TISSUE PREPARATION.

Slides:



Advertisements
Similar presentations
Gross Techniques In Surgical Pathology. Introduction The routine work associated with a surgical pathology specimen includes gross & microscopic examinations.
Advertisements

PATHOPHYSIOLOGY PATHOPHYSIOLOGY w DEFINED Involves the study of function that results from disease processes.
Tissues of the Body Module (TOB) Introductory Lecture LIGHT MICROSCOPY.
Types of microscopes & Microtechniques.
Artifacts in Histological Preparations. Microwave heating It’s a kind of low power energy carried by micro-waves Internal direct heating is our advantage.
Immunohistochemistry
+ Research Techniques I (Biology 513) Tissue processing.
Dr. Samah Kotb Nasr Eldeen.  Several types of staining processes are used to color tissues for microscopical examination.  Staining methods depend.
PREPARATION OF HISTOLOGICAL SPECIMENS
Tissue Processing.
Sectioning or Microtomy
CYTOLOGY BY Dr. TAREK ATIA Histology and Cell Biology.
PATHOPHYSIOLOGY PATHOPHYSIOLOGY w DEFINED Involves the study of function that results from disease processes.
Introduction to Histology Tissue processing and Microscope
Histology and Embryology 组织学与胚胎学 Department of Histology and embryology Three Gorges University, Yichang, China.
+ Research Techniques I (Biology 513) Staining. + Why stain tissue? Most tissues are colorless and unless one uses diffraction interference contrast microscopy,
1- Histology and Histo-technique
Histology Histology is the study of the tissues of the body and how these tissues are arranged to constitute organs Literally, histology means tissue or.
Dr. Samah Kotb 2015 Histology Techniques CLS 322.
Decalcification. Decalcification are the most type here, but other tissues may contain calcified areas as well. Bone specimens are the most type here,
Tissue Processing Dr : Hala El-sayed Mahmoud
PREPARATION OF HISTOLOGICAL SPECIMENS
A laboratory guide for histology 刘尚明 武玉玲. Introduction  As other medical courses, the study of histology consists of two parts: lectures and laboratory.
Histology and Embryology
The Process Of Molecular Cytology: Embedding and Sectioning Natasha Williams Dr. Katia Manova Zuckerman Research Building/ MSKCC.
组织胚胎学课件 七年制英文医学班专用 中国医科大学 基础医学院 组胚 — 英文教学组. Chapter 1 Introduction.
Histology Techniques CLS 322
Procedures Fixation Tissues must be immersed in fixative immediately after removal from the body . 10% Neutral Buffered Formalin is the routine fixative.
+ Research Techniques I (Biology 513) Fixation. + Introduction Why do we fix tissue What makes an ideal fixative? Penetrate rapidly and prevent postmortem.
Light Microscope. Light Microscope Light Microscope The light microscope depends on light passing through an object in order for it to be seen. The result.
Starter: Microscopes Which image is from the light microsope? How do you know?
Histology & Its Methods of Study 2015/16 1Lufukuja G.
Histology and Embryology Zhong jie Li (李仲杰) School of medicine, Zhejiang University
SELECTION OF THE TISSUE BLOCK
Lab 2: Tissue Processing
Course Descriptive Histology Introduction.
Introduction.
PRACTICAL -1 PREPARATION OF THE FIXATIVE. HISTOLOGICAL FIXATIVES.
PRACTICAL -2 TISSUE PROCESSING 1 Dr.Nessrin Alabdallat.
Preparation of tissues for study
Electron and Light Microscopes Revision Scientific Detection.
Preparation of Plant tissues for histological study
Introduction to Muscle Tissue. Skeletal Muscle - red fibres - white fibres - intermediate fibres Cardiac Muscle - cardiomyocytes - Purkinje fibres Smooth.
Light Microscope Terms and Practices.
Introduction to Histology Light and Electron Microscopes
Pathophysiology.
“HANDS ON TRAINING ON BIOLOGICAL TECHNIQUES”
Histological techniques: Haematoxylin and Eosin staining
Histology introduction
Histological Techniques
Pathophysiology.
بسم الله الرحمن الرحيم Department of Pathology College of Medicine
PATHOPHYSIOLOGY.
Biotechnique (BIOL 410) Histology.
SPECIAL STAINS FOR NUCLEIC ACID.
بسم الله الرحمن الرحيم Department of Pathology College of Medicine
College of Education Biology Dept.
INTRODUCTION TO HISTOLOGY
Staining Methods.
Topic 1: Introduction to Histology
Published in Thyroid. January 2014, 24(1): DOI: /thy
Microscopic study include: 1. Study of oral soft tissues Microscopic examination is the method used to study the histological structure of the oral.
General Principles of Tissue Preparation and Staining
Keep & Study in ISN for Quiz next class
Pathophysiology.
Methods of Sample Preparation
Histology and Embryology
Immunohistochemistry
Interpretation of Histological sections
Tissue processing Histology:
Presentation transcript:

TISSUE PREPARATION

Tissue fixation: Tissue is treated with fixatives. 4% Formaldehyde or physical fixative as heat or microwave. These fixatives causes crosslinking of macromolecules so that biological Activity is arrested. Tissue embedding Fixed tissues are cut in small slices.for this we have to embed them in Parrafin wax for light microscopy Resin for electron microscopy.

As tissues are hydrophilic and wax is hydrophobic what to do now Dehydrate tissues with ascending grades of alcohol 70 to 100% Dehydrating agent was relpaced by immersing in Xylene. These two processes are called dehydration and clearing . Now tissue ready for embedding. Tissues placed in metallic mould filled with paraffin wax at 58-60 degree. Heat evaporates xylene and tissues get paraffinized. Tissue block placed in microtome for cutting. An instrument for slicing of tissues usaually 4-5µm.

STAINING OF SLIDES: Usually H & E stains were used. They are water based stain and tissue is embedded in wax. Now what to do Put XYLENE and then rehydrate it. Descending grades of alcohol and then with distilled water Put hematoxyllin and decolorize it with acid alcohol quick dips and checked Microscopically. Acidic structures stain this basic dye. Rinse with tap water .Put eosin now Basic structures stain this acidic dye.

PRESERVE THE SLIDE: Dehydrate again with ascending grades of alcohol And cleared with xylene. Mount with coverslip.

Results: Nuclei Blue/Black Cytoplasm Varying shades of pink Red blood cells Orange/red

Artifacts in histology

  What one may see upon microscopic examination of sections of specimens of human and animal tissues is not always related to the normal histology or pathology of the tissue in question. Defects or abnormalities in tissue sections are referred to as artifacts. Structures or features in tissue that interfere with normal histological examination.

Artifacts may occur at different stages in the routine collection of tissues, fixation, processing, cutting and staining of tissues. Not always present in normal tissue and can come from outside sources. Interfere with histology by changing the tissues appearance and hiding structures.

Pre-histology Features and structures introduced prior to the collection of the tissues e.g. ink from tattoos.

Post-histology Artifacts can result from tissue processing. Processing commonly leads to changes like shrinkage, color changes in different tissues types and alterations of the structures in the tissue. Because these are caused in a laboratory the majority of post histology artifacts can be avoided or removed after being discovered.

Formalin pigment

Tissue Fold

Tissue Tear

Retraction artifact

Unfixed tissues

Rough or Rapid Cutting

Failure To Stain Because Of Residual Wax On The Section

Floater/Carry over