metodo de grimelius

8/7/2019 Metodo de Grimelius

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SURGICAL PATHOLOGY - HISTOLOGY Dat e:

STAINING MANUAL - SPECIAL CELLS AND TISSUES Page: 1 of 3

GRI MELI US - PA SCUA L' S MODI FI ED - A RGYROPHI L CELL S

PURPOSE: These cells are found in larger numbers in the same general

locat ions as the argent aff in cells. Argent aff in cells will have a posit ivereact ion with the argyrophil t echniques, but t he argyrophil cells will not

react with the argentaffin techniques. This procedure is used for the

dif f erent iat ion of carcinoid t umors.

PRI NCI PLE: Argyrophil cells are capable of being impregnated wit h

silver, but need a reducing agent t o reduce t he silver t o a visible metallic

silver. While argent aff in cells have bot h t he abilit y of being impregnated

wit h silver and reducing t he silver.

CONTROL: Tissue known t o cont ain argyrophil granules or small int est ine.

FIXATION: 10% formalin

TECHNI QUE: Cut paraf f in sect ions 4 .

EQUI PMENT: Acid c leaned glassware, pH met er, microw ave oven.

REAGENTS:A c i du l at ed Wat er :

Dist i l led w at er 1000 .0 ml

0.3 % Cit ric acid 50 .0 ml

Add the 0.3% citric acid, drop by

drop, unt il a pH has been

established at 4.2. Make fresh,

discard aft er use.

CAUTI ON: Av oi d c ont ac t and i nha l at i on.

1 % Si l v er Ni t r at e:

Silver nit rat e 0.5 gm

Acidulat ed wat er 50 .0 ml

Make fresh, discard af t er use.


Reduc i ng Sol ut i on :

Sodium sulfit e, anahydro us 2.5 gm

Hydroquinone 0.5 gm





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SPECIAL CELLS AND TISSUES

GRI MELI US Page: 2 of 3

SAFETY: Wear gloves, goggles and lab coat . Avoid cont act and inhalat ion.

Silver nit rate: severe skin and eye irrit ant . Oxidizer. Ingest ion will

produce violent gast rointest inal discomfort . Possible carcinogen:equivocal t umorigenic agent .

Sodium sulf it e; toxic on ingest ion. can irrit ate t he st omach. Irrit ant t o

skin, eyes and respirat ory syst em.

Hydroquinone; irritant t o skin and eyes. Toxic by inhalat ion and ingest ion.

Chronic expo sure has caused bladder cancer in m ice.

PROCEDURE:

1. Deparaff inize and hydrat e to acidulat ed water.

2. * 1% Silver solut ion, microwave 80 power, 5 0 seconds. Save solut ion.

3. Rinse slides in hot acidulat ed wat er.

4. *Heat Reducing solut ion, 55 seconds, Hi power. Place slides in hot

solut ion for 3 -5 minut es. Save solut ion.

5. Rinse in 3 changes of hot acidulat ed wat er.

6. * 1% Silver solut ion, microwave 80 power, 30 seconds.

7. Rinse in hot acidulated wat er.

8. *Reheat Reducing solut ion, 15 seconds, Hi power. Place slides in hot

solut ion f or 1- 2 m inut es. Check microscopically f or black cyt oplasmic

granules, repeat st eps 6 t hrough 8 if needed.9. Rinse in t ap wat er f or several minut es.

10 . Dehydrat e, clear, and coverslip.

*Convent ional Met hod:

2. Silver solut ion, 60 C , 1 hour.

4. Heated Reducer, 58C, 5 minutes.

6. Silver solut ion, 5 8 C, 1 0 minut es.

8. Heat Reducer, 58 C, 1 0 minutes.

RESULTS:Arg y ro ph il li c c el ls b lac k

Background yellow t o gold


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SPECIAL CELLS AND TISSUES

GRI MELI US Page: 3 of 3

REFERENCES:

Pascual JS, A New Met hod for Easy Demonst rat ion of Argyrophil cells.

St ain Tech, 51 :231 -23 5, 19 76

Sheehan D, Hrapchak B, Theory and pract ice of Histot echnology, 2nd Ed,

19 80 , pp 27 8-2 79 , Batt elle Press, Ohio

Crookham,J, Dapson,R, Hazardous Chemicals in t he Hist opat hology

Laboratory, 2nd ED, 19 91, Anat ech

Prepared: By:

Approved: By:

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PROCEDURE CARD

GRI MELI US - PA SCUA L' S MODI FI ED - A RGYROPHI L CELL S

CONTROL: Tissue known t o cont ain argyrophil granules or small int estine.

TECHNI QUE: Cut paraff in sect ions 4.

PROCEDURE:1. Deparaffinize and hydrat e t o acidulat ed water.

2. *1 % Silver solut ion, microwave 80 power, 50 seconds. Save solut ion.

3. Rinse slides in hot acidulated water.

4. *Heat Reducing solut ion, 55 seconds, Hi power. Place slides in hot solut ion for 3 -5

minut es. Save solution.

5. Rinse in 3 changes of hot acidulat ed water.

6. *1 % Silver solution, microwave 80 power, 30 seconds.

7. Rinse in hot acidulat ed wat er.

8. *Reheat Reducing solut ion, 15 seconds, Hi power. Place slides in hot solut ion for 1 -2

minutes. Check microscopically f or black cyt oplasmic granules, repeat steps 6 t hrough 8

if needed.

9. Rinse in t ap wat er for several minutes.

10 . Dehydrat e, clear, and coverslip.

*Convent ional Met hod:

2. Si lver solut ion, 60 C , 1 hour.

4. Heat ed Reducer, 58 C, 5 minutes.

6. Silver solut ion, 58 C, 10 minutes.

8. Heat Reducer, 58 C, 10 minutes.

RESULTS:Argyrophillic cells black

Background yellow t o gold

A c i dul at ed Wat er :

Dist illed wat er 1 00 0 .0 ml

0 .3 % Cit r ic acid 5 0 .0 ml

Add the 0.3% citric acid, drop bydrop, until a

pH has been est ablished at 4.2 . Make fresh,

discard after use.


1 % S i l v e r Ni t r a t e:

Silver nit rat e 0.5 gm

Acidulat ed wat er 5 0 .0 ml



Reduc i ng Sol ut i on:

Sodium sulfit e,

anahydrous 2.5 gm

Hydroquinone 0.5 gm




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