LESSON 3: DECALCIFICATION (2)

Question 1

excellent bone decalcifying agent for immunohistochemistry and enzyme studies
✓ acts as both decalcifying agent and water softener
✓ does not interfere with staining
✓ does not distort tissues and enzymes
✓ available in 2 formulations

Answer 1

EDTA

Question 2

WHAT IS THE OPTIMUM PH BINDING OF EDTA?

Answer 2

PH 8

Question 3

What pH inhibits calcium binding

Answer 3

pH 3

Question 4

What pH allows binding and does not destroy tissue components

Answer 4

pH 7.0 to 7.4

Question 5

✓ uses ammonium-sulfonated polysterene + formic acid
✓ volume of the acid solution: 20-30 times the volume of the sample
✓ Resin and Formic Acid (RAF)

Answer 5

ION EXCHANGE RESIN

Question 6

✓ positively charged calcium ions are attracted to a negative electrode
✓ uses heat and electrolytic reaction
✓ dependent on electricity for the removal of calcium
✓ temperature: 30 to 45 degrees Celsius
✓ uses 90%/88% formic acid and concentrated HCl as the acid solution

Answer 6

ELECTROPHORESIS

Question 7

✓ done by touching or bending to determine consistency or pricking with fine NEEDLE OR PROBE
✓ vague and not a reliable method

Answer 7

PHYSICAL TEST

Question 8

✓ calcium and mineral salts produce opaque areas
✓ very expensive, most ideal method, most sensitive and most reliable method
✓ DO NOT use for mercuric chloride fixed tissues
✓ EQUIPMENT: Faxitron and Kodak X-OMAT X-ray film

Answer 8

RADIOLOGIC TEST

Question 9

Simple, reliable and convenient method for routine purposes

Answer 9

CHEMICAL TEST

Question 10

(+) precipitate = (+) for calcium = ?

Answer 10

INCOMPLETE DECALCIFICATION

Question 11

✓ surface blocks submerged for 1 to 2 hours
✓ tissues immersed for 12 to 24 hours

Answer 11

Perenyi’s Fluid

Question 12

may cause swelling or make tissues soapy

Answer 12

Molliflex