FIXATION Flashcards
Post martem devomposition or autolysis occurs due to the action of what enzyme
Hydrolytic enzymes
Fixation prevents autolysis by inactivating by what enzyme
Lysosomal enzyme
2 types of mechanism
Additive fixation
Non additive fixation
Mechanism whereby the chemical constituent of the the fixative is taken in
Forming crosslinks or molecular complexes
Additive fixation
Mechanism whereby altering the tissue composition
Upon removal of water to to hbonds, new cross links are established
Acetone and alcohon fixatives are examples
Non-Additive fixation
Formalin heated at 60C
Rapid fixation for very urgent biopsy specimens
Formalin heated at 100C
For tuberculosis
Volume of fixative used
10-25 times the volume of the tissus
Maximum effectiveness
- 20 times the volume of specimen
For museum
- not less than 50-100 times the specime
Penetration of fixatives in the tissue at the rate of
1mm/hour
Intensifies hematoxylin staining?
Inhibits?
Formaldehyde
Osmium tetroxide
Fixative that react with viruses
Mercuric chloride
Commonly used fixatives for nucleic acids
MEC
methanol
Ethanol
Carnoys
Involves thermal coagulation
For frozen tissue sections (urgent biopsies)
Preparation of bacteriologic smear
Heat fixation
Process of placing an already fixed tissue in a second fixative
Acting as a mordant
Secondary fixation
Stain MASSON’S TRICHROME
for connective tissue
MALLORY’s ANILINE BLUE STAIN
for collagen
PHOSPHOTUNGSTIC ACID HEMATOXYLIN
for striated muscles
Secondary fixation whereby the primary fixation is placed in Potassium dichromate
Post-chromatization
Removing of excess fixative in order to improve staining and remove artifacts
WASHING-OUT
Tap water used to remove
COF
chromates
Osmic acid
Formalin
Used to remove excess amount of picric acids
50-70% alcohol
Used to remove excess mercuric chloride
Alcoholic iodine
Fixation is retarded by
Size and thickness Mucus Fat Blood Cold temp
Fixation enhanced by
Size and thickness
Agitation
Heat
Volume of fixatives if expensive osmium tetroxide is used
5-10 times the volumes only
Floating of air filled organs like lungs may be avoided
Covered with several layer of gauze g
Before hollow organs like stomach and intestines immersed, do:
Packed with cotton soaked fixatives
What to do to prevent brain from flattening
INTRAVASCULAR PERFUSION
Washing out of bloos with RINGERS LACTATE
Hard tissues
-cervix, uterine, fibroids, hyperkeratotic, nails
What u should do? For tissue softener?
LENDRUM’S METHOD
Washed out with running water overnight
Add 4% acqueous phenol solution for 1-3 days
Factors considered when choosing the right fixatives
Need for immediate exami Tissue structure Type of tissue Type of section Staining technique
For glycogen demontration
Picric acid
Glycogen preservation
Glycogen polarization
Alcohol fixative
Strong oxidizing agent
Chromic acid
Osmium tetroxide