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所在平台: Udemy |
课程主页: https://www.udemy.com/course/histology-step-by-step/
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此课程“医疗学生的组织学基础”旨在为医学学生提供组织学(也称为微观解剖学)的全面介绍。组织学是研究生物组织微观解剖学的学科,与不使用显微镜的宏观解剖学互补。虽然微观解剖学可以细分为器官学(研究器官)、组织学(研究组织)和细胞学(研究细胞),但现代用法将所有这些主题都归入组织学领域。在医学领域,组织病理学是组织学的一个分支,涉及疾病组织的微观识别和研究。 课程深入探讨了动物(包括人类)的四种基本组织类型: * **上皮组织(Epithelium):** 包括单层上皮(如鳞状、立方、柱状和假复层柱状)和复层上皮(如复层鳞状、复层立方、复层柱状和移行上皮)。还涵盖了多细胞腺体。 * **肌肉组织(Muscle tissue):** 细分为平滑肌、骨骼肌和心肌。 * **结缔组织(Connective tissue):** 包括一般结缔组织(如疏松和致密结缔组织)和特殊结缔组织(如软骨、骨、造血组织和血液、淋巴)。 * **神经组织(Nervous tissue):** 涵盖中枢神经系统和周围神经系统,以及特殊感受器。 课程强调了用于观察这些组织类型的各种组织学染色技术。它解释了染色剂(如苏木精和伊红)如何突出组织的关键特征和成分,以及为何需要多种染色剂来全面了解样本。此外,课程还提到了特殊的染色剂,它们可以特异性地突出特定蛋白质,但可能无法显示其他结构。 **组织制备**是关键步骤,确保了后续染色的准确性。此过程包括: * **固定(Fixation):** 使用化学物质(如中性缓冲福尔马林)保存组织结构并防止降解。 * **脱水(Dehydration):** 使用乙醇去除水分,并进一步硬化组织,为显微镜检查做准备,随后使用二甲苯去除乙醇。 * **包埋(Embedding):** 将样本放入石蜡或树脂中,以利于提取细胞结构,但需注意石蜡可能会影响免疫染色。 * **切片(Sectioning):** 使用显微切片机将组织切成薄片(通常为 4-5 微米),以便染色和在显微镜载玻片上进行检查。 * **抗原恢复(Antigen Retrieval):** 在固定和包埋过程中,某些抗原可能会被掩盖。此步骤通过加热或蛋白酶处理来恢复这些抗原,提高免疫染色效果,尽管存在使固定剂和抗原变性的风险。 课程还提到了**活体染色**,这是一种在组织保留在体内时使用的特殊染色技术,对于识别特定组织类型和异常组织,从而提高活组织检查的准确性非常重要。 总而言之,该课程为医学学生提供了组织学的基本原理,包括不同组织类型的结构、相关的染色技术以及高质量组织样本制备的必要步骤。
Histology, also known as microscopic anatomy or microanatomy, is the branch of biology that studies the microscopic anatomy of biological tissues Histology is the microscopic counterpart to gross anatomy, which looks at larger structures visible without a microscope.[5][6] Although one may divide microscopic anatomy into organology, the study of organs, histology, the study of tissues, and cytology, the study of cells, modern usage places all of these topics under the field of histology. In medicine, histopathology is the branch of histology that includes the microscopic identification and study of diseased tissue.There are four basic types of animal tissues: muscle tissue, nervous tissue, connective tissue, and epithelial tissues All animal tissues are considered to be subtypes of these four principal tissue types (for example, blood is classified as connective tissue, since the blood cells are suspended in an extracellular matrix, the plasma)EpitheliumSimple epitheliumSimple squamous epitheliumSimple cuboidal epitheliumSimple columnar epitheliumPseudostratified columnar epitheliumStratified epitheliumStratified squamous epitheliumStratified cuboidal epitheliumStratified columnar epitheliumTransitional epitheliumMulticellular glandsMuscle tissueSmooth muscleSkeletal muscleCardiac muscleConnective tissueGeneral connective tissueLoose connective tissueDense connective tissueSpecial connective tissueCartilageBoneHemopoieticBloodLymphNervous tissueCentral nervous systemPeripheral nervous systemSpecial receptorsFour basic types of human tissue can be stained and viewed using various histological techniques. Epithelium, connective tissue, muscle tissue, and nervous tissue have commonalities but look very distinct structurally after staining. Each stain exists to highlight an important feature or component within a tissue type. For example, one of the most common stains, Hematoxylin, is a basic dye that stains proteins a blue color, while Eosin stains proteins a pink color. These two stains are commonly used together to define intracellular organelles and proteins. Because of the variety of the proteins that exist, some stains were created to highlight a particular protein, which this review will discuss in the following sections. The benefit of using a special stain is that it can highlight the specific protein very well. However, because of its specificity, the other structures will not be seen. For this reason, multiple slides will often be created from a given specimen so that multiple stains can be performed to gather the full range of needed information.Almost all tissue stains are performed on tissue that has been removed from the body. However, in rare instances, very specialize stains called vital stains can work on tissue remaining in the body. These stains are used for the identification of specific types of tissue and identification of abnormal tissue, so a subsequent biopsy can be more accurate in obtaining abnormal tissueTissue PreparationBefore specific staining can occur, tissue samples must undergo preparation through the following stages: Fixation, processing, embedding, sectioning, and sometimes antigen retrieval. In modern histology laboratories, most of these steps are automated..Fixation: Fixation uses chemicals to preserve the structure of the tissue in its natural form and protects it from degradation by irreversibly cross-linking proteins. Although several specialized fixatives are available, Neutral Buffered Formalin is a common choice for this step. The fixation step is vital to the rest of the histologic staining procedure because by retaining the chemical composition of the tissue, the sample is hardened and makes the sectioning phase easier. Paraffin-formalin is another effective fixative. Its benefit is that it is the fixative of choice for immunostaining; however, it requires preparation at the time of the fixation. Bouin is a fixative used for examining embryo and brain tissue because of its superior preservation of delicate nuclei and glycogen. Its downside is that it does not preserve kidney tissues well and also distorts mitochondrial structure.[1]Dehydration: The addition of ethanol accomplishes the dehydration of a sample. It removed water from the sample and further hardens the tissue for eventual light microscopy. After ethanol is applied, and following the completion of tissue dehydration, xylene is used to remove the ethanol.[1]Embedding: Embedding is the process of putting the sample into a paraffin wax or a plastic resin to enhance the process of extracting cellular structures. This step is to be performed with caution if the goal is to perform immunostaining because the paraffin wax will inhibit the penetration of antibodies, and lead to a false result.[1]Sectioning: Sectioning involves mounting the specimen on a microtome and cutting it into sections. The preferred thickness is 4-5 micrometers so that it can be stained and put on a microscope slide for examination.[1]Antigen Retrieval: This step is to retrieve antigens that could have been covered in the fixation and embedding stages. If the cross-linking of proteins conceals the antigen sites, there may not be as robust of an immunohistochemical response. Antigen retrieval is achieved through heating and proteolytic methods to break down the cross-links and reveal the epitopes and antigens that were previously covered.[1] Although this step carries the risk of denaturing both the fixative and the antigens themselves, a successful antigen retrieval method can lead to a much more effective immunostaining intensity.