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    K1 - Histotechnic

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    Catherine Elizabet
    The document discusses the process of histology and tissue processing. It describes how cells are studied through methods like tissue fixation, processing, staining and microscopy. The key steps involve fixing tissues, dehydrating them, clearing them, embedding in wax or resin, sectioning and staining to make the cells visible under microscopes. Specialized stains can identify specific cellular components. Histology provides insights into the microscopic structure of tissues.

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    K1 - Histotechnic

    Uploaded by

    Catherine Elizabet
    131 views30 pages
    The document discusses the process of histology and tissue processing. It describes how cells are studied through methods like tissue fixation, processing, staining and microscopy. The key steps involve fixing tissues, dehydrating them, clearing them, embedding in wax or resin, sectioning and staining to make the cells visible under microscopes. Specialized stains can identify specific cellular components. Histology provides insights into the microscopic structure of tissues.

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    HISTOLOGI

    Original Title

    K1- Histotechnic

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    The document discusses the process of histology and tissue processing. It describes how cells are studied through methods like tissue fixation, processing, staining and microscopy. The key steps involve fixing tissues, dehydrating them, clearing them, embedding in wax or resin, sectioning and staining to make the cells visible under microscopes. Specialized stains can identify specific cellular components. Histology provides insights into the microscopic structure of tissues.

    Copyright:

    Attribution Non-Commercial (BY-NC)
    Download as PPTX, PDF, TXT or read online from Scribd
    Download as pptx, pdf, or txt
    131 views30 pages

    K1 - Histotechnic

    Uploaded by

    Catherine Elizabet
    The document discusses the process of histology and tissue processing. It describes how cells are studied through methods like tissue fixation, processing, staining and microscopy. The key steps involve fixing tissues, dehydrating them, clearing them, embedding in wax or resin, sectioning and staining to make the cells visible under microscopes. Specialized stains can identify specific cellular components. Histology provides insights into the microscopic structure of tissues.

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    Attribution Non-Commercial (BY-NC)
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    Download as pptx, pdf, or txt
    of 30

    dr. Alya Amila Fitrie, M.Kes dr.

    Lokot Donna Lubis Department of Histology, Medicine Faculty University of Sumatera Utara Medan 2010

    1 2 3 4

    Explain in general how to study the cell

    Describes methods to study cells with tissue processing and staining, and the benefits of cell and tissue staining

    Describes the various types of microscopes, how microscope works, and the benefits of them

    Explains the terms used in the observation of histological preparations

    Microscopic
    Biochemistry

    Cell culture

    Histology is the study of the microscopic structure of tissue

    Histotechnique

    method of making histological preparations of a particular specimen through a series of processes to become the preparations were ready for analysis.

    Tissue source: Human Animal

    Fiksasi (Fixation);

    Dehidrasi (Dehydration)

    Pembeningan (Clearing)

    Pemotongan Jaringan (Sectioning)

    Pengecoran (Blocking/Casting)

    Pembenaman (Infiltration/Impregn ation/Embedding)

    Pewarnaan (Staining)

    Perekatan (Mounting)

    Pelabelan (Labelling)

    To preserve tissue architecture, tissue must be first be fixed Chemical reagents are used to hold tissue components in their normal places during the subsequent processing They also prevent tissue from rotting and auto-digestion by leaked lysosomes Protective from microbial attack Fixative only preserve protein

    Fixation effect to the processed tissues:


    Inhibit the process of decay and autolisis Tissue preservation Tissue hardening Solidified colloid Optical differentiation Effect of staining

    Things that must be observed in histological tissue fixation is :

    Tissue

    Thickness Tissue part to demonstrate

    Fixation solution

    Volume. Type of fixation solution pH

    Aim

    : Eliminating the water in cells


    Alkohol
    Alkohol 70%, 80%, 90%, masing-masing 1 hari; Alkohol 95% 2 hari ( 2x ganti); Alkohol 100 % 2 hari ( 2x ganti).

    Spiritus atau metanol Sukrosa 20% Aseton

    jarang digunakan tetapi dapat digunakan seperti etanol;

    Sukrosa 20% selama 2 hari;

    3 x 20 menit.

    As

    wax and resin arent miscible with alcohol either, the alcohol is replaced with an organic substances called xylene and the tissue goes transparent

    Substances

    that are used:


    Benzene/ benzol Benzyl benzoat Xylene/xylol

    Chloroform

    Cedar wood oil

    Methyl benzoat

    Embedding tissue provides them with physical supports so they can be sectioned thinly.

    The process involves hardening the tissues by replacing the water with resin or wax.

    Neither resin nor wax miscible with water however, so water is gradually replaced with alcohol in process called dehydration.

    Tools : Histoplate from plastic (casette) and metal plate (mold).

    L shaped piece of metal (Leuckhart) Arrange 2 pieces of the iron pieces of sheet metal to form a

    Spatula for attaching paraffin blocks. Label this tissue identity when blocking.

    cuboid space; Pour a little liquid paraffin at the edges to prevent leaking; Put the tissue; Pour paraffin enough to cover the whole tissue; Avoid air bubble formation.

    For

    optimum viewing, the embedded wax block must be sectioned as finely as possible The ideal thickness of the section would be no wider than the diameter of the cells (57) for light microscopy. Sectioning done by microtome

    The

    thinly cut sections of ribbon are placed onto warm water to flatten them out A glass slide is placed towards the thinly ribbon which picks it up from the water

    Cells are virtually colourless must be stained to be visualised

    Routine stain distinguish the nucleus from the cytoplasm

    Spesific stains : identify spesific cellular, chemical or enzyme components such as carbohydrate.

    Immunohistochemistry (IHC) : identify spesific protein in the cell.

    First process to stain de-waxing

    Most conventional stain are water based and cant mix with wax the wax must be removed from the tissue in order for the stain to penetrate into the tissue
    The process is the reverse of before :
    Section dipped into xylene to remove the wax Dipped into alcohol Dipped into water

    Eosin

    a fluorescent red dye resulting from the action of bromine on fluorescein. It can be used to stain cytoplasm, collagen and muscle fiber for examination under the microscope. Structures that stain readily with eosin are termed eosinophilic.

    Hematoxylin

    a basic dye which binds to nucleic acid (binds to ribosomes and rough endoplasmic reticulum due to DNA/RNA content) it stains them a purple-blue.

    Collagen fiber

    Light microscope Fluorescent microscope Phase contrast microscope Dark field microscope Electron microscope

    To

    be viewed under a microscope cells must be within the microscopes limit or resolution. The rsolving power of an electron microscope (EM) is around 400x of a light microscope (LM).

    The nature and character staining

    Asidofilik, Basofilik

    Shape
    Similarities to other forms the position of something on the polarity of the cells

    Cuboidal, Columnar, spindle, squamous, reticular Star-shaped appearance, Umbrella sign, Ladder like, Signet ring cell Apical, Basal, Centric, Eccentric, Adluminal, Fairy nuclear

    Histology.

    Tissue Processing. Download from www.jameswatts.co.uk Zulham. Bahan kuliah : Dasar-dasar memahami sel dan jaringan. Alberts B et al. 2002. Molecular Biology of the Cell 4th Ed. New York. Garland Science. Young B, Heath JW. Histology A Text and Atlas. 4th Ed. H&E Staining. ABNOVA, http://www.youtube.com/watch?v=2D0rj0m6 dVs

    Thank you

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