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What Are Characteristics of An Ideal Fixative?

An ideal fixative should maintain clear morphological features and consistent staining over long periods of storage. It should preserve tissue composition and structure while inducing minimal shrinkage or swelling. An ideal fixative also prevents autolysis and bacterial growth, permits molecular analysis, and is useful for various tissue types. It should fix and penetrate tissues rapidly while being compatible with automated processors, disposable, and cost effective. Physical fixation methods include heat, microwave, freeze-drying, and freeze substitution. Heat accelerates other fixation but microwaving tissue in formalin produces dangerous vapors. Freeze-drying removes water to study soluble materials while freeze substitution removes water through dissolution without denaturing proteins. These physical methods are primarily used for research

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1K views2 pages

What Are Characteristics of An Ideal Fixative?

An ideal fixative should maintain clear morphological features and consistent staining over long periods of storage. It should preserve tissue composition and structure while inducing minimal shrinkage or swelling. An ideal fixative also prevents autolysis and bacterial growth, permits molecular analysis, and is useful for various tissue types. It should fix and penetrate tissues rapidly while being compatible with automated processors, disposable, and cost effective. Physical fixation methods include heat, microwave, freeze-drying, and freeze substitution. Heat accelerates other fixation but microwaving tissue in formalin produces dangerous vapors. Freeze-drying removes water to study soluble materials while freeze substitution removes water through dissolution without denaturing proteins. These physical methods are primarily used for research

Uploaded by

Samra
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© © All Rights Reserved
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What are characteristics of an ideal fixative?

1. The major objective of fixation in pathology is to maintain ​clear and consistent


morphological features​. It should support high quality and consistent staining with
H&E, both initially and after storage of the paraffin blocks for at least a decade.
2. The ​local chemical composition of the tissue​ must also be maintained
3. It should induce ​minimal shrinkage or swelling.
4. It should ​prevent autolysis and bacterial putrefaction.
5. Fixatives should also permit the ​recovery of macromolecules including proteins,
mRNA, and DNA from fixed and paraffin-embedded tissues without extensive
biochemical modifications.
6. It should be ​useful for a wide variety of tissue types, including fat, lymphoid and
neural tissues.
7. It should preserve ​small and large specimens
8. It should support histochemical, immunohistochemical, in situ hybridization and other
specialized procedures.
9. The fixative should ​penetrate and fix tissues rapidly
10. It should have a ​shelf life of at least one year.
11. It should be ​compatible with modern automated tissue processors​.
12. It should be ​readily disposable or recyclable.
13. It should be ​cost effective.

What are different physical methods of fixation? Also mention


their advantages and disadvantages?
Heat fixation​:
Mechanism: Boiling or poaching an egg precipitates the protein. Each component is less soluble
in water after heat fixation than the same component of a fresh egg.

Advantage: Heat is primarily used to accelerate other forms of fixation. Picking up a frozen
section on a warm microscope slide, both attaches the section to the slide and partially fixes it
by heat and dehydration

Microwave fixation:
Mechanism: Same as heating
Advantage: Microwave heating can reduce times for fixation of some gross specimens and
histological sections from more than 12 hours to less than 20 minutes.
Commercial glyoxal based fixatives which do not form vapors when heated at 55°C have been
introduced as an efficient method of microwave fixation.
Disadvantage: Microwaving tissue in formalin results in the production of large amounts of
dangerous, potentially explosive vapors.

Freeze-drying

Advantage: Freeze-drying is a useful technique for studying soluble materials and small
molecules.

Mechanism: Tissues are cut into thin blocks, immersed in liquid nitrogen and the water removed
in a vacuum chamber at −40°C. The tissue can be post-fixed with formaldehyde vapor.

Freeze substitution:
Mechanism: When specimens are immersed in acetone or alcohol at −40°C, this slowly
removes water through dissolution of ice crystals and the proteins are not denatured. Bringing
the temperature gradually up to 4°C will complete the fixation process.

Advantage: These methods of fixation are used primarily in the research environment.

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