DECALCIFICATION Flashcards

1
Q

Injurious to the organic ground substance of the bone or other tissues

A

Acid solutions

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2
Q

How often are acid solutions changed

A

-once or twice until decalcifiyis completed

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3
Q

How often are acid solutions changed

A

-once or twice until decalcifiyis completed

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4
Q

Reagent volume and tissue ratio for decalcification

A

20:1

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5
Q

Ideal decalcification time

A

24-48 hrs

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6
Q

Decalcification time for dense bones

A

14 days or longer

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7
Q

Optimum temp for decalcification

A

18-30C

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8
Q

Examples of agitation

A
  • low speed rotation
  • stirring
  • introduction of bubbles
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9
Q

Temp in which tissues will undergo complete digestion

A

55C

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10
Q

temp in which it impairs nuclear staining with van gieson’s

A

37C

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11
Q

temp in which it reduced effectiveness of trichrome and PAS

A

37C

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12
Q

percentage of preparing nitric acid

A

5-10%

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13
Q

most common and fastest decalcifying agent

A

nitric acid

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14
Q

components of nitric acid

A

formaldehyde or alcohol and chromic acid

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15
Q

causes yellow spontaneous yellow discoloration (HNO2)

A

nitric acid

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16
Q

remedy of yellow discoloration

A

-neutralize with 5% NaSO4 > wash for 12 hrs
- add 0.1% urea

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17
Q

decalcifying and water softener

A

EDTA

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18
Q

decalcifying and softener

A

perenyi’s

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19
Q

removal of calcium ions from a bone or calcified tissue through histological processes that makes them flexible and easier to cut

A

decalcification

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20
Q

3 main types of decalcifying agents

A
  • strong mineral acids
  • weaker organic acids
  • chelating agents
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21
Q

10% aqueous nitric acid solution formula

A
  • conc. HNO3
  • dH2O
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22
Q

decalcification time of aq HNO3 sltn 10%

A

12-24 hrs

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23
Q

formol HNO3 formula

A

-concd. HNO3
- strong formaldehyde 40%
- dH2O

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24
Q

decalcification time of formol-HNO3

A

1-3 days

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25
perenyi's fluid formula
-HNO3 - chromic acid - absolute ethyl alcohol
26
decal. time of perenyi's
2-7 days
27
phloroglucin-HNO3
- conc. HNO3 - Phloroglucin Nitric acid 10%
28
decal time of phloroglucin
12-24 grs
29
most rapid decalcifying agent so far, recommended for urgent cases
Phlorogucin _ HNO3
30
inferior compared to HNO3 in its role as a decal agent because of its slower action and greater distortion of tissue produced on the decalcified section
HCl
31
solution recommended for surface decalcification of tissue blocks
1% HCl, 70% alcohol
32
moderation of HCl recommended for teeth and small pieces of bones
von ebner's
32
fixative and decalcifying agent for post mortem research tissues
Formic acid
33
for small and large pieces of bones and gentler on tissues
formic acid
34
decalcification time for formic acid
2-7 days
35
recommended for autopsy materials, bone marrow, cartilage and tissues studied for research purposes
Formic Acid - sodium citrate
36
decalcification time for Formic acid - sodium citrate
3-14 days
37
T/F FA-Sc requires neutralization with 5% sodium sulfate
T
38
decal time for TCA
4-8 days
39
does not require washing out and is a weak decalcifying agent, not used for dense tissues, and is suitable for only small spicules of bones
TCA
40
very weak decal sltn suitable for minute pieces of bones
sulfurous acid
41
fixative and decal agent for minute pieces of bones
chromic acid/ flemming's
42
environmental toxin
flemming's
43
permits excellent nuclear and cytoplasmic staining and it does not produce cell or tissue distortion. However, action is too slow for routine purposes
Citric acid - citrate buffer
44
substances that combine with Ca ions and other salts to form weakly dissociated complexes and facilitate removal of Ca salt
Chelating agents
45
most common chelating agent
EDTA
46
commercial name of EDTA
versene
47
used as anticoagulant and H2O softener
EDTA
48
pH of EDTA
7-8
49
T/F Edta takes longer decalcification time - weeks rather than days
T
50
how much EDTA is needed for 1g of bones
2-4 oz
51
decal time of edta for small samples
1-3 weeks
52
decal time of edta for dense cortical bones
6-8 weeks
53
hastens decalcification by removing Ca ions by removing Ca ions from formic acid-containing decalcifying solutions, thereby increasing solubility from the tissue
Ion exchange resin
54
resin used for ion exchange resin
ammonium-sulfonated polystyrene
55
end point of ion exchange resin
- x-ray - physical method
56
reactivation of used resin
- N/10 HCl (2x) - dH2O (3x)
57
volume of ion exchange resin
20-30
58
process whereby positively charged Ca ions are attracted to a negative electrode and subsequently removed from the decalcifying solution
electrophoresis
59
sltn used for electrolytic decal
- 88-90% Formic acid - conc. Hcl - dH2O
60
time of decal is shortened d/t heat and electrolytic rxn produced in the process
electrophoresis
61
temp for electrophoresis
30-45C
62
post-decalcification treatment for washing out
running water: 30 mins to 1-4 hrs
63
post-decalcification treatment for neutralization
- 2% Li2CO3 - 5-10% Na Bicarbonate
64
post-decalcification treatment for frozen sectioning
* acid- decalcified tissues ---> water * store in formal - saline with 15% sucrose or * in phosphate-buffered saline with 15-20% sucrose
65
at what temp should frozen sectioning for post-decalcification done
4C
66
a method of dealing with small unexpected deposits of calcium that may be encountered in paraffin blocks
surface decalcification
67
may cause resistance of gratings sensation in surface decalcification
small foci
68
remedy for surface decal
- reome block - place facedown on a pad of cotton of gauze saturated with 10% HCl for approx 1 hr - rinse with HCl, chill, then section
69
other name for chemical test
Ca oxalate test
70
cannot be used when radiopaque metallic salts have been used
x-ray
71
x-ray cannot be used when _______ have been used
radiopaque metallic salts
72
reagents for chemical test
- 0.5 mL sat. aq. NH4 oxalate - 1% Na oxalate
73
ppt of chemical testing
- Ca oxalate - Ca hydroxide
74
tissue softener that causes tissue to swell and become soapy
molliflex
75
tissue softeners
- perenyi's - 4% aq. phenol - molliflex