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Histology

Histology

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Sonalika Mahapatra

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Course Name : Histology

Code(Credit) : CUTM1720 (3-1-0)

 

Course Objectives

  • Understanding the concept of histotechnology;Basic concepts about routine methods of examination of tissues Collection.
  • perform routine laboratory procedures encompassing all major areas of the histology laboratory.
  • accurately and proficiently embed tissue and understand the principles of microtomy.
  • Clinically relevant onchological analysis for deeper understanding of abnormal cell growth at anywhere in human body.
  • The conceptual understanding of the subject provides opportunities for  employability and scopes for higher education.

Course Outcomes

COs  Course Outcomes Mapping COs with POs (High-3, Medium-2, Low- 1)
CO1

Understand the basic concepts of histotechnology, including the examination of tissues, collection and transportation of specimens, and fixation.

PO1(3)
CO2  

Acquire knowledge and skills related to the decalcification process, selection of tissue fixation, and tissue processing and embedding.

PO1(3), PO2(3)
CO3

 

 

 

CO4

 

 

Develop practical skills in staining techniques, including Periodic Acid Schiff (PAS) staining, routine H&E stain, and special stains. Understand the principles and use of impregnation and mounting methods in histotechnology.

 

Develop practical skills in tissue processing, including manual and automated processing, section cutting, mounting of stained smears, attachment of tissue sections to glass slides, and drying of sections in an incubator.

PO2(3) PO7(2),

 

 

 

PO8(2)

Course Syllabus

Module- I ( 8 Hrs)

 

Histotechnology, Care & Maintenance of histotechnology equipements and their parts and Safety measures of laboratory equipment used in histotechnology. Basic concepts about routine methods of examination of tissues, Collection and transportation of specimens for histological examination, fixation: Process, Various types of fixatives used in a routine histopathology laboratory- Simple fixatives, Compound fixatives, Special fixatives for demonstration of various tissue elements.

 

Practice: Care & maintenance of  Histology equipments, Collection & transportation of specimens, Fixation

 

Module- II (Hrs)

 

Decalcification Criteria of a good decalcification agent, Technique of decalcification Followed with selection of tissue fixation, decalcification neutralization of acid and thorough washing. Various types of decalcifying fluids, Processing of various tissues for histological examination, Embedding, Schedule for manual or automatic Tissue processing, Components & principles of various types of a tissue processors.

 

Practice: -Method of Decalcification,  Embedding,  manual or automatic tissue processings schedule.

 

Module- III (10 Hrs)

 

Periodic Acid Schiff Staining, Impregnation and Mountains, Commonly used mountains in histotechnology lab. General Staining Procedures( routine H&E stain, PAP stain and other special stain) for Paraffin Infiltrated and Embedded tissue, To perform & practice the manual & automated Haematoxylin and Eosin staining technique, To perform & practice the Mallory’s Phosphotungstic Acid Haematoxylin (PTAH). Introduction of FNAC and its staining tech, museum technique, post mertum technique.

 

Practice: - Procedure for manual Staining and Automatic Staining Technique, FNAC technique, Museum technique(Hospital Visit), Post mertum technique(Hospital Visit).

 

Module- IV (8 Hrs)

 

Demonstration of instruments used for dissection Use of antiseptics, disinfectants and insecticides in a tissue processing laboratory Reception and labeling of histological specimens Preparation of various fixatives -Helly’s fluid, Zenker’s fluid, Bouin’s fluid, Corney’s fluid, 10% Neutral formalin, Formal saline, Formal acetic acid, Pereyn’s fluid,  prepare 70% alcohol from absolute alcohol. To perform embedding and casting of block.

 

Practice:

Use of antiseptics, disinfectants and insecticides in tissue processing laboratory, Preparation of various Fixatives,  Labeling of Histological specimens, Embedding and Casting of block

 

 

Module-(8 Hrs)

 

Tissue Processor,  Microtomy, Honing and Stropping technique, Use of tissue floating bath, Use of incubator

 

Practice:

Processing of tissue by manual and automated processor. method To demonstrate various part and types of microtome. To learn sharpening of microtome knife (Honing and stropping technique) To perform section cutting, learn mounting of stained smears. To practice attachment of tissue sections to glass slides To learn using tissue floatation bath drying of sections in incubator (370 C),

 

Suggested Readings:

  1. Color text book of histology by Gartner &Hiatt, publisherElsevier
  2. Netter’s essential histology by William Ovalle, publisherElsevier
  3. Histology E-book by Barry Mitchell, publisherElsevier
  4. Textbook of Histology (color atlas) by Krishna Garg, Indira Bahl, Mohini kaul, publisherCBS
  5. Textbook of Histology and a Practical Guide by JP Gunasegaran, PublisherElsevier
  6. Textbook of Medical Laboratory Technology by Praful B Godkar, Publisher Bhalami
  7. Ebook link-https://books.google.co.in/books?id=qWScAQAAQBAJ&printsec=frontcover&source=gbs_ge_summary_r&cad=0#v=onepage&q&f=false
  8. Ebook link-https://books.google.co.in/books?id=MrpEDwAAQBAJ&printsec=frontcover&source=gbs_ge_summary_r&cad=0#v=onepage&q&f=false
  9. Ebook link-https://books.google.co.in/books?id=CERPDwAAQBAJ&printsec=frontcover&source=gbs_ge_summary_r&cad=0#v=onepage&q&f=false

 

All the practice components can be performed by simulation as well as virtual labs. Amrita Vlab (https://www.youtube.com/channel/UC1ouqR_dIY_WPDPSZo0e8Wg).

Online Source: Udemy, Coursera, article library, NCBI,Study.com, You tube, Frontier, Febs, unacademy, academia, Slide share, Labs for life, MedlinePlus.

Session 4

Topic: Basic concepts about routine methods of examination of tissues

Study Materials:

Basic Concept- https://webpath.med.utah.edu/HISTHTML/HISTOTCH/HISTOTCH.html

Session 5

Topic: Collection and transportation of specimens for histological examination

Study Materials:

Discussion- https://www.leicabiosystems.com/knowledge-pathway/steps-to-better-specimen-collection-transport/

Session 7

Topic: Various types of fixatives used in a routine histopathology laboratory

Study Materials: 

Discussion- https://www.leicabiosystems.com/knowledge-pathway/fixation-and-fixatives-4-popular-fixative-solutions/

Simple fixatives & Compound fixatives

Video:https://spark.adobe.com/video/HRWoHRovaoYLq

Slide share- https://www.slideshare.net/hitvet305/fixatives-used-in-histopathology

Session 8

Topic: Special fixatives for demonstration of various tissue elements

Study Materials: 

Discussion- https://www.leicabiosystems.com/knowledge-pathway/fixation-and-fixatives-4-popular-fixative-solutions/

Session 9

Practice session:

Topic: Fixation

Study Materials:

Notes- https://spark.adobe.com/page/sgWX0op7sKdV0/

Session 10

Session 11

Practice Session:

Topic: Decalcification (Criteria of a good decalcification agent)

Study Materials:

Notes- https://spark.adobe.com/page/UVrmsuYtOe1ve/

Slide share- https://www.slideshare.net/Leyoxan/decalcification-61481989?qid=a4ffcf29-4c40-4eae-ba5c-5a637fdadcb7&v=&b=&from_search=4

Session 12

Topic: Schedule for manual or automatic Tissue processing

Study Materials:

Article- https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4774282/

Video:https://spark.adobe.com/video/7tWmV59pm3JTo

Session 13

Topic: Components & principles of various types of a tissue processor

Study Materials:

Notes- https://tissuesampling.weebly.com/processing.html

Session 15

Topic: Embedding

Study Materials:

Notes: https://spark.adobe.com/page/cEME4G1CmY8wD/

Session 18

Topic: Commonly used mountains in histotechnology lab

Study Materials:

Article- http://www.jdrntruhs.org/article.asp?issn=2277-8632;year=2014;volume=3;issue=5;spage=1;epage=8;aulast=Ravikumar

Session 19

Practice session:

Topic: Impregnation and Mountains

Study Materials:

Video- https://www.youtube.com/watch?v=VH1vx8XY1ig

https://www.youtube.com/watch?v=iWCLStcz1PE

Session 20

Practice Session:

Topic: To perform & practice the Haematoxylin and Eosin staining technique

Study Materials:

Video- https://www.youtube.com/watch?v=1tqA_gWUjkM&t=119s

https://www.youtube.com/watch?v=1tqA_gWUjkM

https://www.youtube.com/watch?v=2D0rj0m6dVs&t=10s

Session 21

Practice session:

Topic: To perform & practice the Mallory’s Phosphotungstic Acid Haematoxylin (PTAH)

Study Materials:

https://www.labce.com/spg531616_phosphotungstic_acid_hematoxylin_ptah_staining___c.aspx

https://en.wikipedia.org/wiki/Phosphotungstic_acid-haematoxylin_stain

Session 22

Topic: Introduction of FNAC and its staining technique

Study Materials:

Video- https://www.youtube.com/watch?v=JTYJBNxeTH8

Slide share- https://www.slideshare.net/DRKALPAJYOTI/fine-needle-aspiration-cytology-fnac

Session 23

Topic: (PAP stain and other special stain) for Paraffin Infiltrated and Embedded tissue

Study Materials: 

Protocol- https://www.ihcworld.com/_protocols/special_stains/papanicolaou_stain.htm

Video-https://www.youtube.com/watch?v=lmAGrOE49C0&t=27s

Session 26

Topic: Demonstration of instruments used for dissection

Study Materials:

Slide share- https://www.youtube.com/watch?v=dubPIssRh8M

https://www.youtube.com/watch?v=4AophuoO8Ec

Session 27

Practice session:

Topic:  Demonstration of instruments used for disection

Study Materials:

Video: https://www.youtube.com/watch?v=dubPIssRh8M

Session 28

Topic: Use of Disinfectants & antiseptics

Study Materials:

Video- https://www.youtube.com/watch?v=wzQ8NJg98lo

Session 29

Topic: laboratory Reception and labeling of histological specimens

Study Materials:

Video- https://www.youtube.com/watch?v=Snvi6tmQ-2M

http://www.pathology.uci.edu/services/specimen-labeling-requirements.asp

Session 30

Topic: Preparation of various fixatives -Helly’s fluid

Study Materials:

https://theolb.readthedocs.io/en/latest/histology/hellys-fluid-fixation.html

Video- https://www.youtube.com/watch?v=_ltg2cHsSWo

Session 31

Topic: Preparation of various fixatives -  Zenker’s fluid

Study Materials:

https://theolb.readthedocs.io/en/latest/histology/zenkers-fluid-fixation.html

Video- https://www.youtube.com/watch?v=_ltg2cHsSWo

 

 

Session 32

Topic: Preparation of various fixatives - Bouin’s fluid

Study Materials:

https://theolb.readthedocs.io/en/latest/histology/bouins-fluid-fixation.html

Video- https://www.youtube.com/watch?v=_ltg2cHsSWo

Session 33

Topic: Preparation of various fixatives - Corney’s fluid

Study Materials:

https://theolb.readthedocs.io/en/latest/histology/carnoys-fluid-fixation.html

Session 34

Topic: Preparation of various fixatives -10% Neutral formalin

Study Materials:

https://www.sigmaaldrich.com/catalog/product/sigma/ht501128?lang=en&region=IN

Session 35

Topic: Preparation of various fixatives -Formal saline

Study Materials:

http://diyhpl.us/~bryan/irc/protocol-online/protocol-cache/formsal.html

Session 36

Practice Session:

Topic: To perform embedding and casting of block

Study Materials:

Video- https://www.youtube.com/watch?v=Qsr9k9JY3FE&t=6s

https://www.youtube.com/watch?v=xIyXA3c3oxU

Session 37

Practice Session:

Topic: To process a bone for decalcification

Study Materials:

https://www.leicabiosystems.com/knowledge-pathway/an-introduction-to-decalcification/

Session 38

Topic: To prepare 70% alcohol from absolute alcohol

Study Materials:

Video- https://www.youtube.com/watch?v=TvQFDXQ9qKc&t=52s

https://www.youtube.com/watch?v=TvQFDXQ9qKc

Session 39

Practice Session:

Topic: Processing of tissue by manual and automated processor method

Study Materials:

Video- https://www.youtube.com/watch?v=kG5i6FVevc8

Session 40

Practice Session:

Topic: To demonstrate various part and types of microtome

Study Materials:

Video- https://www.youtube.com/watch?v=Oo_O1fZkQqc

https://www.youtube.com/watch?v=Oo_O1fZkQqc

Session 41

Practice Session:

Topic: learn sharpening of microtome knife (Honing and stropping technique) To perform section cutting

Study Materials:

Video- https://www.youtube.com/watch?v=Ad7DWKDUuhQ

Session 42

Practice Session:

Topic: learn mounting of stained smears

Study Materials:

https://www.nationaldiagnostics.com/histology/article/mounting-tissue-sections

Session 43

Practice Session:

Topic: To practice attachment of tissue sections to glass slides

Study Materials:

Video- https://www.youtube.com/watch?v=o2hajvoPF04

https://www.youtube.com/watch?v=Qsr9k9JY3FE

Session 44

Practice Session:

Topic: To learn using tissue floatation bath drying of sections in incubator (37 degree C)

Study Materials:

Video- https://www.youtube.com/watch?v=Qsr9k9JY3FE

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