HISTOTECHNOLOGY – II & Cytology BMLS 605-18

HISTOTECHNOLOGY – II & Cytology

 BMLS 605-18

Theory

Aims and Objectives:

The students will learn about various staining procedures for demonstration of different substances & various cytological investigations. The students will learn about special staining procedures & handling & testing of various cytological specimens.


HISTOPATHOLOGY

https://rajswasthya.nic.in/RHSDP%20Training%20Modules/Lab.%20Tech/Histo/Introduction.pdf

STAINING

https://rajswasthya.nic.in/RHSDP%20Training%20Modules/Lab.%20Tech/Histo/Chapter%206.pdf

HISTOCHEMISTRY

https://www.du.edu.eg/upFilesCenter/exStore/sci/1584370154.pdf



1. Cryostat sectioning, its applications in diagnostic histopathology.

https://drive.google.com/file/d/1BPHfo5n_LoDnUWCOW0ZzGEF45jo9fWps/view?usp=sharing

https://www.slideshare.net/Mahmoud19905/frozen-section-and-cryostat

https://www.pathologyoutlines.com/topic/methodsfrozen.html


2. Special Staining Procedures for detection of

a. Connective tissue elements, Trichrome staining, muscle fibers, elastic, reticulin fibres, collagen fibres etc.        =       Special Stains

b. Metachromatic staining such as toludine blue on frozen sections

c. Principles of metal impregnation techniques.

d. Demonstration and identification of minerals and pigments, removal of pigments/artifacts in tissue sections.



1) Masson Trichrome stain: https://www.slideshare.net/zahoor061/massons-trichrome-stain
2) weigert- van gieson stain:

     https://www.slideshare.net/aghara33/special-stain-in-histopathology     ( slide no. 41 to 125)


Verhoeff’s solution needs to be fresh each use due to rapid oxidation. Do not over differentiate in the ferric chloride as the dye molecules love this stuff equally as well as they do the tissue molecules they are bound to and will disconnect from the tissue easily. If this happens you will have to start all over. Incubate for a few seconds then rinse quickly and view microscopically looking for the elastic tissue fibers. When they are black and easily differentiated from collagen and other tissue types, stop the process.

Likewise, do not incubate too long in the van Gieson as the picric acid in solution will continue to differentiate out the bound hematoxylin.


3) weigert’s Resorcin Fuchsin: https://webpath.med.utah.edu/HISTHTML/MANUALS/WEIGERTS.PDF

SPECIAL STAINS

https://www.leicabiosystems.com/en-in/knowledge-pathway/special-stains-which-one-why-and-how/


3. Demonstration of Proteins & nucleic acids.


4. Demonstration of Carbohydrates, lipids, fat & fat like substances.


5. Demonstration of bacteria and fungi in tissue section.


6. Tissue requiring special treatment i.e. eye ball, bone marrow, muscle biopsy, under-calcified or uncalcified bones, whole brain, whole lungs including other large organs.


UNIT 6:-  DEACLCIFICATION OF BONE, EYE BALL
https://nios.ac.in/media/documents/dmlt/HC/Lesson-17.pdf


STAINS FOR EYE 
https://www.ncbi.nlm.nih.gov/pmc/articles/PMC3846726/


7. Enzyme histochemistry : Diagnostic applications & demonstration of Phosphatases, Dehydrogenases, Oxidases & Peroxidases.

https://drive.google.com/file/d/1p0HIIPXWDeNBqzlY2gluUIMr7yZXm6IH/view?usp=sharing

PRP = Primary Reaction Product, the product of the reaction of an enzyme on a substrate.

FRP = Final Reaction Product, the product of an insoluble uncolored P.R.P. which has been rendered colored or opaque.


https://docs.google.com/presentation/d/e/2PACX-1vRoLNMCfH4WoIZjT6hdCUJIx762pzTu5XuG7ZoMGYANVHJVLBBo5TiJtqU_mKsrBQ/pub?start=false&loop=false&delayms=3000


8. Vital staining.

https://www.slideshare.net/AnilYadav6/vital-staining-ppt-notes

https://www.peertechzpublications.org/articles/ACP-5-117.pdf


9. Neuropathological techniques.

https://core.ac.uk/download/pdf/268450394.pdf

STAINING:
Nissl Staining         http://www.ihcworld.com/_protocols/special_stains/nissl.htm
Luxol Fast Blue Stain    http://www.ihcworld.com/_protocols/special_stains/fast_blue.htm
Kluver-Barrera Method    https://www.emsdiasum.com/microscopy/technical/datasheet/26681.aspx
BODIAN'S METHOD       https://webpath.med.utah.edu/HISTHTML/MANUALS/BODIAN.PDF  
GLIAL FIBERS       https://webpath.med.utah.edu/HISTHTML/MANUALS/HOLZER.PDF
Gallyas Silver Stain    https://www.protocolsonline.com/histology/dyes-and-stains/neurohistology/gallyas-silver-stain/


10. Museum techniques.

https://nios.ac.in/media/documents/dmlt/HC/Lesson-20.pdf

https://www.slideshare.net/shashishara/museum-techniques-75256254

https://www.slideshare.net/shashishara/museum-techniques-75256254#:~:text=%EF%82%97%20Fixation%20arrests%20autolysis%20and,Kaiserling%20technique%20and%20his%20modifications.


11. Aspiration cytology principles, indications & utility of the technique with special emphasis on role of cytotechnologist in FNAC clinics, fluid cytology.

1) https://www.exeterlaboratory.com/cellular-pathology/fine-needle-aspiration-fna-cytology/#:~:text=Principle%20of%20Examination,does%20not%20preserve%20histological%20architecture.

2)FNAC:- https://www.slideshare.net/DRKALPAJYOTI/fine-needle-aspiration-cytology-fnac

3) BODY FLUID:- https://www.slideshare.net/peddanasunilkumar/body-fluids-119314504


12. Exfoliative cytology (Papanicolaou technique for the staining of cervical smears.)

1. https://www.slideshare.net/ShaminJoshi/exfoliative-cytology-64853836

2.  https://www.slideshare.net/ImranaTanvir/cytology-exfoliative


13. Introduction and importance of Autoimmune disorders, HLA typing, T & B Cells, ANA, ANCA and TMA

1) https://www.britannica.com/science/autoimmune-disease
2) https://www.slideshare.net/drnisha22/hla-92546736
3) https://www.slideshare.net/debbarma1989/hla-typing-and-its-role-in-tissue-transplantation
4) https://www.slideshare.net/RehnumaKawsar/tcell-45647452
5) https://www.slideshare.net/CarolineKarunya/b-cellimmunology
6) https://medlineplus.gov/lab-tests/ana-antinuclear-antibody-test/
7) https://medlineplus.gov/lab-tests/antineutrophil-cytoplasmic-antibodies-anca-test/

TMA:- https://unckidneycenter.org/kidneyhealthlibrary/glomerular-disease/thrombotic-microangiopathy-tma/



14. Cancer immunology.

1) https://www.slideshare.net/MMASSY/tumour-immunology-leture-notes
2) https://www.slideshare.net/hvphitesh/cancer-immunology-41021988


15. Tissue typing for kidney transplant.

1)https://www.gosh.nhs.uk/conditions-and-treatments/procedures-and-treatments/tissue-typing-kidney-donation/#:~:text=Contrary%20to%20what%20it%20sounds,of%20viruses%20from%20the%20body.

2)https://onlinelibrary.wiley.com/doi/pdf/10.1111/nep.13637




EXTRA


Immunohistochemistry Protocol for Paraffin embedded Tissue Sections VEDIO LINK:-

https://www.youtube.com/watch?v=XMjGZHEG4cY


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