加入预先已加热溶解明矾的蒸馏水中，煮沸
1~2min，改用小火加热，慢慢加入氧化汞（注意防止产生大量气泡和溶液溅
出）再煮沸2min后立即浸于冷水中，冷却后加入乙酸，过滤后使用。
2. 伊红液
（1) 水溶性伊红染液：伊红0.5g，蒸馏水100ml。
（2）醇溶性伊红染液：伊红0.5~1g，80%~95%乙醇100ml。
3. 分化液: 盐酸0.5~1ml，80%乙醇100ml。
4. 返蓝液: 氨水1ml，蒸馏水100ml。
5. 各级乙醇、二甲苯
【注意事项】
1. 石蜡切片在脱蜡前要烘烤使组织与切片粘贴。
烘烤温度50℃，时间30min 或30℃，24h。
2. 脱蜡要彻底，应定期更换脱蜡液。
3. 分化的时间要恰当掌握，可镜下观察着色情况。
4. 透明要充分。
【染色步骤】
1. 石蜡切片二甲苯Ⅰ脱蜡10~15min。
2. 二甲苯Ⅱ脱蜡10~15min。
3. 无水乙醇5min。
4. 95%乙醇5min。
5. 80%乙醇5min。
6. 自来水冲洗1min。
7. 蒸馏水洗1min。
8. 苏木素染液5~15min。
9. 自来水冲洗1~5min。
10. 化液分化数秒至数分钟。
11. 自来水冲洗1~3min。
12. 返蓝液返蓝30s~1min。
13. 自来水冲洗。
14. 伊红液5~10min。
15. 自来水冲洗。
16. 80%乙醇脱水30s。
17. 95%乙醇Ⅰ脱水1min。
18. 95%乙醇Ⅱ脱水1~3min。
19. 无水乙醇Ⅰ脱水2~5min。
20. 无水乙醇Ⅱ脱水2~5min。
21. 二甲苯Ⅰ透明2~5min。
22. 二甲苯Ⅱ透明2~5min。
23. 中性树胶封片。
【结果】
细胞核呈蓝色，软骨组织、钙盐呈深蓝色，黏液呈灰蓝色，细胞质呈粉红色，胶原纤维呈淡粉色，弹力纤维呈亮粉红色，红细胞呈桔红色。