The document provides details about a course on special histological techniques including immune histochemistry and molecular pathology. The course aims to teach advanced histological techniques like enzyme histochemistry, immune histochemistry, and molecular pathology techniques. It includes 3 units that cover special staining techniques, principles of enzyme histochemistry, immune histochemistry techniques, and molecular pathology techniques like in situ hybridization, polymerase chain reaction, and flow cytometry. The syllabus and course objectives aim to develop skills in these advanced techniques and their applications in pathology.

1. Department of Medical Laboratory Technology, UIAHS.
Course Code: 21MSH-716 Course Name: Special Histological techniques including Immune Histochemistry and molecular pathology
Enzyme Histochemistry
Name of the Faculty: Mr. Attuluri. Vamsi Kumar E13404 Program Name: M.Sc. MLT

2. Program Structure MSc MLT 2021
Semester-I
Course
Code
Course Name L T P C CH
per
week
Course
Category
Theory
Assessment
Practical
Assessment
Total
IA MTA ETA CA MTA ETA
21MSH-
611
Cellular
Histopathology
3 0 3 4.5
6
PC
20 20 60 40 20 40 200
21MSH-
612
Applied Clinical
Biochemistry
3 0 3 4.5
6
PC
20 20 60 40 20 40 200
21MSH-
613
Diagnostic
instrumentation
3 0 3 4.5
6
PC
20 20 60 40 20 40 200
21MSH-
614
Clinical
Immunopatholog
y
3 0 3 4.5
6
PC
20 20 60 40 20 40 200
Total Credits 18 24
Cumulative Credits 18 24

3. Program Structure MSc MLT 2021
Semester-II
Course
Code
Course Name L T P C CH
per
week
Course
Category
Theory Assessment Practical
Assessment
Total
IA MTA ETA CA MTA ETA
21MSH-651
Diagnostic
Enzymology 3 0 3 4.5
6
PC
20 20 60 40 20 40 200
21MSH-652
Clinical
Immunology 3 0 3 4.5
6
PC
20 20 60 40 20 40 200
21MSH-653
Molecular biology
&Genetics 3 0 3 4.5
6
PC
20 20 60 40 20 40 200
21MST-654
Advance
Hematology
3 0 3 4.5
6
PC
20 20 60 40 20 40 200
21MST-659
Domain Aptitude
3 0 0 2
2
PC
20 20 60 0 0 0 100
Total Credits 20 26
Cumulative Credits 20 26

4. Program Structure MSc MLT 2021
Semester-III
Course Code Course Name L T P C CH
per
week
Course
Categor
y
Theory Assessment Practical
Assessment
Total
IA MTA ETA CA MTA ETA
21MSA-701
Blood Banking
3 0 0 3
3
SE
20 20 60 0 0 0 100
21MSA-702
Hematology
3 0 3 4.5
6
SE
20 20 60 20 20 60 200
21MSB-701
Basic and Advanced
Histological Techniques 3 0 3 4.5
6
SE
20 20 60 20 20 60
21MSH-716 Special Histological
Techniques Including
Immune Histochemistry
and molecular
pathology
3 0 3 3
3
SE
20 20 60 0 0 0 100
21MSH-701
Virology and Myology
3 0 3 4.5
6
SE
20 20 60 20 20 60

5. 21MSH-702
Medical Parasitology
3 0 3 4.5
6
SE
20 20 60 20 20 60 200
21MSH-701
Advance Clinical Bio-
Chemistry 3 0 3 4.5
6
SE
20 20 60 20 20 60 200
21MSH-702
Advance Techniques Used In
Clinical Bio-Chemistry 3 0 3 4.5
6
SE
20 20 60 20 20 60 200
21MST-703
Research Methodology and
Biostatics 3 0 3
3
PC
20 20 60 0 0 0 100
21MSR-704
Dissertation-I
0 0 10
10
PC
0 0 0 40 20 40 100
21MSY-710 Good Clinical Lab Practices
1 0 0 1
1
MNG
20 20 60 0 0 0 100
21MSY-711 Quality Management
System 1 0 0 1
1 MNG 20 20 60 0 0 0 100
21MSY-712 Intricacies of NABL
1 0 0 1
1 MNG 20 20 60 0 0 0 100
Total Credits 21.5 25
Cumulative Credits 21.5 25
Program Structure MSc MLT 2021

6. Course Objectives
This course aims to learn the basic concepts and to get high standard knowledge of histo
related to skin, respiratory, digestive, excretory, and reproductive organ systems
Students will learn about the histo pathologies of various organs of human body
This course aims to learn the basic concepts and to get high standard knowledge of histo
related to skin, respiratory, digestive, excretory, and reproductive organ systems

7. Course outcomes
CO1 At the end of the course the candidate should be able to define the structural features of
respiratory, digestive, excretory, and reproductive organ systems.
CO2 To develop an understanding of the anatomical pathologies of various organs of human body
CO3 The candidate should be capable of understanding cytology and histopathology of various organ
systems.Student should be capable of providing optimal pathology services in any medical set up
and develop research aptitude at PG level in the subject.
CO4 At the end of the course the candidate should be able to define the structural features of
respiratory, digestive, excretory, and reproductive organ systems.

8. Syllabus overview
Unit.1 Special Techniques For Histochemical Studies, Principles & Advanced
Special Staining Techniques & enzyme Histo chemistry
Contact Hours:15
Chapter 1 Freeze drying & vapor fixation (Special techniques),
Principles of following: hemoglobin, Argentaffin cell granules, Axons, Purkinje cells, Adernal medulla, Peneth
Cells, Astrocytes, Glial fibres, Oligodendrocytes, Neurofibrills, Nerve ending and microglia
Chapter 2 Enzyme Histochemistry: Preparation of post – fixed and prefixed cryostat sections, Demostration of following
enzymes:
i) Phosphatase – Acid Phase, alkaline phase & ATP
ii) Dehydrogenases
iii) Oxidases
iv) Esterases
v) Tyrosinase
Experiment 1 Demonstration of different fixatives used in Histo
Experiment 2 Demonstration of different Microtome used in Histo
Experiment 3 To demonstrate the activity of enzym
Experiment 4 Demonstration of following enzymes activity in a Ti

9. Syllabus overview
Unit-2 Immunohistochemistry Contact Hours:15
Chapter 1 i) Principle of direct, indirect and other special techniques
ii) FIC and FITC techniques
i) Techniques of unmasking of antigen sites.
Chapter 2 i) Enhancement and amplification of binding sites
ii) Double or multiple labeling techniques
iii) Preparation of conjugates.
Chapter 3
i) Blocking of endogenous enzyme and background
ii) Methods foe frozen and paraffin sections
iii) Controls, internal & external quality assessment.
Experiment 5 Demonstration of Laboratory method that uses antib
Experiment 6 Demonstrate the FIC and FITC techniques
Experiment 7 Demonstration of the technique used to separate DN

10. Syllabus overview
Unit-3 Molecular Pathology Contact Hours:15
Chapter 1 Molecular Pathology:
In Situ-hybrdisation : Principle, techniques and their application, Probes and Application
in fixed tissue sample.
Chapter 2 Polymerase Chain Reaction: Principle and types with their utilization of Polymerase Chain
Reactions
Electrophoresis : Principle, types and their applications.
Chapter 3 Image Analysis: Overview, technique, specimen preparation and analysis.
Flow Cytometry: Principle, Technique, Specimen collection and preparation and their
application
Experiment 8 Demonstration of technique for rapidly producing
Experiment 9 Demonstration about the technique that is used for
Experiment 10 Demonstrate the Flow cytometer technique used to

11. NOTES
Basics about enzymes

15. Properties of Enzymes & Factors affecting enzyme activity

19. Notes
Enzyme Histochemistry

20. Introduction

21. Significance of histochemistry

24. General Principles

25. Factors influencing the reaction

26. Factors influencing in enzyme histochemistry

27. Factors influencing in enzyme histochemistry

28. Factors influencing in enzyme histochemistry

29. Controls & Fixation

30. Fixation

31. Sections preparation

32. Principles of Enzyme histochemistry & Types of
histochemical reactions

33. Types of histochemical reactions

34. Principles of Enzyme histochemistry & Types of
histochemical reactions

35. Types of histochemical reactions

36. Principles of Enzyme histochemistry & Types of
histochemical reactions

37. Types of histochemical reactions

38. Principles of Enzyme histochemistry & Types of
histochemical reactions

39. Types of histochemical reactions

40. Demonstration of Alkaline Phosphatase

41. Clinical Significance of Alkaline Phosphatase (ALP)

42. Clinical Significance of Alkaline Phosphatase
(ALP)

43. 6.2.1 Alkaline phosphatase (AKP)
AKP is a hydrolytic enzyme. It hydrolysis various phosphate esters of alcohol and phenol (such as sodium
β-glycerophosphate and phosphoric acid naphthalene ester) in alkaline condition (pH 9.0–9.6). The
hydrolytic activity could be activated by Mg2+, Mn2+, Zn2+ and Co2+ and easily inhibited by cysteine, cyanide
and arsenic acid salt. In most tissues, the AKP could be selectively inhibited by tetramisole.
Demonstration of Alkaline Phosphatase (ALP)

44. Demonstration of Alkaline Phosphatase (ALP)
AKP
AKP
AKP
AKP
AKP
AKP
AKP
AKP
AKP
AKP
Alpha
Napthol
Phosphate
H20 –
pH 9.2
Napth
onol
Phosphate
Diazonium
Salt
Azopigment

46. PROCEDURE
1. Fresh tissue is fixed with 10% neutral formaldehyde for 12–24 hours or with 95%
acetone-liquid alcohol for 4–12 hours (change the fixative for two times, room
temperature)
2. Dehydrate the sections with pure alcohol, clear with xylene, embed with paraffin
and then section.
3. Dewax and rinse the sections with water for three times.

47. Syllabus overview
Unit.1 Special Techniques For Histochemical Studies, Principles & Advanced
Special Staining Techniques & enzyme Histo chemistry
Contact Hours:15
Chapter 1 Freeze drying & vapor fixation (Special techniques),
Principles of following: hemoglobin, Argentaffin cell granules, Axons, Purkinje cells, Adernal medulla, Peneth
Cells, Astrocytes, Glial fibres, Oligodendrocytes, Neurofibrills, Nerve ending and microglia
Chapter 2 Enzyme Histochemistry: Preparation of post – fixed and prefixed cryostat sections, Demostration of following
enzymes:
i) Phosphatase – Acid Phase, alkaline phase & ATP
ii) Dehydrogenases
iii) Oxidases
iv) Esterases
v) Tyrosinase
Experiment 1 Demonstration of different fixatives used in Histo
Experiment 2 Demonstration of different Microtome used in Histo
Experiment 3 To demonstrate the activity of enzym
Experiment 4 Demonstration of following enzymes activity in a Ti

51. Syllabus overview
Unit.1 Special Techniques For Histochemical Studies, Principles & Advanced
Special Staining Techniques & enzyme Histo chemistry
Contact Hours:15
Chapter 1 Freeze drying & vapor fixation (Special techniques),
Principles of following: hemoglobin, Argentaffin cell granules, Axons, Purkinje cells, Adernal medulla, Peneth
Cells, Astrocytes, Glial fibres, Oligodendrocytes, Neurofibrills, Nerve ending and microglia
Chapter 2 Enzyme Histochemistry: Preparation of post – fixed and prefixed cryostat sections, Demostration of following
enzymes:
i) Phosphatase – Acid Phase, alkaline phase & ATP
ii) Dehydrogenases
iii) Oxidases
iv) Esterases
v) Tyrosinase
Experiment 1 Demonstration of different fixatives used in Histo
Experiment 2 Demonstration of different Microtome used in Histo
Experiment 3 To demonstrate the activity of enzym
Experiment 4 Demonstration of following enzymes activity in a Ti

52. Histology of nervous system

58. FUNCTIONAL UNIT OF NERVOUS SYSTEM
IS

80. Reference
1. https://pt.slideshare.net/HusseinAltameemi2/cardiac-biomarkers-i
2. https://www.pdfdrive.com/macaca-mulatta-enzyme-
histochemistry-of-the-nervous-system-d165642716.html
3. https://www.pdfdrive.com/a-short-history-of-histochemistry-in-
denmark-e60339299.html
4. https://www.pdfdrive.com/enzyme-histochemistry-a-laboratory-
manual-d183701064.html
5. https://www.slideshare.net/KrishnenduSinha1/histochemistry-
235325874