SlideShare a Scribd company logo
Hematoxylin & Eosin Stain
Part-1 of Microscopic tissue fixation techniques used in pharmacological studies
Priyansha Singh
B. Pharm, M.S. (Pharm.)- Pharmacology & Toxicology
The sections of tissues as they are prepared are colourless & different components
cannot be identified. Staining makes them differently coloured as various dyes have
affinities towards specific components of tissues, making the identification &
morphological studies possible. Hematoxylin & Eosin- most frequently used stain
in histology
INTRODUCTION
• Hematoxylin and eosin staining technique functions to recognize different types of tissues and their
morphological changes, especially in cancer diagnosis.
• It is mainly used as a gold standard stain in medical diagnosis
• Hematoxylin has a deep blue-purple color and stains nucleic acids.
• Eosin is pink and stains proteins non-specifically.
• In a typical tissue, nuclei are stained blue, whereas the cytoplasm and extracellular matrix have varying
degrees of pink staining.
• Hematoxylin and eosin are both dyes, and naturally, dyes have a high affinity for tissues, depending on the
Acidity and/or alkalinity of the dyes.
OBJECTIVE
To demonstrate the morphologies of tissues and cells
PRINCIPLE
• Haematoxylin is a dye which when combined with a mordant like potash alum starts to behave like a basic/ cationic
dye. The staining of nucleus is blue due to binding of dye metal complex with the nuclear material.
• Eosin dye is acidic dye hence it as a negative charge (eosinophilic). Therefore it stains the basic protein structures of
a cell & extracellular matrix (acidophils or eosinophils), giving them a red or pink color, for example, the cytoplasm
is positively charged, and therefore it will take up the eosin dye, and appear pink.
Main types of staining seen on H&E stain
Unstained tissue section
PROCEDURE
Materials Required
• Xylene I & II- for deparaffinization & clearance
• 70%, 80%, 100% alcohol- for rehydration & dehydration after counter staining
• Haematoxylin- for nuclear staining
• 1% acid alcohol- for differentiation
• Li2CO3- for bluing
• Eosin- counter staining
• DPX- mounting
Procedure with principle behind it
• Deparaffinization- to remove wax embedded in the tissue.
• Descending grades of alcohol- After thorough de-waxing, the slide is passed through several changes of
alcohol to remove the xylene, then thoroughly rinsed in water. The section is now hydrated so that aqueous
reagents will readily penetrate the cells and tissue elements.
• Haematoxylin- for nuclear staining
• Differentiation- to remove non-specific background staining and to improve contrast.
• Bluing- changes the color of hematoxylin from red to blue by chemically altering the dye with a slightly
basic solution. The bluing solution's alkaline pH stabilizes the stain and makes it insoluble in water and
alcohol. The change in pH is what causes the color change.
• Counterstaining- A counterstain is a stain with color contrasting to the principal stain, making the stained
structure easily visible using a microscope.
• Ascending grades of alcohol- for dehydration. To remove water from the tissue so that paraffin wax can
completely penetrate it. Since wax is not water soluble, but is soluble in xylene, the water in the tissue must
be replaced with xylene.
• Clearance by Xylene I & II- to remove dehydrating agents and prepare tissues for impregnation with the
embedding agent. Clearing makes the tissue transparent, or clear, making the slides easier to read.
• DPX mounting- to increase the refractive index and improve image quality in microscope
RESULTS
• Nuclei are stained blue, cytoplasm & ECM- varied degrees of pink stain
• Well fixed cells/ tissues show considerable intracellular details
• Nuclei stains with eosin
• If polyribosomes are present then cytoplasm will have a distinct blue cast
Masson’s Trichrome Staining
Part-2 of Microscopic tissue fixation techniques used in pharmacological studies
Priyansha Singh
B. Pharm, M.S. (Pharm.)- Pharmacology & Toxicology
Definition
• Masson’s trichrome is a differential staining technique for selective
staining of collagen fibres, fibrin, muscles & RBCs.
• Trichrome stain- Wiegert's haematoxylin (nucleus)
- Biebrich scarlet (muscle, cytoplasm & nucleus)
- Aniline/ methylene blue/ light green (collagen)
Objectives of Masson’s Trichrome staining
• To stain the collage fibers.
• To stain collagen.
• To stain keratin.
• To stain fibrin.
• To stain muscle fibers.
• To differentiate between collagen & smooth muscle in tumors, increase in collagen in diseases like
cirrhosis. Routine stain in kidney fibrosis & liver cirrhosis biopsies.
Principal behind the staining
• As the name implies, 3 dyes are employed selectively
staining muscle, collagen fibers, fibrin & erythrocytes.
• The general rule in trichrome staining is that the less
porous tissues are coloured by the smallest size dye
molecule.
• Wherever a dye of large molecular size is able to
penetrate but the dye of smaller size passes off the
larger pore size.
• The tissue is first stained with the acid dye (Biebrich
Scarlet red- Acid Fuchsin) binding with acidophilic
tissue components.
• Further treatment with the phospho acids, the less
permeable components retain red stain and the red
colour comes out of the collagen pores
• Collagen gets bind with aniline blue.
Fix the sample using Bouin’s
solution- mordant (microwave
for 1 min) stand for 15 mins
Add Wiegert's
Haematoxylin to stain
nucleus (10 mins)
Add Biebrich scarlet red
stain (5 mins) to stain
cytoplasm, keratin, muscle
& RBCs
Add Phosphotungstic/
phosphomolybdic acid for
decolorization (10 mins).
Add aniline blue/ methylene
blue/ light green + 1% acetic acid
to stain collagen
Wash (5 mins)
Wash & rinse
Rinse
Procedure of staining
Results
Nuclei black
Cytoplasm, Muscle, Erythrocytes- red
Collagen- blue
ADVANTAGES
1. To measure & differentiate collagen deposition as compared to
HE staining
2. Produces clear images of collagen compared to other stains
3. More accurate than other staining techniques
DISADVANTAGES
1. Preparation of solutions to be used within 24 hrs
2. Corrosive stains
Sirius red staining
Part-3 of Microscopic tissue fixation techniques used in pharmacological studies
Priyansha Singh
B. Pharm, M.S. (Pharm.)- Pharmacology & Toxicology
Histopathological staining techniques used in liver diseases
Histopathological staining techniques used in liver diseases
Histopathological staining techniques used in liver diseases
Histopathological staining techniques used in liver diseases

More Related Content

Similar to Histopathological staining techniques used in liver diseases

Staining of histological slides.pptx
Staining of histological slides.pptxStaining of histological slides.pptx
Staining of histological slides.pptx
NoorKhan188957
 
Vital staining ppt notes
Vital staining ppt notesVital staining ppt notes
Vital staining ppt notes
Anil Yadav
 
Histopathology Embedding to Staining the Slides
Histopathology Embedding to Staining the SlidesHistopathology Embedding to Staining the Slides
Histopathology Embedding to Staining the Slides
Rukhshanda Ramzaan
 
Histological techniques
Histological techniquesHistological techniques
Histological techniques
RUPALIMUNDE
 
Special stains
Special stainsSpecial stains
Special stains
imrana tanvir
 
Hematoxylin and eosin staining Dr.Abhinav Golla MD Pathology
Hematoxylin and eosin staining Dr.Abhinav Golla MD PathologyHematoxylin and eosin staining Dr.Abhinav Golla MD Pathology
Hematoxylin and eosin staining Dr.Abhinav Golla MD Pathology
Aadhya Medicure Pathlabs Vijayanagar colony.hyderabad
 
Histology
Histology Histology
Histology
sania aslam
 
Massons Trichrome Stain
Massons Trichrome StainMassons Trichrome Stain
Massons Trichrome Stain
Zahoor Ahmed
 
Lecture 1.introduction to histopathology ii
Lecture 1.introduction to histopathology iiLecture 1.introduction to histopathology ii
Lecture 1.introduction to histopathology ii
Catholic university of Rwanda
 
H and E staingin.ppt
H and E staingin.pptH and E staingin.ppt
H and E staingin.ppt
Pravin Amabade
 
staining technique.pptx
staining technique.pptxstaining technique.pptx
staining technique.pptx
HimadriSekharMukherj
 
Histological Tools
Histological Tools Histological Tools
Histological Tools
Tejinder Pal Singh
 
STAINING._110459.pptx
STAINING._110459.pptxSTAINING._110459.pptx
STAINING._110459.pptx
ssuserf31797
 
Histopathology.pptx
Histopathology.pptxHistopathology.pptx
Histopathology.pptx
WondwossenAfrash
 
Staining seminar 2
Staining seminar 2Staining seminar 2
Staining seminar 2
dentist at the making
 
ANATOMY ELECTIVE HAEMAATOXYLIN AND EOSIN
ANATOMY ELECTIVE HAEMAATOXYLIN AND EOSINANATOMY ELECTIVE HAEMAATOXYLIN AND EOSIN
ANATOMY ELECTIVE HAEMAATOXYLIN AND EOSIN
seervidivyanshu18869
 
Notes for STAINING AND ANALYSIS of HISTOLOGICAL PREPARATIONS
Notes for STAINING AND ANALYSIS of HISTOLOGICAL PREPARATIONSNotes for STAINING AND ANALYSIS of HISTOLOGICAL PREPARATIONS
Notes for STAINING AND ANALYSIS of HISTOLOGICAL PREPARATIONS
improvemed
 
Connective tissue
Connective tissueConnective tissue
Connective tissue
Shabab Ali
 
Introduction to histology
Introduction to histologyIntroduction to histology
Introduction to histology
Abdul Ansari
 
Histopatholgy staining by suchit kumar
Histopatholgy staining by suchit kumarHistopatholgy staining by suchit kumar
Histopatholgy staining by suchit kumar
suchitkumar24
 

Similar to Histopathological staining techniques used in liver diseases (20)

Staining of histological slides.pptx
Staining of histological slides.pptxStaining of histological slides.pptx
Staining of histological slides.pptx
 
Vital staining ppt notes
Vital staining ppt notesVital staining ppt notes
Vital staining ppt notes
 
Histopathology Embedding to Staining the Slides
Histopathology Embedding to Staining the SlidesHistopathology Embedding to Staining the Slides
Histopathology Embedding to Staining the Slides
 
Histological techniques
Histological techniquesHistological techniques
Histological techniques
 
Special stains
Special stainsSpecial stains
Special stains
 
Hematoxylin and eosin staining Dr.Abhinav Golla MD Pathology
Hematoxylin and eosin staining Dr.Abhinav Golla MD PathologyHematoxylin and eosin staining Dr.Abhinav Golla MD Pathology
Hematoxylin and eosin staining Dr.Abhinav Golla MD Pathology
 
Histology
Histology Histology
Histology
 
Massons Trichrome Stain
Massons Trichrome StainMassons Trichrome Stain
Massons Trichrome Stain
 
Lecture 1.introduction to histopathology ii
Lecture 1.introduction to histopathology iiLecture 1.introduction to histopathology ii
Lecture 1.introduction to histopathology ii
 
H and E staingin.ppt
H and E staingin.pptH and E staingin.ppt
H and E staingin.ppt
 
staining technique.pptx
staining technique.pptxstaining technique.pptx
staining technique.pptx
 
Histological Tools
Histological Tools Histological Tools
Histological Tools
 
STAINING._110459.pptx
STAINING._110459.pptxSTAINING._110459.pptx
STAINING._110459.pptx
 
Histopathology.pptx
Histopathology.pptxHistopathology.pptx
Histopathology.pptx
 
Staining seminar 2
Staining seminar 2Staining seminar 2
Staining seminar 2
 
ANATOMY ELECTIVE HAEMAATOXYLIN AND EOSIN
ANATOMY ELECTIVE HAEMAATOXYLIN AND EOSINANATOMY ELECTIVE HAEMAATOXYLIN AND EOSIN
ANATOMY ELECTIVE HAEMAATOXYLIN AND EOSIN
 
Notes for STAINING AND ANALYSIS of HISTOLOGICAL PREPARATIONS
Notes for STAINING AND ANALYSIS of HISTOLOGICAL PREPARATIONSNotes for STAINING AND ANALYSIS of HISTOLOGICAL PREPARATIONS
Notes for STAINING AND ANALYSIS of HISTOLOGICAL PREPARATIONS
 
Connective tissue
Connective tissueConnective tissue
Connective tissue
 
Introduction to histology
Introduction to histologyIntroduction to histology
Introduction to histology
 
Histopatholgy staining by suchit kumar
Histopatholgy staining by suchit kumarHistopatholgy staining by suchit kumar
Histopatholgy staining by suchit kumar
 

More from PHARMA IQ EDUCATION

Immunopharmacology & immunology Unit-1 (PC-660)
Immunopharmacology & immunology Unit-1  (PC-660)Immunopharmacology & immunology Unit-1  (PC-660)
Immunopharmacology & immunology Unit-1 (PC-660)
PHARMA IQ EDUCATION
 
Pain Models-Screening Assay in vitro & in vivo
Pain Models-Screening Assay in vitro & in vivoPain Models-Screening Assay in vitro & in vivo
Pain Models-Screening Assay in vitro & in vivo
PHARMA IQ EDUCATION
 
Caco-2 cell permeability assay for drug absorption
Caco-2 cell permeability assay for drug absorptionCaco-2 cell permeability assay for drug absorption
Caco-2 cell permeability assay for drug absorption
PHARMA IQ EDUCATION
 
Acute Poisoning and it's management.pptx
Acute Poisoning and it's management.pptxAcute Poisoning and it's management.pptx
Acute Poisoning and it's management.pptx
PHARMA IQ EDUCATION
 
BIOTRANSFORMATION UNIT -1 DRUG METABOLISM (1).pptx
BIOTRANSFORMATION UNIT -1 DRUG METABOLISM (1).pptxBIOTRANSFORMATION UNIT -1 DRUG METABOLISM (1).pptx
BIOTRANSFORMATION UNIT -1 DRUG METABOLISM (1).pptx
PHARMA IQ EDUCATION
 
Significance of DMPK in drug discovery.pptx
Significance of DMPK in drug discovery.pptxSignificance of DMPK in drug discovery.pptx
Significance of DMPK in drug discovery.pptx
PHARMA IQ EDUCATION
 
CONTAMINANTS IN CELL CULTURE & PRECAUTIONS.pptx
CONTAMINANTS IN CELL CULTURE & PRECAUTIONS.pptxCONTAMINANTS IN CELL CULTURE & PRECAUTIONS.pptx
CONTAMINANTS IN CELL CULTURE & PRECAUTIONS.pptx
PHARMA IQ EDUCATION
 
UNIT- 10 ANIMAL CELL CULTURE TECHNIQUES (3).pptx
UNIT- 10 ANIMAL CELL CULTURE TECHNIQUES (3).pptxUNIT- 10 ANIMAL CELL CULTURE TECHNIQUES (3).pptx
UNIT- 10 ANIMAL CELL CULTURE TECHNIQUES (3).pptx
PHARMA IQ EDUCATION
 
SUMMARY OF MEDIA USED IN CELL CULTURE.docx
SUMMARY OF MEDIA USED IN CELL CULTURE.docxSUMMARY OF MEDIA USED IN CELL CULTURE.docx
SUMMARY OF MEDIA USED IN CELL CULTURE.docx
PHARMA IQ EDUCATION
 
SUMMARY OF MEDIA USED IN CELL CULTURE.docx
SUMMARY OF MEDIA USED IN CELL CULTURE.docxSUMMARY OF MEDIA USED IN CELL CULTURE.docx
SUMMARY OF MEDIA USED IN CELL CULTURE.docx
PHARMA IQ EDUCATION
 
LABORATORY SETUP OF A CELL CULTURE LAB
LABORATORY SETUP OF A CELL CULTURE LABLABORATORY SETUP OF A CELL CULTURE LAB
LABORATORY SETUP OF A CELL CULTURE LAB
PHARMA IQ EDUCATION
 
Introduction to cell culture- concepts of cell culture part-1
Introduction to cell culture- concepts of cell culture part-1Introduction to cell culture- concepts of cell culture part-1
Introduction to cell culture- concepts of cell culture part-1
PHARMA IQ EDUCATION
 
PORTAL HYPERTENSION
PORTAL HYPERTENSIONPORTAL HYPERTENSION
PORTAL HYPERTENSION
PHARMA IQ EDUCATION
 
Regulatory Affairs Interview Questions .pdf
Regulatory Affairs Interview Questions .pdfRegulatory Affairs Interview Questions .pdf
Regulatory Affairs Interview Questions .pdf
PHARMA IQ EDUCATION
 
Hypersensitivity reactions
Hypersensitivity reactions Hypersensitivity reactions
Hypersensitivity reactions
PHARMA IQ EDUCATION
 
EXPERIMENTAL MODELS OF LIVER DAMAGE
EXPERIMENTAL MODELS OF LIVER DAMAGEEXPERIMENTAL MODELS OF LIVER DAMAGE
EXPERIMENTAL MODELS OF LIVER DAMAGE
PHARMA IQ EDUCATION
 
ANTIOXIDANTS FOR HEPATIC CIRRHOSIS.pptx
ANTIOXIDANTS FOR HEPATIC CIRRHOSIS.pptxANTIOXIDANTS FOR HEPATIC CIRRHOSIS.pptx
ANTIOXIDANTS FOR HEPATIC CIRRHOSIS.pptx
PHARMA IQ EDUCATION
 
INFLAMMATORY BOWEL DISEASES
INFLAMMATORY BOWEL DISEASESINFLAMMATORY BOWEL DISEASES
INFLAMMATORY BOWEL DISEASES
PHARMA IQ EDUCATION
 
GENE AND CELL THERAPY FOR CVDs
GENE AND CELL THERAPY FOR CVDs GENE AND CELL THERAPY FOR CVDs
GENE AND CELL THERAPY FOR CVDs
PHARMA IQ EDUCATION
 
GENE THERPY & CELL BASED THERAPY FOR CNS DISORDERS
GENE THERPY & CELL BASED THERAPY FOR CNS DISORDERSGENE THERPY & CELL BASED THERAPY FOR CNS DISORDERS
GENE THERPY & CELL BASED THERAPY FOR CNS DISORDERS
PHARMA IQ EDUCATION
 

More from PHARMA IQ EDUCATION (20)

Immunopharmacology & immunology Unit-1 (PC-660)
Immunopharmacology & immunology Unit-1  (PC-660)Immunopharmacology & immunology Unit-1  (PC-660)
Immunopharmacology & immunology Unit-1 (PC-660)
 
Pain Models-Screening Assay in vitro & in vivo
Pain Models-Screening Assay in vitro & in vivoPain Models-Screening Assay in vitro & in vivo
Pain Models-Screening Assay in vitro & in vivo
 
Caco-2 cell permeability assay for drug absorption
Caco-2 cell permeability assay for drug absorptionCaco-2 cell permeability assay for drug absorption
Caco-2 cell permeability assay for drug absorption
 
Acute Poisoning and it's management.pptx
Acute Poisoning and it's management.pptxAcute Poisoning and it's management.pptx
Acute Poisoning and it's management.pptx
 
BIOTRANSFORMATION UNIT -1 DRUG METABOLISM (1).pptx
BIOTRANSFORMATION UNIT -1 DRUG METABOLISM (1).pptxBIOTRANSFORMATION UNIT -1 DRUG METABOLISM (1).pptx
BIOTRANSFORMATION UNIT -1 DRUG METABOLISM (1).pptx
 
Significance of DMPK in drug discovery.pptx
Significance of DMPK in drug discovery.pptxSignificance of DMPK in drug discovery.pptx
Significance of DMPK in drug discovery.pptx
 
CONTAMINANTS IN CELL CULTURE & PRECAUTIONS.pptx
CONTAMINANTS IN CELL CULTURE & PRECAUTIONS.pptxCONTAMINANTS IN CELL CULTURE & PRECAUTIONS.pptx
CONTAMINANTS IN CELL CULTURE & PRECAUTIONS.pptx
 
UNIT- 10 ANIMAL CELL CULTURE TECHNIQUES (3).pptx
UNIT- 10 ANIMAL CELL CULTURE TECHNIQUES (3).pptxUNIT- 10 ANIMAL CELL CULTURE TECHNIQUES (3).pptx
UNIT- 10 ANIMAL CELL CULTURE TECHNIQUES (3).pptx
 
SUMMARY OF MEDIA USED IN CELL CULTURE.docx
SUMMARY OF MEDIA USED IN CELL CULTURE.docxSUMMARY OF MEDIA USED IN CELL CULTURE.docx
SUMMARY OF MEDIA USED IN CELL CULTURE.docx
 
SUMMARY OF MEDIA USED IN CELL CULTURE.docx
SUMMARY OF MEDIA USED IN CELL CULTURE.docxSUMMARY OF MEDIA USED IN CELL CULTURE.docx
SUMMARY OF MEDIA USED IN CELL CULTURE.docx
 
LABORATORY SETUP OF A CELL CULTURE LAB
LABORATORY SETUP OF A CELL CULTURE LABLABORATORY SETUP OF A CELL CULTURE LAB
LABORATORY SETUP OF A CELL CULTURE LAB
 
Introduction to cell culture- concepts of cell culture part-1
Introduction to cell culture- concepts of cell culture part-1Introduction to cell culture- concepts of cell culture part-1
Introduction to cell culture- concepts of cell culture part-1
 
PORTAL HYPERTENSION
PORTAL HYPERTENSIONPORTAL HYPERTENSION
PORTAL HYPERTENSION
 
Regulatory Affairs Interview Questions .pdf
Regulatory Affairs Interview Questions .pdfRegulatory Affairs Interview Questions .pdf
Regulatory Affairs Interview Questions .pdf
 
Hypersensitivity reactions
Hypersensitivity reactions Hypersensitivity reactions
Hypersensitivity reactions
 
EXPERIMENTAL MODELS OF LIVER DAMAGE
EXPERIMENTAL MODELS OF LIVER DAMAGEEXPERIMENTAL MODELS OF LIVER DAMAGE
EXPERIMENTAL MODELS OF LIVER DAMAGE
 
ANTIOXIDANTS FOR HEPATIC CIRRHOSIS.pptx
ANTIOXIDANTS FOR HEPATIC CIRRHOSIS.pptxANTIOXIDANTS FOR HEPATIC CIRRHOSIS.pptx
ANTIOXIDANTS FOR HEPATIC CIRRHOSIS.pptx
 
INFLAMMATORY BOWEL DISEASES
INFLAMMATORY BOWEL DISEASESINFLAMMATORY BOWEL DISEASES
INFLAMMATORY BOWEL DISEASES
 
GENE AND CELL THERAPY FOR CVDs
GENE AND CELL THERAPY FOR CVDs GENE AND CELL THERAPY FOR CVDs
GENE AND CELL THERAPY FOR CVDs
 
GENE THERPY & CELL BASED THERAPY FOR CNS DISORDERS
GENE THERPY & CELL BASED THERAPY FOR CNS DISORDERSGENE THERPY & CELL BASED THERAPY FOR CNS DISORDERS
GENE THERPY & CELL BASED THERAPY FOR CNS DISORDERS
 

Recently uploaded

Know the difference between Endodontics and Orthodontics.
Know the difference between Endodontics and Orthodontics.Know the difference between Endodontics and Orthodontics.
Know the difference between Endodontics and Orthodontics.
Gokuldas Hospital
 
June 2024 Oncology Cartoons By Dr Kanhu Charan Patro
June 2024 Oncology Cartoons By Dr Kanhu Charan PatroJune 2024 Oncology Cartoons By Dr Kanhu Charan Patro
June 2024 Oncology Cartoons By Dr Kanhu Charan Patro
Kanhu Charan
 
What are the different types of Dental implants.
What are the different types of Dental implants.What are the different types of Dental implants.
What are the different types of Dental implants.
Gokuldas Hospital
 
Recent advances on Cervical cancer .pptx
Recent advances on Cervical cancer .pptxRecent advances on Cervical cancer .pptx
Recent advances on Cervical cancer .pptx
DrGirishJHoogar
 
Travel Clinic Cardiff: Health Advice for International Travelers
Travel Clinic Cardiff: Health Advice for International TravelersTravel Clinic Cardiff: Health Advice for International Travelers
Travel Clinic Cardiff: Health Advice for International Travelers
NX Healthcare
 
13. PROM premature rupture of membranes
13.  PROM premature rupture of membranes13.  PROM premature rupture of membranes
13. PROM premature rupture of membranes
TigistuMelak
 
Acute Gout Care & Urate Lowering Therapy .pdf
Acute Gout Care & Urate Lowering Therapy .pdfAcute Gout Care & Urate Lowering Therapy .pdf
Acute Gout Care & Urate Lowering Therapy .pdf
Jim Jacob Roy
 
Hemodialysis: Chapter 5, Dialyzers Overview - Dr.Gawad
Hemodialysis: Chapter 5, Dialyzers Overview - Dr.GawadHemodialysis: Chapter 5, Dialyzers Overview - Dr.Gawad
Hemodialysis: Chapter 5, Dialyzers Overview - Dr.Gawad
NephroTube - Dr.Gawad
 
Ageing, the Elderly, Gerontology and Public Health
Ageing, the Elderly, Gerontology and Public HealthAgeing, the Elderly, Gerontology and Public Health
Ageing, the Elderly, Gerontology and Public Health
phuakl
 
Top Travel Vaccinations in Manchester
Top Travel Vaccinations in ManchesterTop Travel Vaccinations in Manchester
Top Travel Vaccinations in Manchester
NX Healthcare
 
Osvaldo Bernardo Muchanga-GASTROINTESTINAL INFECTIONS AND GASTRITIS-2024.pdf
Osvaldo Bernardo Muchanga-GASTROINTESTINAL INFECTIONS AND GASTRITIS-2024.pdfOsvaldo Bernardo Muchanga-GASTROINTESTINAL INFECTIONS AND GASTRITIS-2024.pdf
Osvaldo Bernardo Muchanga-GASTROINTESTINAL INFECTIONS AND GASTRITIS-2024.pdf
Osvaldo Bernardo Muchanga
 
Pollen and Fungal allergy: aeroallergy.pdf
Pollen and Fungal allergy: aeroallergy.pdfPollen and Fungal allergy: aeroallergy.pdf
Pollen and Fungal allergy: aeroallergy.pdf
Chulalongkorn Allergy and Clinical Immunology Research Group
 
How to choose the best dermatologists in Indore.
How to choose the best dermatologists in Indore.How to choose the best dermatologists in Indore.
How to choose the best dermatologists in Indore.
Gokuldas Hospital
 
biomechanics of running. Dr.dhwani.pptx
biomechanics of running.   Dr.dhwani.pptxbiomechanics of running.   Dr.dhwani.pptx
biomechanics of running. Dr.dhwani.pptx
Dr. Dhwani kawedia
 
Local anesthetics 2024/ Medicinal Chemistry pdf
Local anesthetics 2024/ Medicinal Chemistry pdfLocal anesthetics 2024/ Medicinal Chemistry pdf
Local anesthetics 2024/ Medicinal Chemistry pdf
NarminHamaaminHussen
 
Histololgy of Female Reproductive System.pptx
Histololgy of Female Reproductive System.pptxHistololgy of Female Reproductive System.pptx
Histololgy of Female Reproductive System.pptx
AyeshaZaid1
 
Tele Optometry (kunj'sppt) / Basics of tele optometry.
Tele Optometry (kunj'sppt) / Basics of tele optometry.Tele Optometry (kunj'sppt) / Basics of tele optometry.
Tele Optometry (kunj'sppt) / Basics of tele optometry.
Kunj Vihari
 
Artificial Intelligence Symposium (THAIS)
Artificial Intelligence Symposium (THAIS)Artificial Intelligence Symposium (THAIS)
Artificial Intelligence Symposium (THAIS)
Josep Vidal-Alaball
 
NARCOTICS- POLICY AND PROCEDURES FOR ITS USE
NARCOTICS- POLICY AND PROCEDURES FOR ITS USENARCOTICS- POLICY AND PROCEDURES FOR ITS USE
NARCOTICS- POLICY AND PROCEDURES FOR ITS USE
Dr. Ahana Haroon
 
Physical demands in sports - WCSPT Oslo 2024
Physical demands in sports - WCSPT Oslo 2024Physical demands in sports - WCSPT Oslo 2024
Physical demands in sports - WCSPT Oslo 2024
Torstein Dalen-Lorentsen
 

Recently uploaded (20)

Know the difference between Endodontics and Orthodontics.
Know the difference between Endodontics and Orthodontics.Know the difference between Endodontics and Orthodontics.
Know the difference between Endodontics and Orthodontics.
 
June 2024 Oncology Cartoons By Dr Kanhu Charan Patro
June 2024 Oncology Cartoons By Dr Kanhu Charan PatroJune 2024 Oncology Cartoons By Dr Kanhu Charan Patro
June 2024 Oncology Cartoons By Dr Kanhu Charan Patro
 
What are the different types of Dental implants.
What are the different types of Dental implants.What are the different types of Dental implants.
What are the different types of Dental implants.
 
Recent advances on Cervical cancer .pptx
Recent advances on Cervical cancer .pptxRecent advances on Cervical cancer .pptx
Recent advances on Cervical cancer .pptx
 
Travel Clinic Cardiff: Health Advice for International Travelers
Travel Clinic Cardiff: Health Advice for International TravelersTravel Clinic Cardiff: Health Advice for International Travelers
Travel Clinic Cardiff: Health Advice for International Travelers
 
13. PROM premature rupture of membranes
13.  PROM premature rupture of membranes13.  PROM premature rupture of membranes
13. PROM premature rupture of membranes
 
Acute Gout Care & Urate Lowering Therapy .pdf
Acute Gout Care & Urate Lowering Therapy .pdfAcute Gout Care & Urate Lowering Therapy .pdf
Acute Gout Care & Urate Lowering Therapy .pdf
 
Hemodialysis: Chapter 5, Dialyzers Overview - Dr.Gawad
Hemodialysis: Chapter 5, Dialyzers Overview - Dr.GawadHemodialysis: Chapter 5, Dialyzers Overview - Dr.Gawad
Hemodialysis: Chapter 5, Dialyzers Overview - Dr.Gawad
 
Ageing, the Elderly, Gerontology and Public Health
Ageing, the Elderly, Gerontology and Public HealthAgeing, the Elderly, Gerontology and Public Health
Ageing, the Elderly, Gerontology and Public Health
 
Top Travel Vaccinations in Manchester
Top Travel Vaccinations in ManchesterTop Travel Vaccinations in Manchester
Top Travel Vaccinations in Manchester
 
Osvaldo Bernardo Muchanga-GASTROINTESTINAL INFECTIONS AND GASTRITIS-2024.pdf
Osvaldo Bernardo Muchanga-GASTROINTESTINAL INFECTIONS AND GASTRITIS-2024.pdfOsvaldo Bernardo Muchanga-GASTROINTESTINAL INFECTIONS AND GASTRITIS-2024.pdf
Osvaldo Bernardo Muchanga-GASTROINTESTINAL INFECTIONS AND GASTRITIS-2024.pdf
 
Pollen and Fungal allergy: aeroallergy.pdf
Pollen and Fungal allergy: aeroallergy.pdfPollen and Fungal allergy: aeroallergy.pdf
Pollen and Fungal allergy: aeroallergy.pdf
 
How to choose the best dermatologists in Indore.
How to choose the best dermatologists in Indore.How to choose the best dermatologists in Indore.
How to choose the best dermatologists in Indore.
 
biomechanics of running. Dr.dhwani.pptx
biomechanics of running.   Dr.dhwani.pptxbiomechanics of running.   Dr.dhwani.pptx
biomechanics of running. Dr.dhwani.pptx
 
Local anesthetics 2024/ Medicinal Chemistry pdf
Local anesthetics 2024/ Medicinal Chemistry pdfLocal anesthetics 2024/ Medicinal Chemistry pdf
Local anesthetics 2024/ Medicinal Chemistry pdf
 
Histololgy of Female Reproductive System.pptx
Histololgy of Female Reproductive System.pptxHistololgy of Female Reproductive System.pptx
Histololgy of Female Reproductive System.pptx
 
Tele Optometry (kunj'sppt) / Basics of tele optometry.
Tele Optometry (kunj'sppt) / Basics of tele optometry.Tele Optometry (kunj'sppt) / Basics of tele optometry.
Tele Optometry (kunj'sppt) / Basics of tele optometry.
 
Artificial Intelligence Symposium (THAIS)
Artificial Intelligence Symposium (THAIS)Artificial Intelligence Symposium (THAIS)
Artificial Intelligence Symposium (THAIS)
 
NARCOTICS- POLICY AND PROCEDURES FOR ITS USE
NARCOTICS- POLICY AND PROCEDURES FOR ITS USENARCOTICS- POLICY AND PROCEDURES FOR ITS USE
NARCOTICS- POLICY AND PROCEDURES FOR ITS USE
 
Physical demands in sports - WCSPT Oslo 2024
Physical demands in sports - WCSPT Oslo 2024Physical demands in sports - WCSPT Oslo 2024
Physical demands in sports - WCSPT Oslo 2024
 

Histopathological staining techniques used in liver diseases

  • 1. Hematoxylin & Eosin Stain Part-1 of Microscopic tissue fixation techniques used in pharmacological studies Priyansha Singh B. Pharm, M.S. (Pharm.)- Pharmacology & Toxicology
  • 2. The sections of tissues as they are prepared are colourless & different components cannot be identified. Staining makes them differently coloured as various dyes have affinities towards specific components of tissues, making the identification & morphological studies possible. Hematoxylin & Eosin- most frequently used stain in histology
  • 3. INTRODUCTION • Hematoxylin and eosin staining technique functions to recognize different types of tissues and their morphological changes, especially in cancer diagnosis. • It is mainly used as a gold standard stain in medical diagnosis • Hematoxylin has a deep blue-purple color and stains nucleic acids. • Eosin is pink and stains proteins non-specifically. • In a typical tissue, nuclei are stained blue, whereas the cytoplasm and extracellular matrix have varying degrees of pink staining. • Hematoxylin and eosin are both dyes, and naturally, dyes have a high affinity for tissues, depending on the Acidity and/or alkalinity of the dyes.
  • 4. OBJECTIVE To demonstrate the morphologies of tissues and cells
  • 5. PRINCIPLE • Haematoxylin is a dye which when combined with a mordant like potash alum starts to behave like a basic/ cationic dye. The staining of nucleus is blue due to binding of dye metal complex with the nuclear material. • Eosin dye is acidic dye hence it as a negative charge (eosinophilic). Therefore it stains the basic protein structures of a cell & extracellular matrix (acidophils or eosinophils), giving them a red or pink color, for example, the cytoplasm is positively charged, and therefore it will take up the eosin dye, and appear pink.
  • 6.
  • 7. Main types of staining seen on H&E stain
  • 9. PROCEDURE Materials Required • Xylene I & II- for deparaffinization & clearance • 70%, 80%, 100% alcohol- for rehydration & dehydration after counter staining • Haematoxylin- for nuclear staining • 1% acid alcohol- for differentiation • Li2CO3- for bluing • Eosin- counter staining • DPX- mounting
  • 11. • Deparaffinization- to remove wax embedded in the tissue. • Descending grades of alcohol- After thorough de-waxing, the slide is passed through several changes of alcohol to remove the xylene, then thoroughly rinsed in water. The section is now hydrated so that aqueous reagents will readily penetrate the cells and tissue elements. • Haematoxylin- for nuclear staining • Differentiation- to remove non-specific background staining and to improve contrast. • Bluing- changes the color of hematoxylin from red to blue by chemically altering the dye with a slightly basic solution. The bluing solution's alkaline pH stabilizes the stain and makes it insoluble in water and alcohol. The change in pH is what causes the color change. • Counterstaining- A counterstain is a stain with color contrasting to the principal stain, making the stained structure easily visible using a microscope. • Ascending grades of alcohol- for dehydration. To remove water from the tissue so that paraffin wax can completely penetrate it. Since wax is not water soluble, but is soluble in xylene, the water in the tissue must be replaced with xylene. • Clearance by Xylene I & II- to remove dehydrating agents and prepare tissues for impregnation with the embedding agent. Clearing makes the tissue transparent, or clear, making the slides easier to read. • DPX mounting- to increase the refractive index and improve image quality in microscope
  • 12. RESULTS • Nuclei are stained blue, cytoplasm & ECM- varied degrees of pink stain • Well fixed cells/ tissues show considerable intracellular details • Nuclei stains with eosin • If polyribosomes are present then cytoplasm will have a distinct blue cast
  • 13. Masson’s Trichrome Staining Part-2 of Microscopic tissue fixation techniques used in pharmacological studies Priyansha Singh B. Pharm, M.S. (Pharm.)- Pharmacology & Toxicology
  • 14. Definition • Masson’s trichrome is a differential staining technique for selective staining of collagen fibres, fibrin, muscles & RBCs. • Trichrome stain- Wiegert's haematoxylin (nucleus) - Biebrich scarlet (muscle, cytoplasm & nucleus) - Aniline/ methylene blue/ light green (collagen)
  • 15. Objectives of Masson’s Trichrome staining • To stain the collage fibers. • To stain collagen. • To stain keratin. • To stain fibrin. • To stain muscle fibers. • To differentiate between collagen & smooth muscle in tumors, increase in collagen in diseases like cirrhosis. Routine stain in kidney fibrosis & liver cirrhosis biopsies.
  • 16. Principal behind the staining • As the name implies, 3 dyes are employed selectively staining muscle, collagen fibers, fibrin & erythrocytes. • The general rule in trichrome staining is that the less porous tissues are coloured by the smallest size dye molecule. • Wherever a dye of large molecular size is able to penetrate but the dye of smaller size passes off the larger pore size. • The tissue is first stained with the acid dye (Biebrich Scarlet red- Acid Fuchsin) binding with acidophilic tissue components. • Further treatment with the phospho acids, the less permeable components retain red stain and the red colour comes out of the collagen pores • Collagen gets bind with aniline blue.
  • 17.
  • 18. Fix the sample using Bouin’s solution- mordant (microwave for 1 min) stand for 15 mins Add Wiegert's Haematoxylin to stain nucleus (10 mins) Add Biebrich scarlet red stain (5 mins) to stain cytoplasm, keratin, muscle & RBCs Add Phosphotungstic/ phosphomolybdic acid for decolorization (10 mins). Add aniline blue/ methylene blue/ light green + 1% acetic acid to stain collagen Wash (5 mins) Wash & rinse Rinse Procedure of staining
  • 19. Results Nuclei black Cytoplasm, Muscle, Erythrocytes- red Collagen- blue ADVANTAGES 1. To measure & differentiate collagen deposition as compared to HE staining 2. Produces clear images of collagen compared to other stains 3. More accurate than other staining techniques DISADVANTAGES 1. Preparation of solutions to be used within 24 hrs 2. Corrosive stains
  • 20. Sirius red staining Part-3 of Microscopic tissue fixation techniques used in pharmacological studies Priyansha Singh B. Pharm, M.S. (Pharm.)- Pharmacology & Toxicology