This course, "Applied Histopathology-II", expands upon the basic skills and knowledge introduced in the prerequisite Histopathology course. It provides a comprehensive understanding of various staining procedures, cryostat sectioning, enzyme histochemistry, and electron microscopic procedures in histopathology. The course focuses on the detection, demonstration, and identification of different substances like connective tissue elements, proteins, nucleic acids, carbohydrates, lipids, etc. It also incorporates practical experiments to give students hands-on experience in these techniques. Created by: Mr. Attuluri Vamsi Kumar, Assistant Professor, Department of MLT, UIAHS, Chandigarh University, Mohali, Punjab. For more details website: https://www.mltmaster.com

1. B.Sc. MLT Syllabus
Attuluri Vamsi Kumar I Assistant professor I Dept of MLT I Ph No: 7416660584 I Website: mltmaster.com
Website: www.mltmaster.com I YouTube: https://www.youtube.com/@vamsiMLT
Program Name: B.Sc. Medical Lab Technology
Syllabus of Applied Histopathology - II
Course Name: Applied Histopathology - II Course Type
Course Coordinator: Attuluri Vamsi Kumar – B.Sc. MLT, M.Sc. MLT, PhD (Perusing)
Designation: Assistant Professor
Department: Department of Medical Lab Technology (MLT)
Program Core
PRE-REQUISITE Applied Histopathology – I Sem: 5 S. No: 36
About Vamsi: I am academician in Medical Laboratory Sciences with a strong desire to improve Outcome
based education (OBE) structured MLT education. I am constantly focusing on building an academic
atmosphere that is set high standards with strong multi blended teaching pedagogy models.
YouTube: https://www.youtube.com/@vamsiMLT
SlideShare: https://www.slideshare.net/VamsiIntellectual
Website: www.mltmaster.com / https://sites.google.com/view/vamsi-intellectual-protfolio/home
LinkedIn: https://www.linkedin.com/in/vamsi-kumar-attuluri-ab8987128/?originalSubdomain=in
Research Gate: https://www.researchgate.net/profile/Attuluri-Kumar
Orcid ID: https://orcid.org/0000-0001-9278-6714
Contact No: +91 7416660584
Mail ID: vamsifmlt@gmail.com
A. COURSE DESCRIPTION
This course, "Applied Histopathology-II", expands upon the basic skills and knowledge
introduced in the prerequisite Histopathology course. It provides a comprehensive
understanding of various staining procedures, cryostat sectioning, enzyme histochemistry, and
electron microscopic procedures in histopathology. The course focuses on the detection,
demonstration, and identification of different substances like connective tissue elements,
proteins, nucleic acids, carbohydrates, lipids, etc. It also incorporates practical experiments to
give students hands-on experience in these techniques.
B. COURSE OBJECTIVES:
1. Define and describe the different staining procedures used in histopathology, including special
staining and metal impregnation techniques.
2. Explain the principles and applications of cryostat sectioning in diagnostic histopathology, and the
importance of enzyme histochemistry.
3. Apply different staining techniques to demonstrate the presence of proteins, nucleic acids,
carbohydrates, lipids, bacteria, fungi, and other elements in tissue sections.
4. Analyze tissue specimens, interpret staining results, and differentiate between different substances
based on their staining characteristics.
5. Evaluate the quality of staining, the integrity of tissue sections, and troubleshoot any issues
encountered during the staining process.
C. COURSE OUTCOMES
CO
No
Statement Performance
Indicator
Level of
Learning
(Highest BT
Level)
Target
Attainment
CO1 Recall and understand the key concepts and
terminologies related to various histopathology
techniques, including special staining procedures,
enzyme histochemistry, cryostat sectioning, and
electron microscopic procedures.
PI 1, PI 6 K1: Knowledge 70%

2. B.Sc. MLT Syllabus
Attuluri Vamsi Kumar I Assistant professor I Dept of MLT I Ph No: 7416660584 I Website: mltmaster.com
Website: www.mltmaster.com I YouTube: https://www.youtube.com/@vamsiMLT
CO2 Interpret the principles and applications of these
histopathological techniques, and explain their
significance in the diagnosis and study of various
diseases.
PI 2.5, PI 3, PI
6
K2:
Comprehension
70%
CO3 Demonstrate proficiency in using these techniques
to prepare, stain, and analyze histological
specimens, and apply these skills to diagnose
pathological conditions.
PI 2.1, PI 2.4,
PI 2.5, PI 5
K3: Application 70%
CO4 Analyze histopathological images, identify
abnormalities, and interpret results based on the
staining characteristics and the morphological
features of the tissues.
PI 2.6, PI 2.7,
PI 2.8
K4: Analysis 70%
CO5 Critically evaluate and troubleshoot
histopathological techniques, ensuring quality
control and accurate diagnostic reporting.
PI 2.9, PI 3, PI
4
K5: Evaluation 70%
CO6 Develop and execute a series of histopathology
procedures, leading to the creation of diagnostic
slides, and communicate these findings effectively
in a professional setting.
PI 1, PI 2.2, PI
2.3, PI 7, PI 8,
PI 9, PI 10
K6: Creation 70%
D. SYLLABUS
Unit-1 Basic Histopathology and Staining Procedures Contact Hours:15
Chapter 1.1 Cryostat Sectioning and Special Staining- Cryostat sectioning, its applications in diagnostic
histopathology.,
Special Staining Procedures for detection of Connective tissue elements, Trichrome staining,
muscle fibers, elastic, reticulin fibers, collagen fibers etc.,
Metachromatic staining suc
Chapter 1.2 Demonstration of Biochemical Substances and Special Treatment- Demonstration and
identification of minerals and pigments, removal of Pigments/artifacts in tissue sections.,
Demonstration of Proteins & nucleic acids.,
Demonstration of Carbohydrates, lipids, fat & fat like substances.,
Tissue requiring special treatment i.e. eye ball, bone marrow, and muscle biopsy, under calcified
or unclarified bones, whole brain, and whole lungs including other large organs.
Experiment 1 /
Case study
To cut frozen section and stain for Haematoxylin and Eosin, Metachromatic stain
Toluidine blue-‗o‘ and Oil Red ‗O‘ staining for the demonstration of fat.
Experiment 2 /
Case study
To prepare Schiff‘s reagent in the lab and do Periodic Acid Schiff‘s (PAS) stain on a paraffin
section.
Experiment 3 /
Case study
To prepare ammonical silver bath in the laboratory and stain paraffin embedded section for the
demonstration of reticulin fibers.
Unit-2 Enzyme Histochemistry and Specialized Techniques Contact Hours:15
Chapter 2.1 Enzyme Histochemistry and Vital Staining - 36
Chapter 2.2 Neuro-pathological Techniques and Museum Techniques- Neuro-pathological techniques.,
Museum techniques.
Experiment 4 /
Case study
To stain a paraffin section for the demonstration of smooth muscle by Van Gieson‘s Stain.
Experiment 5 /
Case study
To perform Masson‘s trichrome stain on a paraffin section for the demonstration of collagen
fiber, muscle fiber and other cell elements.
Experiment 6 /
Case study
To stain the paraffin section for the demonstration of the elastic fibers (EVG).
Experiment 7 /
Case study
To stain Decalcified paraffin embedded section for the presence of calcium salts (Von Kossa‘s
method).
Unit-3 Advanced Histopathology Techniques Contact Hours:15

3. B.Sc. MLT Syllabus
Attuluri Vamsi Kumar I Assistant professor I Dept of MLT I Ph No: 7416660584 I Website: mltmaster.com
Website: www.mltmaster.com I YouTube: https://www.youtube.com/@vamsiMLT
Chapter 3.1 Electron Microscope Techniques- Electron Microscope: working principle and its
components.,
Processing, embedding and ultra-microtomy.
Chapter 3.2 Micrometry and Morphometry- Micrometry and Morphometry.Gram‘s staining
AFB staining (Ziehl Neilson‘s staining) for M. tuberculosis and leprae,
Grocott‘s stain for fungi,
Schmorl‘s reaction for reducing substances (melanin)
Experiment 8 /
Case study
To stain a paraffin section for the following Mucicarmine, Alcian blue.
Experiment 9 /
Case study
To stain a paraffin section for the demonstration of iron (Perl‘s stain).
Experiment 10 /
Case study
To demonstrate the presence of bacteria and fungi in paraffin embedded sections using the
following staining procedures:
Gram‘s staining
AFB staining (Ziehl Neilson‘s staining) for M. tuberculosis and leprae
Grocott‘s stain for fungi
Schmorl‘s reaction fo
Self-study topics for Advance learners: Advanced Molecular Pathology: Investigate the
application of molecular biology techniques in histopathology. This includes genetic testing,
biomarker analysis, and precision medicine., Automated Histopathology Image Analysis: Develop a
better understanding of the role of digital pathology and machine learning in histopathology. This
includes algorithms for detecting and classifying diseases from histopathological images.,
Immunohistochemistry and its applications: Delve deeper into the principles of immunohistochemistry
and its clinical application, including the detection of specific proteins in tissue sections., Advanced
Special Staining Techniques: Explore more complex staining techniques beyond those covered in the
course, such as Immunofluorescence, FISH, and CISH.
E. TEXT BOOKS/REFERENCE BOOKS
TEXT BOOKS
T1 Handbook of Histopathological Techniques by C F A Culling
T2 Medical Lab technology by Lynch
T3 An Introduction to Medical Lab Technology by F J Baker and Silverton
REFERENCE BOOKS
R1 Bancroft's Theory and Practice of Histological Techniques by Suvarna SK, Layton C, and
Bancroft JD
R2 Digital Pathology: Historical Perspectives, Current Concepts & Future Applications by
Keith J. Kaplan and Luigi K.F. Rao
R3 Immunohistochemistry: Basics and Methods by Ulrich Schenck and Torsten Goldmann

4. B.Sc. MLT Syllabus
Attuluri Vamsi Kumar I Assistant professor I Dept of MLT I Ph No: 7416660584 I Website: mltmaster.com
Website: www.mltmaster.com I YouTube: https://www.youtube.com/@vamsiMLT
Code Performance Indicators
PI 1 Demonstrate professional interpersonal, oral, and written communications skills sufficient to
serve the needs of patients and the public including an awareness of how diversity may affect
the communication process.
PI 2 Perform pre-analytical, analytical, and post-analytical processes:
PI 2.1 Demonstrate ability to understand investigation/test requisition.
PI 2.2 Collecting the relevant clinical samples along with complete and accurate documentation with
proper safety measures in relation to sample accountability.
PI 2.3 To transport the samples with precautionary measures to the relevant lab section.
PI 2.4 Demonstrate the ability to prepare clinical sample for processing
PI 2.5 To demonstrate the knowledge of accurate sample processing for the required lab
investigation. Perform routine clinical laboratory tests in clinical chemistry,
haematology/haemostasis, immunology, immunohematology, microbiology, Histopathology,
Cytopathology, body fluid analysis, and laboratory operations.
PI 2.6 Perform mathematical calculations related to all areas of the clinical laboratory
PI 2.7 Ability to record the test results/data.
PI 2.8 To demonstrate the ability to interpret the test reports and its documentation in lab records.
PI 2.9 Demonstrate ability to release the report to the right person in minimum turn-around time
(TAT).
PI 3 Perform problem solving and troubleshooting techniques for laboratory methodologies
Correlate laboratory test results with patient diagnosis and treatment.
PI 4 To follow basic quality assessment protocol of clinical laboratory.
PI 5 Demonstrate routine laboratory techniques sufficient to orient new employees within the
clinical laboratory.
PI 6 Apply basic scientific principles in learning new techniques/procedures; demonstrate
application of principles and methodologies.
PI 7 Utilize computer technology applications to interact with computerized instruments and
laboratory information systems.
PI 8 Demonstrate adequate knowledge of computer software as it applies to document production,
spreadsheets, and presentations.
PI 9 Demonstrate professional behaviour with co-team mates.
PI 10 Demonstrate sensitivity and compassion towards patients.