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Tissue Fixation Techniques Guide

Formalin is the most widely used fixative for histological specimens. It preserves tissue by stopping all cellular activity and hardening the tissue. The optimal pH range for fixatives is neutral to stabilize cellular activity without distortion. The duration of fixation varies depending on the thickness and type of tissue, with thicker specimens requiring longer fixation. Proper fixation is a crucial step for histotechnique to allow proper orientation and staining of tissues.

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Chie Hisugan
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50 views5 pages

Tissue Fixation Techniques Guide

Formalin is the most widely used fixative for histological specimens. It preserves tissue by stopping all cellular activity and hardening the tissue. The optimal pH range for fixatives is neutral to stabilize cellular activity without distortion. The duration of fixation varies depending on the thickness and type of tissue, with thicker specimens requiring longer fixation. Proper fixation is a crucial step for histotechnique to allow proper orientation and staining of tissues.

Uploaded by

Chie Hisugan
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as DOCX, PDF, TXT or read online on Scribd

FIXATION

- Slightly acidic & slightly alkaline (buffered)


- Autotech – automated until infiltration
Fixative – chemicals; crucial step for
- Muscles – tissue basophils
histotehnique
Aim:
- The thicker the specimen, the longer the
procedure will take
- Float – impartial fixation

-CHEMICAL CONSTITUENT: carbohydrates,


proteins, lipids
-Soft: deform
- To allow the proper orientation of tissues - Hypertonic solution – cause shrinkage
during embedding - Hypotonic solution – cause swelling
- Preserving the tissue – stopping all chemical
cellular activities - Buffered formalin 2-6 hrs
- Harden tissue by stabilizing - Inhibit cells –
- Additive – whole tissue
- Non-additive – alterate – body fluids, aspirate
- Alcoholic fixative – dehydrating and fixative
agent

- Prevent immediate autolysis


- Duration varies; decrease duration = apply heat
- Compatible mostly to impregnation,
dehydration, etc.
- Very flexible
- Formalin (10%) widely used – stock solution
or concentrated – recommended to buffer to pH7
– cheap; compatible with many stains; almost no
interference
- Precipitate – reacts

- Friable tissues: bone marrow, other soft tissues


- Resistant to damage, disrupt the swelling
- Antibacterial
- 4&5, Fixative – increase the capacity or
infinity to absorb stains

- Nuclear fixatives – high affinity to acids


- Acetone & ethanol – cannot fxn well without
water - Optimal pH range neutral to stabilize cellular
activity
- Stable for the processing
- Longer because of buffer process (Ammonium
oxalates)
- Minimum with no distortion at all

- formol – mas mabilis magpenetrate


- mercury – open pores for increased penetration
of fixatives
- for lipids
- silver reticulin – nerve tissues
- Alcoholic formalin
- Nitric acid – very unstable element
- microincineration techniques
- Mucoidal – mahirap
- Uneven hardening of tissues

- Zenker – spleen or kidney


- Heidenhain – no need for washing; excellent
fixative for sliver impregnation techniques;
considerable tissue wrinkages
- B5 – 1 to 2 hrs; soft tissues, body fluids;
mabilis tumigas

- for electron microscopy – recommended:


frozen section
- recommended: very small specimen
- half hour to 4 hours fixation
- stable; pleasant fumes ( not irritating)
- expensive; volatile
- specimen w/ this must be kept in ref

- thin and ultra thin


- cryostat procedures - formalin with aldehyde group = reactive –
should use postchroming
- Orth’s – necrotic

- remedy =
- easy penetrate
- faster hardeninh
- highest affinity for acidic dyes
- nuclear stains
- De-zenkerization – black deposits

- bouins – brilliant tissue staining


- not for individual fixative fumes can cause blindness
- acidic osmium tetroxide – volatile than alcohol; bilis
mag evaporate; fumehood
1. no need for 20:1; there’s no acetic acid

- Overfix – overlyharden and shrinks

acetone : negri bodies - measles


pathognomonic
recommended for frozen pH – fixing – heat fix
bacterial smear – direct heat, indirect heat
(microwave, stove)

ethanol – can alter

new – immunochemistry tests


- washout - after mordant, after alcohol, after
counterstaining

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