SCHOOL OF MEDICAL

LABORATORY SCIENCE
HISTOPATHOLOGIC AND CYTOLOGIC TECHNIQUES (LEC.)
SAN PEDRO COLLEGE – MAIN
CAMPUS
Instructor’s Name: Ma’am. Doren Venus Otod, RMT
AY 2022 – 2023 - 2ND SEMESTER WEEK 03 - LESSON 05 BIOPSY AND POST MORTEM EXAMINATION

1. PATHOLOGIST (Head of the Laboratory)

OUTLINE → Sections large and hollow organs to allow fixation
I. Overview of the → examines the tissue sections, cytologic slides under the
II. Examination of Tissues microscope for diagnosis
Histopathology Laboratory
a. Autopsy 2. ASSOCIATE PATHOLOGIST
a. Professionals under
b. Biopsy → assistant of the pathologist
Histopathology
i. Fresh Tissue → monitor staff performance
b. Quality in
Examination → pinpoint problematic situations and find solutions
Histopathology
ii. Fixed Tissue 3. HISTOTECHNOLOGIST/HISTOTECHNICIAN
Laboratory
Examination → assist the pathologists by providing spx slides that are
c. Inside the Laboratory
properly labeled, processed, stained, and mounted
→ ensures high quality conditions of equipment, formalin, and
LEARNING OBJECTIVES other chemicals and reagents
● Know the individuals behind histopathology laboratory, as well → work systematically to minimize errors
as the various processes performed inside the laboratory. → analyze problems and corrects them
● compare the different principles associated in biopsy and
autopsy. B. QUALITY IN HISTOPATHOLOGY LABORATORY
● Identify the instruments used in routine and automated tissue ● QUALITY ASSURANCE
processing in the laboratory, as well as their functions. → ensuring that everything is right (test, time, specimen,
● discuss proper storage conditions, borrowing policies, disposal patient, diagnosis and price)
proceedings, and retention time of specimens, blocks, slides, → we cannot do repeat collection
and reports. ● QUALITY MANAGEMENT SYSTEMS
→ set of coordinated activities to regulate a lab in order to
I. HISTOPATHOLOGY LABORATORY continually improve its performance
→ considers pre-analytic (receiving to specimen mounting) ,
analytic (actual reading of the slides) and post-analytic phase
(releasing of the results)

Figure 2. Workflow in the Laboratory

Figure 1. Sections in the Laboratory
HISTOPATHOLOGY
● The study of tissues affected by disease C. INSIDE THE LABORATORY
● Useful in making a diagnosis and in determining the severity
and progress of a condition
● diagnose the changes happening in the cells

HISTOPATHOLOGIC TECHNIQUES
● includes all activities done in the laboratory in order to
produce a suitable specimen slide for viewing by the
pathologist
● 5 major processes performed in the lab: Tissue
processing, cytology, frozen biopsy, special staining,
immunohistochemistry/ immune staining

A. PROFESSIONALS UNDER HISTOPATHOLOGY - Patient report is the result

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 SCHOOL OF MEDICAL
LABORATORY SCIENCE
HISTOPATHOLOGIC AND CYTOLOGIC TECHNIQUES (LEC.)
SAN PEDRO COLLEGE – MAIN
CAMPUS
Instructor’s Name: Ma’am. Doren Venus Otod, RMT
AY 2022 – 2023 - 2ND SEMESTER WEEK 03 - LESSON 05 BIOPSY AND POST MORTEM EXAMINATION

- Example of a telephone report is during frozen section, - the problem with this is that frequently, the
when a pathologist relays the information to the surgeon organ that we choose to examine is not the
- Preliminary report is the status of the sample, 48-72 hrs cause of the death
from receiving the specimen ➢ Complete - examination of the entire body (both
- Final report is the report of the pathologist upon completing external and internal)
and reading the sample under the slide
- Incident report are the documents prepared if there are 3. Manner of Incision
problems inside the laboratory ➢ Y-shaped - pathologist will begin at the chest and
- Instruments in the lab are used for the different processes abdomen autopsy by making a y-shaped incision
that we are performing - the arm of the “Y” starts from each of the
shoulder and meet at the midchest
● TYPES OF RESULTS - the stem of the “Y” will run down to the pubic
→ surgical pathology (biopsy) region
→ cytopathology (cytological tests) ➢ Straight cut - “I”; starts from the symphysis menti down
→ autopsy report to the pubis region
- the disadvantage is that, the marking or the
● Turnaround Time (TAT) of Results stitchings will be prominent in front of the neck
→ surgical pathology and cytology = 2 days and the thorax
→ Frozen Sections = 5-15 mins - the back of the neck cannot be properly be
→ Autopsy Report = 7 days visualized
→ Note: Time will vary depending on the protocol of the
laboratory but these are the recommended. ● Different techniques performed in Autopsy

II. EXAMINATION OF TISSUES

● AUTOPSY VIRCHOW TECHNIQUE

→ Post mortem (somatic death) examination → most widely used technique
→ from the Greek words “Autopsia Cadavernum” → organs are removed one at a time
● BIOPSY ROKITANSKY TECHNIQUE
→ Ante Mortem examination → the organ is not removed from the body during dissection
→ “Bio” Life; “Opsia” To See followed by the removal of the organ bloc
GHON
→ removal of the region or a cavity of the same group
A. AUTOPSY → ex. Thoracic organs will be removed; grouped according to
→ aka Necropsy; Thanatopsy their region
→ Post-mortem examination of tissues LETULLE
→ Purposes: → entire organs are removed from the body
➢ Determine cause of death and extent of injury → its disadvantage is that since the organs are too heavy and
➢ Uncovering existence of an undetected disease big, it cannot fit on the standard chopping boards used

● Types of Autopsy according to:
1. Purpose:
➢ Medical/Hospital - performed on patients who died in
the hospital during course of treatment
➢ Medico-legal - for criminal trials; suspicious death
2. Completeness:
➢ Partial - examination of a specific area of a body
Note: Autopsy requires the following documents:
● Written or informed consent from the legal next-of-kin
- the order of priority: if dead body is married, the
spouse is the next of kin; if not present, the adult
child; either of the parents of the cadaver; adult
sibling; grandparents; guardians
● medical abstract or clinical data
● Autopsy request (suspicious evidence of foul play)
- police reports

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 SCHOOL OF MEDICAL
LABORATORY SCIENCE
HISTOPATHOLOGIC AND CYTOLOGIC TECHNIQUES (LEC.)
SAN PEDRO COLLEGE – MAIN
CAMPUS
Instructor’s Name: Ma’am. Doren Venus Otod, RMT
AY 2022 – 2023 - 2ND SEMESTER WEEK 03 - LESSON 05 BIOPSY AND POST MORTEM EXAMINATION

4. Lesions that are encountered during dissection should be

obtained early and placed in fixative before the organ is
fully incised.

B. BIOPSY

Figure 6. Autopsy personnels.

● Personnel involved in the process of Autopsy:

→ Coroner - a public official who is empowered to order an
inquest into the manner or cause of death.
▪ Without the request of the coroner, an autopsy cannot
continue.
→ Prosector - pathologist who performs the dissection
→ Diener - comes from the German word “leichendiener”
meaning “servant of the dead”: assists during autopsy, and
assumes many and varied responsibilities in the autopsy
laboratory.

Note: Personnel must wear full personal protective equipment

(PPE) when performing this type of procedure. Figure 8. Types of Biopsy.

● Ante-mortem examination of tissues

→ “ante” = before; “mortem” = death
→ Removing a piece of tissue or sample of cells from the body
that will be analyzed in the laboratory.
● Examination of tissue sample from the living

Figure 7. Example of enameled pots.

● Proper handling of Autopsy specimens

1. Organ blocks removed from the body cavity should be
thoroughly washed of blood using cool or cold water to
minimize the blood staining of organs. Never use hot
water.
2. Organ blocks are placed in a large enameled pot Figure 9. FNA (up); Core needle (down).
containing fixatives.
3. Tissue should not be pressed against each other or the ● Types of Biopsy
bottom or walls of the container. 1. Fine needle aspiration (FNA)
➢ The container should be big enough for the sample ➢ Simplest, least invasive.
itself plus the fixative. ➢ Uses a very thin needle attached to a syringe to take
out a small amount of fluid and tissue from the area.

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 SCHOOL OF MEDICAL
LABORATORY SCIENCE
HISTOPATHOLOGIC AND CYTOLOGIC TECHNIQUES (LEC.)
SAN PEDRO COLLEGE – MAIN
CAMPUS
Instructor’s Name: Ma’am. Doren Venus Otod, RMT
AY 2022 – 2023 - 2ND SEMESTER WEEK 03 - LESSON 05 BIOPSY AND POST MORTEM EXAMINATION

➢ According to the image, a small amount of fluid and 5. Punch

tissue will be taken from the area. It is usually guided by ➢ For skin; uses circular blade to obtain deeper skin
CT scans for the doctor to know where the nodule is sample that removes a short cylindrical core of tissue
placed. (“apple core”).
➢ Its problem is that the sample obtained is usually ➢ The size of the circular blade is usually 3-4 millimeter in
inadequate. diameter.
2. Core needle 6. Shave
➢ Uses slightly larger needle ➢ For skin; small fragments of outer layers of skin are
➢ Removes small column of tissue (1/16 inch in diameter, “shaved” or scraped.
1/2 in long) 7. Curettage
➢ Does not only remove the cells but also a small amount ➢ Tissues are removed from body cavity (canals) using a
of surrounding tissue. curette.
➢ The needles are bigger and have a notch at the tip of ➢ The tip of the curette is like a small scoop or hook.
the needle. Hence, a small amount of tissue can be
taken.

Figure 12. Examination methods diagram.

● If the sample is placed in a container with a fixative or formalin,

it means it is for fixed preparations.
Figure 10. Incisional and excisional biopsy. ● If the sample is received in a fresh state, depending on the
request of the doctor, the following procedures will be
3. Incisional
performed.
➢ Surgical; small part of a large lesion or tumor is taken.
➢ The doctor will slice into the lesion and will identify if
further surgery is needed or the entire lesion must be I. FRESH TISSUE EXAMINATION
removed.
4. Excisional
➢ Surgical; the entire affected area is taken.

Figure 13. Teasing (left) and crushing (right).

● Teasing/Dissociation
→ Pressing the samples for it to be flattened and placed on a
slide. Supravital stains can be used during this process.
● Crush Prep/Squash Prep
→ The diameter of the specimen must not be more than 1
millimeter.
→ Compressing two slides or cover slips. Use supravital stains
Figure 11. Punch (top), shave (middle), curettage (bottom). for this process.

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 SCHOOL OF MEDICAL
LABORATORY SCIENCE
HISTOPATHOLOGIC AND CYTOLOGIC TECHNIQUES (LEC.)
SAN PEDRO COLLEGE – MAIN
CAMPUS
Instructor’s Name: Ma’am. Doren Venus Otod, RMT
AY 2022 – 2023 - 2ND SEMESTER WEEK 03 - LESSON 05 BIOPSY AND POST MORTEM EXAMINATION

● Smear Preparations

Figure 14. Streaking (top) and spreading (bottom). Figure 16. Cryostat.

→ Streaking ● Frozen Sections

▪ Getting a sample and making a smear using a zigzag → In 10-15 minutes, the results will be released for this type of
manner. testing.
→ Spreading → Recommended for rapid diagnosis of tissue. and
▪ Getting a small amount of sediment and placing it on a → Requested during intra-operative procedures.to help the
slide not too thin nor too thick. surgeon in choosing his next plan of action.
▪ Intraoperative means there is an ongoing operation and
the sample will be sent to the laboratory for the surgeon to
check if the entire organ will be removed. If it is not
cancerous, the operation will not be pushed through.
→ Fresh tissues are frozen using a cryostat or freezing
microtome.
→ The temperature should be maintained at -10 to -30 degrees
celsius.

II. FIXED TISSUE EXAMINATION

The end goal is to produce a tissue section of good quality that
allows for adequate interpretation of microscopic cellular changes
(for diagnosis).

Accomplished by fixing the tissues and carefully processing them

to preserve their structures, then impregnating them with
hardening substance to permit making thin slices suitable for
Figure 15. Pull-apart (top) and touch prep (bottom).
staining and microscopic evaluation.
→ Pull-apart
● Following processes that a fixed tissue undergoes:
▪ Using two slides, one wrap will be placed on one end of
1. Fixation
the slide. Another slide will be placed on top, and then the
➢ Preserving the tissue constituents in a lifelike manner
slide will be pulled in an opposite direction with single
as possible.
uninterrupted movement.
2. Decalcification
→ Touch Prep/Impression Smear
➢ Optional. Not processed for all types of tissue. Only
▪ Freshly cut tissue will be pressed on a clean slide allowing
done for hard tissues such as bone and cartilage.
cells to be transferred directly on the slide. After that a
3. Dehydration
cover slip will be placed and observed under the
➢ Process of removing the water that was accumulated on
microscope.
our tissue.
4. Clearing/Alcoholization
Note: During smear preparations, all samples must be centrifuged
➢ Removing the dehydrating agent that was used during
before actual processing. Sediments are used, while the
the dehydration process.
supernatant is discarded.
5. Infiltration/Impregnation
➢ Replacing the clearing material during the clearing
process with impregnating material.
➢ Impregnating material gives firm consistency to the
tissue for ease of handling and sectioning. Usually it is
the paraffin wax.

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 SCHOOL OF MEDICAL
LABORATORY SCIENCE
HISTOPATHOLOGIC AND CYTOLOGIC TECHNIQUES (LEC.)
SAN PEDRO COLLEGE – MAIN
CAMPUS
Instructor’s Name: Ma’am. Doren Venus Otod, RMT
AY 2022 – 2023 - 2ND SEMESTER WEEK 03 - LESSON 05 BIOPSY AND POST MORTEM EXAMINATION

6. Embedding/Casting/Molding
➢ Placing the tissue into the mold with molten wax to form
a solid block tissue.
7. Blocking
8. Trimming
➢ Removing excess paraffin wax that was accumulated
during the embedding process.
9. Sectioning/Microtomy
➢ Using a microtome, the tissue will be cut into fine tissue
sections to create tissue ribbon.
10. Staining
➢ Applying dyes onto the sections.
➢ Most common dye in histopath for fixed tissue
examination is hematoxylin and eosin stain.
11. Mounting
➢ Facing mounting media and covering it with a slip to
protect the sample and allow storage of the slides.
12. Labelling
➢ Follow the accession number of the patient
13. Microscopic Examination

REFERENCES

● Kumar, V., Abbas, A. K., & Aster J. C. (2015). Robbins and

Cotran Pathologic Basis of Disease (Tenth Edition).
Philadelphia, PA: Elsevier/Saunders
● Shedge, R., Krishan, K., Warrier, V., & Kanchan, T. (2019).
Postmortem Changes. In StartPearls [Internet]. StartPearls
Publishing

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