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Calcification
Term applied to tissues which have been
infiltrated with calcium salts
Decalcification
Process of removing calcium ions or lime salts
from the organic cellular matrix, calcified
collagen and surrounding tissue of the bones.
Principle of Decalcification
Insoluble calcium salt are converted into
soluble calcium salts by the action of
decalcifying agent so that the tissue
become soft.
Result:
Decalcification is complete when no precipitate or
turbidity is observed
If slotuion turns turbid, the solution should be changed.
Chemical Test Procedure 2:
Step Action
1 Take 5 mL of decalcifying fluid
2 Add litmus paper/pH paper with magnetic stirrer
3 Add ammonium hydroxide, drop by drop, until indicators is
neutral (pH 7)
4 Add 5 ml ammonium oxalate. Shake well
5 Stand for 30 minutes
Result:
If white precipitate (calcium hydroxide) forms after adding NH4OH large
quantity of Ca++.
If clear, proceed with step 4. if precipitation occurs, Ca++ is present (small
amt)
If solution remains clear (step 5) decalcification is complete.
Treatment of Decalcified
Specimen
Chemical Neutralization
Immersed in lithium carbonate solution
Rinsing with water
30 mins small bones
1-4 hours large/dense bones
Treatment with alcohol (70%)
Decalcified with EDTA
Tissue Softeners
For unduly hard tissues that may damage
the microtome knife
Perenyis fluid tissue softener and
decalcifying agent
4% aqueous phenol
Molliflex
2% HCl
1% HCl in 70% alcohol