Histopathology

PROPER DONNING Biopsy Specimens

1. Inner gloves - Is a sub group of surgical specimens, however, these small
2. Gown tissue samples are obtained in procedures other than
3. Mask surgery.
4. Headcap
5. Facemask/ goggles Needle Aspiration Biopsy
6. Shoe cover - A biopsy obtained by inserting a hollow needle through the
7. Outer gloves skin into an internal organ.

Excision Biopsy
PROPER DOFFING - This involves the removal of a small lesions
1. Outer gloves
2. Shoe cover
Endoscopic Biopsy
3. Facemask/ googles
- A small specimen of digestive or respiratory tract obtained
4. Gown
by inserting an instrument periorally or perianally and
5. Headcap
removing a piece of tissue.
6. Mask
7. Innergloves
Wedge Biopsy
- This term describes a small pie shaped piece of a particular
ANATOMICAL PATHOLOGY organ obtained during exploratory surgery.
Sections:
 Surgical Abrasion Biopsy
 Cytology - A specimen received that is abraded from the surface of the
 FNAB/ CNAB skin or mouth lesion done using a sponge or brush.
 Slide Review
 Breast Panel Cone Biopsy
 Frozen Section - Special procedure for collection of specimen in which a
 Surepath/ PAP smear cone shaped portion of the cervix is removed.

10 STEPS OF TISSUE PROCESSING GROSS EXAMINATION AND FIXATION

1. Fixation
2. Dehydration Gross examination
3. Clearing - assessment of tissue specimens by the unaided eye
4. Impregnation
5. Embedding Fixation
6. Trimming - The first and most critical step of tissue processing
7. Smearing
8. Staining Pathologist
9. Mounting - Gross examination is done by who
10. Labelling
Gross description and tissue block
5 LITERS 10% NBF (Neutrally Buffered Formalin) - The 2 end products of gross examination
1. 20g monobasic powder
2. 32.5g Dibasic powder Brain
3. 500mL 37% Formaldehyde - Tissues can be cut & trimmed without prior fixation except
4. 4.5L Distilled H2O for the?

GROSSING TISSUE SPECIMEN Photographs

- Provides a better 3-Dimensional understanding of the
Gross specimen
- A description and dissection of tissue specimen, done after
tissues are fixed or partially fixed. Millimeters
- A step that preceded tissue processing, embedding, - Unit for very small specimen in grossing
microtomy and staining.
Centimeter
Types of Specimen - The standard unit of measurement
 Surgical specimens
 Autopsy specimens 2-3mm
- Standard tissue thickness in gross examination
Surgical Specimens
- Specimens obtained during operative surgery Nitric Acid
- Discrete areas of calcification is decalcificied using this acid

Angela Obrique Veraye

 Histopathology

Bread Loafing
- Knife technique for a live specimen. Non coagulant
- Create a gel that makes it difficult to penetrate by
1:20 subsequent solutions
- Ratio of tissue to fixative for routine exam
pH 6-8
1:50 - The satisfactory pH level of a fixative
- Ratio of tissue to fixative in museums
-60 °C
Fixation - The temperature required for rapid fixation
- A complex series of chemical events which bring about
changes in the cell-like hardening 400-450 mOsm
- The best result obtained in fixation is attained with—?
Crosslinking of proteins
- The ________ produces denaturation of coagulation of Mordant
proteins so that the semifluid state is converted into - Promotes staining or inhibit certain dyes in favor of another
semisolid state
Simple fixative
Primary aim - A fixative made up of only 1 component
- To preserve the morphologic and chemical integrity of the
cell in a life-like manner as possible Cytologic fixative
- A specific part and particular microscopic element of the
Secondary aim cell itself
- To harden and protect the tissue from trauma of further
handling 10% buffered formalin
- Most widely used fixative for routine histology
Autolysis
- A decomposition of a certain material due to hydrolytic 10% Formal Saline
enzymes - A simple microanatomical made up of 40% saturated
formaldehyde diluted to 10% sodium chloride (pH= 6.8)
Lysozome
- An example of a hydrolytic enzyme Alcoholic formalin
- Used for primary fixation specimens and for post fixation of
Putrefaction large fatty specimens
- The decomposition of a substance due to bacterial/fungal
colonization Glutaraldehyde
- Made up of two formaldehyde residues, linked by a three
Solidification carbon chain. It also doesn't cause dermatitis
- Converts the normal semifluid consistency of cells to an
irreversible semisolid consistency Carnoy’s fixative
- The fastest fixative
Optical Differentiation
- Alters the refractive indices to varying degrees so that Methanol 100%
unstained components are more easily visualized than - Excellent for fixing dry and wet smears, blood smears, and
when unfixed bone marrow

Formaldehyde ISOPROPYL ALCOHOL 95%

- This fixative intensifies the staining character of - Used for fixing touch preparations
hematoxylin

37-56°C ETHANOL
- What is the suggested heat temperature to hasten fixation - Used as a simple fixative and is incorporated into compound
fixatives for better results
Non additive
- Stabilizes the tissue by removing the bound water attached
to the hydrogen bond of a certain group within the protein CARNOY'S FIXATIVE
molecule - Permits good nuclear staining and differentiation

Additive
- Fixative is taken in and becomes a part of the tissue NEWCOMER'S FLUID
- Recommended for fixing mucopolysaccharide and nuclear
Coagulant proteins
- Creating a network that allows solutions to readily
penetrate the interior of the tissue

Angela Obrique Veraye

 Histopathology

METALLIC FIXATIVES POST-CHROMATIZATION

- Fixes tissues through an unknown mechanism which - A form of secondary fixation whereby a primarily fixed
increases the staining brightness and gives excellent tissue is placed in an aqueous solution of 2.5-3% potassium
nuclear detail of our specimen dichromate for 24hrs

ZENKER'S SOLUTION
- Permits brilliant staining of nuclear and connective tissue WASHING OUT
fibers - the process of removing excess from the tissue after
fixation in order to improve staining and remove artifacts
from the tissue
HEIDENHAIN'S SUSA SOLUTION
- Permits most staining procedures to be done CADMIUM/COBALT SALT
- Prevents fat dispersal

FLEMMING'S SOLUTION
- Permanently fixes fat 3 MONTHS
- To prevent bleaching, change the fixative every _____

REGAUD'S
- Recommended for preservation of chromatin, mitochondria, 70%
mitotic figures, golgi bodies, RBC, and colloid-containing - To restore natural specimen color, what is the percentage
tissues of the alcohol solution that the specimen should be
immersed in?

ORTH'S FLUID
- Demonstrates rickettsiae and other bacteria MAGNESSIUM/CALCIUM CARBONATE
- Removes formic acid formation
3% POTASSIUM DICHROMATE
- Fixes but does not precipitate cytoplasmic structures,
preserves lipids and mitochondria BLEACHING, DISPERSAL OF FAT, DISSOLUTION
- Effects of prolonged fixation

PICRIC ACID
- Stains yellow, used for strong saturated aqueous solutions

LITHIUM CARBONATE
- Picric Acid's yellow stain may be removed by another acid
dye or ______

BOUIN'S SOLUTION
- Allows brilliant staining with trichrome method

BRASIL'S SOLUTION
- Better and less "messy" than Bouin's solution

4% LEAD ACETATE
- Takes up CO2 to form insoluble lead carbonate

ACETONE
- Used in fixing brain tissues (rabies)

SECONDARY FIXATIVE
- The process of placing an already fixed tissue in a second
fixative

Angela Obrique Veraye