Dr Vivien Rolfe De Montfort University This is an Open Educational Resource (OER) that is globally available on the web Creative Commons BY SA.

Slides:



Advertisements
Similar presentations
1 Cells & Organelles Quick Quiz 3 Press the button when you’re sure… remember to take your time as you only get one chance!
Advertisements

On reaching this century and with the ease of information flow, the world seemed to be living in a global village full of a complicated information. We.
381 PHT Lab #2.
S T A I N I N G DWI WINARNI Departemen Biologi Fakultas Sains dan Teknologi Universitas Airlangga Surabaya.
Dr Vivien Rolfe De Montfort University This is an Open Educational Resource (OER) that is globally available on the web Creative Commons BY SA.
Classification of Bacteria. There are thousands of species of bacteria on earth, many of which have not yet been identified. When attempting to classify.
Copyright © 2006 Pearson Education, Inc., publishing as Benjamin Cummings PowerPoint ® Lecture Slide Presentation prepared by Christine L. Case M I C.
BACTERIAL STAINING.
Experimental pathology refers to the observation of the effects of manipulations on animal models or cell cultures regarding researches on human diseases.
© red ©
Dr Vivien Rolfe De Montfort University This is an Open Educational Resource (OER) that is globally available on the web Creative Commons BY SA.
Dr. Samah Kotb Nasr Eldeen. Biological stains are prepared from dyes which have been manufactured to rigid specifications; to ensure that they are suitable.
CYTOLOGY BY Dr. TAREK ATIA Histology and Cell Biology.
Staining HAEMATOXYLIN & EOSIN
Histological Stains. Tissue Processing To some extent, light-level (as opposed to e.m.- level) histology is a lost art, especially if one is interested.
PATHOPHYSIOLOGY PATHOPHYSIOLOGY w DEFINED Involves the study of function that results from disease processes.
Special stain 2.
The Size of It All Types of Microscopes. The Size of It All Remember that 1 inch = 2.54 cm and that 1 meter contains micrometers (µm) or
Staining of Bacteria Staining of Bacteria Brno University of Technology, Faculty of Chemistry Hana ŠURANSKÁ.
Third lecture for microbiology manual
+ Research Techniques I (Biology 513) Staining. + Why stain tissue? Most tissues are colorless and unless one uses diffraction interference contrast microscopy,
1 Bio211 Laboratory 2 Epithelial and Connective Tissues.
Dr. Samah Kotb 2015 Histology Techniques CLS 322.
Bacterial Staining General Discussion. Stains All dyes are salts –Ionize Cationic Anionic Techniques –Single dyes –Multiple dyes Use.
S TAINING M ICROORGANISMS An overview of staining.
Cell Biology Cell Biology S. Rahgozar,PhD University of Isfahan Faculty of Science 3. Vital and non-vital staining 1.
CHAPTER 3 Observing Organisms Through a Microscope Units of Measurements Microscopy: The Instruments Preparation of Specimens.
Methods in histology Microscopy Maňáková Content of practices Organization of the practices How microscopic slides are made Microscope Staining.
SEPARATION AND DETECTION OF PROTEINS Part I Jana Vobořilová, Anna Kotrbová-Kozak, Vlasta Fürstová, Tereza Kopská.
Tissue Practice Test Dr. B. 1. Identify the tissue.
Connective tissue (1) 1.Loose connective tissue (No.4, No.1 )
STAINING.
Vessels Digital Laboratory
PHT 226 Lab # 3 Gram’s stain (mixture) Acid fast stain Spore stain.
Lab 4 & 5 Staining Technique
PREPARATION OF A SMEAR GRAM STAIN ACID FAST STAIN
Week : 4.ERT 144 staining (rev, quiz) isolation & identification.
INTRODUCTION TO HISTOLOGY
Electron Microscopy 7th lecture.
Special Staining (I).
Observing Microorganisms through a Microscope Dr. Bhavesh Patel Principal V.P. and R.P.T.P. Science College Vallabh Vidyanagar –
Gram Staining. Aseptic Technique Sterilize Everything – Use bunsen burner or Bacti-Cinerator.
Inorganic Analysis Fish!. Pick a question!
Introduction to Muscle Tissue. Skeletal Muscle - red fibres - white fibres - intermediate fibres Cardiac Muscle - cardiomyocytes - Purkinje fibres Smooth.
Stains and Staining Reactions. Stains and objectives of staining Stains are useful for the following reasons It makes the microscopic semi-transparent.
 The sections, as they are prepared, are colourless and different components cannot be appreciated.  Staining them by different coloured dyes, having.
Introduction to pathology & causes of diseases
Histological techniques: Haematoxylin and Eosin staining
Elastica van Gieson-picro-fuchsin staining
Biology 131 – Anatomy and Physiology Instructor: Ruth Williams
Lab 4 & 5 Staining Technique
Principles of Laboratory Diagnosis of Infectious Diseases
SPECIAL STAINS FOR NUCLEIC ACID.
HISTOLOGY.
Examples of different types of connective tissues. A
THE UTILITY OF AZAN TRICHROME STAINING IN AMELOBLASTOMA
Staining Methods.
The Study of Microbial Structure: Microscopy and Specimen Preparation
Why we should be Stain Bacteria
Bright-Field Microscopy
Gram-Positive Bacteria
Tissue Structure and Function
Introduction and Epithelium
Inorganic Analysis Fish!.
Colours.
Lab 4 & 5 Staining Technique
Tissue Structure and Function
140MIC: Microbiology Lecture-6 Microscopes.
Isaac Perea-Gil et al. BTS 2016;1:
Indicators.
Presentation transcript:

Dr Vivien Rolfe De Montfort University This is an Open Educational Resource (OER) that is globally available on the web Creative Commons BY SA

 Why are special stains still important and relevant today?  What are some of the chemical principles behind these stains?  Some common examples that can be prepared in student laboratory teaching.

 H&E was first introduced in the 1870’s and the term special stain came to refer to any technique other than H&E used in the clinical environment.  Whilst the H&E stain is the most common staining method in hospital and research laboratories, it isn’t without its limitations. H&E cannot visualize micro-organisms. H&E is not good for distinguishing connective tissue and nerve tissue. H&E cannot distinguish molecular basis of disease and immunohistochemistry might be preferred.

 Tolonium Chloride  Useful blue cationic dye  Cheap and simple application

 Immunohistochemical methods have advanced but are costly and reagents deteriorate quickly.  Special stains include silver methods (such as Gordon and Sweet’s), gold, or Luxol fast blues to stain myelin.  Other special stains identify nerve cells. The techniques are important for looking and neurodegeneration.

 One of several silver methods for staining reticulin.  Tissue treated with potassium permanganate to enable the silver to bind.  Uses an ammoniacal silver solution.  What is reticulin?  Why is it important?

Liver tissue with no counterstain Reticulin = black

Liver counterstained with what? Reticulin = black Cytoplasm = pink

 Trichrome stain – producing 3 colours.  Anionic dye and a cationic counterstain.  Nuclear stain applied first such as Weigert’s haematoxlin.  Collagen stains red with acid fuchsine.  Cytoplasm including muscle stains yellow.  Washing in acidified water differentiates tissue producing two colours.

Bladder Collagen = red All other tissue including transitional epithelium = yellow

Collagen = red Smooth muscle = yellow Epithelium = yellow Application? Might be used to localise tumours in the bladder to either the smooth muscle or connective tissue layers.

 Trichrome stain.  Martius yellow and phosphotungstic acid.  Brilliant crystal scarlet.  Methyl blue.  What do the dyes stain?

Epithelium = red Collagen = blue Cytoplasm = red No visible yellow

Collagen = blue Erythrocytes and early fibrin = yellow Cytoplasm = pink

Erythrocytes clearly yellow Collagen = blue Cytoplasm = red

Early fibrin deposits = diffuse yellow staining Collagen = blue Glandular tissue = red

 Trichrome stain.  Iron-haematoxylin plus two anionic dyes.  MSB is a variation of this.  Iron-haematoxylin.  Scarlet-acid fuchsine.  Light green (more of a turquoise stain).

Nuclei = black Cytoplasm including muscle and epithelium = red Erythrocytes = red Collagen = bluey green or turquoise

MSB Collagen = bluey green Cytoplasm of epithelium and skeletal muscle = red

 Histological and Histochemical Methods. 4 th Edition. By JA Kiernan Scion publishing. Available:  Histopathology: Fundamentals of Biomedical Science. By G Orchard and B Nation Oxford University Press. Available:  Laboratory skills open educational resources. De Montfort University. Available: