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Fixation

Fixation ,aim of fixation, advantages, disadvantages

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safiyaashfaq72
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46 views

Fixation

Fixation ,aim of fixation, advantages, disadvantages

Uploaded by

safiyaashfaq72
Copyright
© © All Rights Reserved
We take content rights seriously. If you suspect this is your content, claim it here.
Available Formats
Download as PPTX, PDF, TXT or read online on Scribd
You are on page 1/ 19

PRESENTATION

PRESENTED BY: Dr.safia ashfaq


Introduction

 As soon as cells or tissues are removed from the body they


begin to die and undergo post- mortem changes.
 These changes may be autolytic or putrefactive.
* Autolysis: it Is a Self destructive process due to
the release of autolytic enzymes from the dead
cells.
 Putrefaction: Occurs due to the action of bacteria
that invade the tissue.
 It should prevent autolysis & putrefaction of the
cell.
 2. It should penetrate evenly and rapidly.
 3. It should harden the tissues
 4. Increase the optical differentiation of cells &
Aims of tissues
Fixation  5. Should not cause shrinkage or swelling of the
cells
 6. Must not react with the receptor sites & thus
must notinterefere with the staining procedure.
 7. It must be cheap and easily available.
Properties of an Ideal Fixative
 Prevents autolysis and bacterial decomposition.
 Preserves tissue in their natural state and fix all
components.
 Make the cellular components insoluble to reagent
used in tissue processing.
 Preserves tissue volume.
Avoid excessive hardness of tissue.​

Allows enhanced staining of tissue.​

Should be non-toxic and non-allergic


for user.​

Should not be very expensive.​


Mechanism / Action
of Fixative
 At the molecular level,
fixative have the
property of coagulating
proteins in the tissue,
through the formation
of crosslink's between
protein molecules
thereby keeping their
relation to each other.
Mechanism / Action of Fixative

 At the molecular level, fixative have the property of


coagulating proteins in the tissue, through the formation
of crosslink's between protein molecules thereby keeping
their relation to each other.
Classification o f Fixatives :

1) Physical Method of fixation

2)Chemical method of fixation.


 Heat fixation :
 The simplest form of fixation is heat.
Physical  Microwave Fixation:
method of  Microwave heating speeds fixation and can
fixation reduce times for fixation of some gross
specimens and histological sections from more
than 12 hours to less than 20 min.
 Freeze-Drying is a useful technique for
studying soluble materials and small
molecules; tissues are cut into thin sections,
Freeze-Drying immersed in liquid nitrogen, and the water is
and freeze removed in a vaccum chamber at -400c.
substitution :  The tissue can be post-fixed with
formaldehyde.
 Chemical fixation utilizes organic or non-
organic solutions to maintain adequate
morphological preservation.
 Chemical fixatives can be considered as
Chemical members of three major categories..
Fixation  1.coagulant
 2.Cross-linking
 3.Compound fixatives
 Formalin
 The most commonly used fixative is Formalin.
 It is prepared by mixing 40% Formaldehyde gas
in 100 w/v of distilled water.
Simple
 The resultant mixture is 100% Formalin.
Fixatives
 Routinely, 10% formalin is used which is
prepared by mixing 10 ml of 100% formalin in
90 ml of distilled wate
It fixes 4 mm thick tissue in 8 hours
OF ACTION
MECHANISM
It forms cross links between amino
acids of proteins thereby making
them insoluble.
 1.Rapid penetration
 2. Easy availability & Relatively cheap (Low
cost)
 3. Does not over harden the tissue
 4. Fixes lipids for frozen sections
ADVANTAGES
 5.It is relatively easy to prepare
:
 6.It allows subsequent use of most staining
procedures.
 7.frozen sections can be made with formalin
fixed tissue.
DISADVANTAGES:

 1.Irritant to the nose, the eyes and mucous membranes


 2. Formation of precipitate of paraformaldehyde which
can be prevented by adding 11-16% methanol.
 3. Formation of black formalin pigment, Acid
formaldehyde hematin.
 4.It causes shrinkage of collagen.
 5.it suspected to contain cancer producing agents
1-Temperature
2- Size of the specimen
3- Volume ratio
4- Duration of Fixation
5- Choice of fixatives
Factors
6-Penetration
affecting
7- Tissue Storage
fixation:-
8-Buffer & pH
9 - Osmolality
Glutaraldehyde

Fixatives used Osmium tetroxide


for Electron
Microscopy Formaldehyde & Glutaraldehyde mixture

Acrolein
Thank you

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