PH 122 LAB EXERCISE: TISSUE PROCESSING

PROCEDURE:
1. Cut a tissue sample with a dimension of 2cm x 2cm x 5mm. Hollow tissues should have cotton in
the lumen.
2. Wrap the tissues in gauze and tie with a thread.
3. Immerse the tissues in ascending concentrations of alcohol followed by two changes of xylene.
Follow the schedule below:
Dehydration
70% Alcohol
95 % Alcohol
95% Alcohol
Absolute Alcohol I

3h
1h
1h
1h
(Immerse in xylene to check for complete
dehydration. if xylene becomes turbid upon
immersion of tissues, place the tissue in second
absolute alcohol and transfer to xylene after an
hour. If xylene is clear, let the tissue stay for 1
hour)

Absolute Alcohol II
Clearing
Xylene I

1h
1h
(if xylene becomes turbid upon immersion of
tissues, return the tissues in absolute alcohol
and transfer to xylene after an hour)

Xylene II
1h
Infiltration
Paraffin I
1h
Paraffin II
1h
Embedding
4. Make a paper boat by cutting a 12cm x 9cm glossy paper and folding it to make a 9cm 3. Make sure
that the paper is hard enough to hold the solid paraffin wax.
5. Fill up to 1/3 of the boat with the molten paraffin wax. Allow a thin film of paraffin at the bottom
to partially solidify. This will only take a few seconds.
6. Lift the tissue from the final wax, with previously warmed forceps, and place in the bottom of the
mold. Orient the tissue properly on the mold by placing the side of the tissue from which it is desired
to take sections, faced down. Press down the tissue speciemen in the mold for a few seconds intuil it
is held by the cooling wax.
7. Pour additional molten paraffin in the mold until of the paper boat is filled or until the entire
tissue is covered with wax.
8. When the wax becomes partially solid, place the mold in a basin with ice or cold water to
immediately cool the wax and avoid crystallization.
9. Store the paraffin blocks in the freezer for a few hours or until sectioning is performed.

LAB REPORT FORMAT:

NAME OF THE GROUP MEMBERS:

DATE OF SUBMISSION:
(deadline: March 18)

PH 122 LAB EXERCISE: TISSUE PROCESSING

I. FIXATION
Specimen

Time Obtained

Time Fixed in Formalin

Length of Fixation

Color

Texture
(smsooth/ rough)

Consistency
(Hard/Soft)

1.
2.
3.
4.
5.
After Fixation
Specimen
1.
2.
3.
4.
5.
(Include images here, properly labelled with name of specimen)
II. Dehydration
After Dehydration
Specimen

Length of
Dehydration

Color

Texture
(smsooth/ rough)

Consistency
(Hard/Soft)

1.
2.
3.
4.
5.
(Include images here, properly labelled with name of specimen)
III. Clearing
After Clearing
Specimen
1.
2.
3.
4.

Color

Texture
(smsooth/ rough)

Consistency
(Hard/Soft)

Image

5.
IV. Embedding
(image of paraffin block)
V. Sectioning
(image of sections on slide)
VI. Staining
(image of section after deparaffinization, nuclear staining, decolorization, blueing, counterstaining,
mounting, and final appearance of section)
VII. Microscopic examination
(image of each tissue in LPO and HPO)
APPENDIX
(task of each group member)