Ph 122 Lab Exercise
3
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 3h 95 % Alcohol 1 h 95% Alcohol 1 h Absolute Alcohol I 1 h (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 1 h Clearing Xylene I 1 h (if xylene becomes turbid upon immersion of tissues, return the tissues in absolute alcohol and transfer to xylene after an hour) Xylene II 1 h Infiltration Paraffin I 1 h Paraffin II 1 h Embedding 4. Make a paper boat by cutting a 12cm x 9c m 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 f ilm of paraffin at the bottom to partially solidify. This will only take a few seconds. 6. Lift the tissue from the final wax, with previ ously 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 ¾ 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 f ew hours or until sectioning is performed.
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Transcript of Ph 122 Lab Exercise
8/10/2019 Ph 122 Lab Exercise
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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 3h
95 % Alcohol 1 h
95% Alcohol 1 h
Absolute Alcohol I 1 h
(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 1 h
Clearing
Xylene I 1 h
(if xylene becomes turbid upon immersion of
tissues, return the tissues in absolute alcohol
and transfer to xylene after an hour)
Xylene II 1 hInfiltration
Paraffin I 1 h
Paraffin II 1 h
Embedding
4. Make a paper boat by cutting a 12cm x 9cm glossy paper and folding it to make a 9cm3. 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 themold. 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 ¾ 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.
8/10/2019 Ph 122 Lab Exercise
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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
1.
2.
3.
4.
5.
After Fixation
Specimen Color Texture
(smsooth/ rough)
Consistency
(Hard/Soft)
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 Color Texture
(smsooth/ rough)
Consistency
(Hard/Soft)
Image
1.
2.
3.
4.
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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)
