HISTOPATH DECALCIFATION

Question 1

Removal of Calcium or lime salts from bones or calcified tissues following fixation

Answer 1

DECALCIFICATION

Question 2

_____ solutions decalcify more rapidly but may destroy ____

Answer 2

more concentrated acid solutions ; tissue

Question 3

decalcification ratio

Answer 3

20:1

Question 4

____ hastens decalcification

Answer 4

heat and agitation

Question 5

decalcification duration

Answer 5

1-2 days

Question 6

tissue in decalcification

Answer 6

bones and teeth

Question 7

criteria for decalcifying agent

Answer 7

Complete removal of ca salts
No damage to tissue
Lack of harmful effect on staining reactions
Speed for removal of Ca salts

Question 8

types of decalcifying agents

Answer 8

Acid
Chelating agent
Ion exchange resins
Electrophoresis

Question 9

Widely used for routine purpose

Answer 9

Acid

Question 10

most common and a rapid decalcifying acid agent

Answer 10

5-10% Nitric Acid

Question 11

5-10% Nitric Acid time

Answer 11

12 to 24 hours

Question 12

Acid can be removed by

Answer 12

70% alcohol or formaldehyde

Question 13

Imparts a yellow color due to formation of Nitrous Acid

Answer 13

nitric acid

Question 14

5-10% nitric acid is used for

Answer 14

For urgent biopsies, for needle and small biopsies
For large and heavily mineralized cortical bone specimen

Question 15

Yellow color may impair staining reaction
Produce less tissue distortion than 10% aqueous nitric acid

Answer 15

FORMOL-NITRIC ACID

Question 16

FORMOL-NITRIC ACID is used for

Answer 16

For urgent biopsies
For nuclear staining

Question 17

FORMOL-NITRIC ACID time

Answer 17

Time: 1-3 days

Question 18

Decalcifies and soften tissues at the same time
Nuclear and cytoplasmic staining is good
Slow decalcifying agent for dense bones

Answer 18

PERENYI’S FLUID

Question 19

Most rapid decalcifying agent
Nuclear staining is poor

Answer 19

PHLOROGLUCIN-NITRIC ACID

Question 20

PHLOROGLUCIN-NITRIC ACID time

Answer 20

12-24 hours

Question 21

Yellow color must be neutralized with

Answer 21

5% Sodium sulfate

Question 22

in PHLOROGLUCIN-NITRIC ACID, When decalcification is complete..

Answer 22

the acid must be removed by 3 changes of 70% to 90% ethanol, when sections are cut the slides are brought to water and placed in 1% aqueous lithium carbonate for 1 hour and washed for 15 minutes then stained.

Question 23

PERMITS good cytologic staining
Moderately rapid decalcifying agent
Does not require washing out

Answer 23

HYDROCHLORIC ACID

Question 24

HYDROCHLORIC ACID is used for

Answer 24

For teeth and small pieces of bone

Question 25

The extent of decalcification cant be measured by a

Answer 25

chemical test

Question 26

Fixative and decalcifying agent Excellent nuclear and cytoplasmic staining

Answer 26

Formic Acid

Question 27

Formic Acid is used for

Answer 27

For small pieces of bones and teeth For immunohistochemical staining

Question 28

Formic acid Requires..

Answer 28

neutralization with 5% Na sulfate and washing out to remove the acid from the tissue

Question 29

Good nuclear staining Does not require washing Weak agent, not used for dense tissues and suitable for small spicules of bone Not recommended for urgent biopsies

Answer 29

TRICLOROACETIC ACID

Question 30

Weak decalcifying agent Suitable for minute bone spicules

Answer 30

SULFUROUS ACID

Question 31

Used ad fixative and decalcifying agent For minute bone spicules Nuclear staining with hematoxylin is inhibited Chromic aid is environmental toxin Highly corrosive to skin, mucous membrane carcinogenic

Answer 31

CHROMIC ACID (Flemming’s Fluid)

Question 32

COMBINE WITH Calcium salts and other salts to form complexes and to facilitate removal of calcium Excellent for immunohistochemical or enzyme staining and for electron microscopy

Answer 32

Chelating Agents

Question 33

common chelating agent

Answer 33

EDTA

Question 34

Chelating Agents duration

Answer 34

1-3 weeks for small specimens 6-8 weeks or longer for dense bones

Question 35

in Chelating Agents, the pH is adjusted to.

Answer 35

7 to 7.4

Question 36

Ammonium form of polysterene resin hastens decalcification by removing calcium ions from formic acid-containing decalcifying solutions Not recommended for fluids containing mineral acids such as Nitric acid or Hydrochloric acid Cellular detail is well-preserved Extent of decalcification can be measured by routine chemical test

Answer 36

ION EXCHANGE RESIN

Question 37

Is a process whereby positively charged calcium ions are attracted to negative electrode and subsequently removed from the decalcifying solution.

Answer 37

ELECTROPHORESIS

Question 38

in ELECTROPHORESIS, the time required for decalcification is thereby..

Answer 38

shortened due to the heat and electrolytic reaction produced in the process.

Question 39

Acid can be removed from tissues or neutralized chemically by immersing the decalcified bone in either saturated lithium carbonate solution or 5-10% aqueous Na bicarbonate solution for several hours

Answer 39

POST-DECALCIFICATION

Question 40

in post-decalcification ___ is used for rinsing

Answer 40

running tap water

Question 41

how long is running tap water

Answer 41

30 min for small; 1-4 hrs for larger specimen

Question 42

Measuring Extent of Decalcification

Answer 42

Physical or Mechanical Test X-Ray or Radiological Method Chemical Method/Calcium Oxalate Test

Question 43

Done by touching or bending the tissue Pricking the tissue with fine needle or probe

Answer 43

Physical or Mechanical Test

Question 44

Ideal, most sensitive and expensive Detects smallest focus of Calcium

Answer 44

X-ray or Radiological Method

Question 45

Detects Calcium in acid solution by precipitation of insoluble Calcium hydroxide or Calcium oxalate Cloudiness/ppts indicates Ca is present

Answer 45

Chemical Method/ Calcium Oxalate Test