- Bilateral organs o Examples: Extremities, kidneys, lungs and ovaries) - Miscellaneous o Examples: age, sex, ward

MLSP: HISTOPATHOLOGY SECTION Flashcards by Zelene Ignacio

involves different procedures that have been adopted for the preparation of materials and tissue for microscopic examination

Histopathologic Techniques

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12 steps in Histopathologic Techniques:

Numbering
Fixation
Dehydration
Clearing
Wax impregnation
Embedding
Blocking
Trimming
Sectioning
Staining
Mounting
Labelling

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(t or f)

o There is a 13th step which is decalcification (done or placed in between fixation and dehydration; for
bones including teeth)
o All in all, it will take 24-48 hours to do

true

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• Validates if the specimen is adequate or good for
tissue processing
• 1st person that will receive the specimen and put it
into the container

RECEPTIONIST

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Proportional to the size of the specimen [larger
than the organ; the organ or specimen should
be fully submerged]

Container: contain fixatives

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(t or f)
Container: contain fixatives

o If the specimen is the lungs, wrap it in gauze pads to ensure that the air inside of it wouldn’t let the organ float, hindering it to submerged fully to the
fixative agent

true

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Container: contain fixatives

[to see the organ]

clear

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Container: contain fixatives

[no leaking]

Unbreakable

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Container: contain fixatives

[easy access]

Wide mouthed bottle

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(t or f)

• The receptionist must not accept specimens that
are only placed in plastic
• Medical Technologists assist pathologists in
autopsies.

true

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(t or f)

Portion of the specimen will be placed in tissue
cassettes and the rest of the organ will be
contained
o Its shelf life is 1 year or so depending on
the policy of the hospital

true

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• Specimen Source:

Bilateral organs
o Examples: Extremities, kidneys, lungs
and ovaries)
Miscellaneous
o Examples: age, sex, ward

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• Basic information needed:
1. Date and time
2. Name of the patient
3. Specimen Number
- C- Cytology specimen
- A- Anatomical specimen
- S- Surgical specimen
o Example: S-09-2111

NUMBERING

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Most critical step in histopathological techniques
- If you mess this step, you will mess up everything

FIXATION

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FIXATION

preserve the morphology and chemical constituents of the tissue [tissue should be alive as possible]

Primary aim

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FIXATION

protect and harden the specimen for further handling

Secondary aim

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TYPES OF FIXATIVES

ACCORDING TO ACTIONS

Microanatomic Fixative
Cytological Fixative
Histochemical Fixative

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TYPES OF FIXATIVES

ACCORDING TO COMPOSITION

Simple Fixative
Compound Fixative

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Involves small tissue or organ
- 10% Formol Saline
- 10% Neutral Buffered Formalin [tinitimpla]

Microanatomic Fixative

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Involves body fluid or secretion
- Nuclear fixatives: Flemming’s Fluid, Bouin’s Fluid,
Heidehain’s Susa
- Cytopplasmic Fixatives: Kelly’s Fluid, Orth’s Fluid

Cytological Fixative

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Involves tissue containing labile substances
- 10% Fomol Saline
- Absolute Ethyl Alcohol
- Acetone

Histochemical Fixative

Uses only on chemical for
fixation
- Aldehydes: Formaldehyde, Glutaraldehyde
- Metallic Fixative: Mercuric Chloride, Chromate Fixatives, Lead Fixatives

Simple Fixative

The use of two or more chemicals for fixation

Compound Fixative

• Utilizing chemical known as

dehydrating agents

• Removing of intracellular and extracellular water and fixatives in the tissue

DEHYDRATION

DEHYDRATION examples:

-Alcohol - Acetone - Dioxane - Tetrahydrofuran - Cellosolve (Ethylene glycol monoethyl ether)

• Removing of dehydrating agents • Xylene – most commonly used [organic solvents]

CLEARNING

Also known as INFILTRATION OR WAX IMPREGNATION

IMPREGNATION

• The process whereby the clearing agent is completely removed from the tissue and replace by a medium that will completely fill all the tissue cavities.

IMPREGNATION

the simplest, most common and best embedding medium used for routine tissue processing - Surface area of the tissue is larger; clearing agent is removed; specimen is further hardene

PARAFFIN WAX IMPREGNATION

Also known as CASTING OR BLOCKING

EMBEDDING

• The process by which the impregnated tissue is placed into a precisely arrange position in a mold containing medium which is then allowed to solidify o Larger surface area of wax or tissue block; larger piece to hold

EMBEDDING

• FOUR TYPES OF TISSUE IMPREGNATION:

- Parrafin wax - Celloidin - Gelatin - Plastic

• Allows the medium to solidify to produce tissue block [inaayos yung shape]

BLOCKING

• Process of removing excess wax after embedding • Can use knife/blade or heated spatula

TRIMMING

• Also known as CUTTING OR MICROTOMY • The process by which processed tissue is cut into uniformly thin slices to facilitate studies under microscope o Making tissue ribbons [very thin segments of wax and tissue]

SECTIONING

machine or instrument used for cutting sections of tissue - Blade and holder - Thin slices; cannot do manually - In this step, you will see the flaws and mistakes you’ve made throughout the process

MICROTOME

KINDS OF MICROTOME - Simplest and oldest type of microtome

Rocking Microtome

KINDS OF MICROTOME - for cutting serial sections of tissue specimen [common]

Rotary Microtome

KINDS OF MICROTOME - Most dangerous type of microtome

Sliding Microtome

KINDS OF MICROTOME - For urgent surgical biopsies specimen

Freezing Microtome

KINDS OF MICROTOME - Permits rapid penetration of tissue biopsies for surgical pathology - Also known as COLD MICROTOME

Rotary Microtome

KINDS OF MICROTOME - Specimen for electron microscope

Ultrathin Microtome

• Tissue constituent are demonstrated in sections by direct interaction with dye or staining solution producing coloration of the active tissue component

STAINING

- Utilizes micro-anatomical studies of tissue - It is a regressive staining method - Haematoxylin is used to stain nuclear components - Eosin is used for cytoplasmic components - Can be used as a pair

HAEMATOXYLIN AND EOSIN STAINING

the solution in which the specimen is embedded, generally under a cover glass. o Adhesive agent to protect the specimen

MOUNTING MEDIUM

• It may be liquid, gum or resinous, soluble in water, alcohol or other solvents and be sealed from the external atmosphere by non- soluble ringing media o Sealed to ensure that no dust or insect can pass through the slide o Indefinite shelf life

MOUNTING

• Date and time • Name of the patient • Specimen Number - C- Cytology specimen - A- Anatomical specimen - S- Surgical specimen o Example: S-09-2111

LABELLING

SPECIMENS FOR EXAMINATION • Performed regularly even in pregnant women without undue risk • EXAMPLE: Vaginal smear

GYNECOLOGICAL SPECIMEN

SPECIMENS FOR EXAMINATION • EXAMPLE: Respiratory Tract specimens: - Sputum - Bronchoalveolar lavage (BAL)

NON-GYNECOLOGICAL SPECIMEN

SPECIMENS FOR EXAMINATION • Determine the presence of urethral cancer - Pattern called “ferning”

URINE