Enclonar, Kimberly / MLS 3A

• Hazards should be properly labeled:

Risk Mgt & Gross Exam o Chemical (mixture: with ingredients) &
October 19, 2020 manufacturers name
Gabriel Ponce de Leon, RMT, AMT, ASCPi, MS o Date of purchase and expiration date
o Hazard warnings and safety procedure
Histopathology
• Documentation and record-keeping
• Branch of pathology which deals with the study of
disease in a tissue section.
Types of Hazards
• A series of steps before it reaches the pathologist's
• Chemical
desk to be meticulously examined thru a microscope
• Physical
depending upon the specimen type.
• Biological
• Acceptable slide: the tissue must be prepared in such
manner that it is sufficiently thick or thin to be
Chemical Hazard
examined microscopically and all the structures in a
• Cleaning agents and disinfectants, anesthetic gases,
tissue may be differentiated.
solvents, etc.
• Biopsy - Small piece of lesions or tumor which in set
• Permissible Exposure Limits (PELs), Threshold Limit
for diagnosis before final removal of the lesion or the
Values (TLVs), Occupation Exposure Limits (OELs) -
tumor (incisional biopsy)
maximum allowable airborne concentration.
• One of the most dangerous sections in the laboratory.
• Types of chemicals:
• Biological, chemical and physical hazards. o Irritants - cause reversible inflammatory effects.
• Common hazards: o Corrosive chemicals - cause destruction or
o Electrical equipment, combustible material,
irreversible alterations.
sharp objects, chemical reagent, human tissue,
o Sensitizers - cause allergic reactions.
radiation exposure
o Carcinogens - substances that induce tumors.
o Toxic materials - capable of causing death by:
Risk Management ▪ Ingestion
• Process of ensuring and maintaining personal as well ▪ Skin contact
as environmental health and safety in the laboratory. ▪ Inhalation
• Minimize risks associated with day-to-day activity by • Never add water to acid!
using safety guards and checking the quality of • Cryogens
reagents. o Produce substance with temperatures below -
• First step: identify all electrical, mechanical and 153C (-243F) and a boiling point of -196C (-
biological hazards. 321F).
o Identification health hazards o Solid CO2 or dry ice is also often used -78C (-
• Standard Operating Procedures (SOPs) must be 109F)
detailed to include control of hazardous substances, ▪ To dispose, sublimate or evaporate.
risk assessments, and other health and safety
information relevant to handling of specimens. Physical Hazards
• Work Practice controls: good housekeeping, • Most obvious are slips and falls, and ergonomic
ergonomic arrangement, decontamination with hazards.
bleach, capping of flammable reagents, mouth • Combustibles: substances that ignite at
pipetting prohibition o Flash point
• One of most common accidents - cut from microtome ▪ OSHA - 100F (38C)
knives ▪ DOT - 141F (60.5C)
• Universal precaution is the most effective way of • Flammables: Substances having flashpoint below
preventing infection 100F or 38C
• Explosives: oxidizers, may initiate combustion
How to prevent? • Electrical Hazards
• Personal Hygiene
• Practice hand washing Waste Disposal Color Coding
• Avoid food and drinks in refrigerator
Yellow Infectious
• Complete PPE
• Closed shoes Black General Waste, non-infectious DRY
• Hair must be fixed Green General waste, non infectious WET

Risk Management Process Orange Radioactive

• Assessing the risks
• Prioritize your risks Terms to Remember
• Figuring out your risk profile OSHA Occupational Safety and Health Administration
• Choosing your risk strategies
• Execute risk strategies MSDS Material Safety Data Sheet
• Measure residual risks STEL Short Term Exposure Limit
PEL Permissible Exposure Limit
General Precautions
• Volatile substances must be handled under the fume TLV Threshold limit values
hood. OEL Occupational Exposure Limit
• Tissues collected should be stored in formalin and
may be disposed by incineration or by putting them
through a "tissue grinder"
• Warning signage: must be posted in the laboratory
 Enclonar, Kimberly / MLS 3A
Biological Hazards Methods of Fresh Tissue Examination
• Anything that can cause disease in humans 1. Teasing or Dissociation
• Allergens - one of the most important health hazards o Process whereby a selected tissue specimen is
• Complete penetration by alcohol will destroy all immersed in isotonic salt solution such as
infectious agents except prions. normal saline or Ringer's solution.
o Prions can cause spongiform encephalopathies o Dissection - pin
such as Creutzfeld-Jakob disease (CJD), scraple, o Mounted as wet preparation, either stained
and mad cow disease. (supravital stain) or unstained (Phase Contrast &
Brightfield)
Handling Spills o Not permanent
• Small spills - can be safely handled by immediate 2. Squash Preparation (Crushing)
staff. o Small piece of tissue (not >1mm in diameter) is
• Use of 10% bleach solution. place on a slide and forcibly compressed with
Disposable plastic aprons Chemical spills another slide or a cover glass.
o Supravital (vital) stain - place in junction of the
Disposable gowns Biohazards slide and absorbed through capillary action.
Sodium hypochlorite Biohazards 3. Smear Preparation
o Sections or sediments are spread lightly over a
Baking soda Acids
slide using a wire loop or applicator stick.
Vinegar (5% acetic acid) Alkalis o Useful: cytological examinations (cancer
diagnosis)
Examination of Fresh Tissue o Types:
• Histology - Microscopic study of normal tissues of the ▪ Streaking
body. □ Using an applicator stick or a platinum
• Histopathology - study of tissues affect by disease. loop, the material is rapidly and gently
• Histologic or histopathologic techniques - preparation applied in a direct or zigzag line.
of material for such studies. ▪ Spreading
□ A selected portion of the material is
Factors for Evaluation transferred to a clean slide and gently
• Structural & chemical components of the cells to be spread into moderately thick film by
studied teasing the mucous strands apart with
• Nature and amount of the tissue to be evaluated an applicator stick.
• Need for an immediate examination of tissue □ Maintains cellular interrelationships.
structure □ Recommended for: fresh sputum,
• Not part of biopsy bronchial aspirates, and thick mucoid
secretions.
Surgical Procedures ▪ Pull-apart
• Fine needle aspiration □ Done by placing a drop of secretion or
o Simplest, least invasive test sediment upon one slide and facing it
o Uses the smallest needle to simple remove cells with another slide. Material disperses
from the area of abnormality. evenly over the surface of the two
• Core Needle Biopsy slides. The two slides are then pulled
o Removes not only cells, but a small amount of apart with a single uninterrupted
the surrounding tissue. motion.
o Provides additional information to assist in the □ Used for thick secretions such as
examination of lesions. serous fluids, concentrated sputum,
• Incisional Biopsy enzymatic lavage (GI tract) and blood
o Takes out even more surrounding tissue. smears.
• Excisional Biopsy 4. Touch Preparation (Impression Smear)
o Removes the entire area in question. o Special method of smear preparation whereby
• Punch biopsy the surface of a freshly cut piece of tissue is
o Primary technique for obtaining diagnostic full- brought into contact and pressed on to the
thickness skin specimens. surface of a clean glass slide, allowing cells to be
o Uses circular bladed yielding 3-4 mm cylindrical transferred directly to the slide for examination
core of tissue sample. by Phase Contrast Microscopy.
o Cells may be examined without destroying their
• Shave biopsy
o Small fragments of tissue are "shaved" from a intercellular relationship.
surface, usually skin. 5. Frozen Section
o Most ideal and preferred.
• Curettings
o Tissue is scooped or spooned to remove tissue o Rapid diagnosis of a pathologic tissue.
o Recommended for: lipids and nervous tissue
or growths from body cavity such as
endometrium or cervical canal. elements.
o Done on muscle and nerve biopsies, as well as
Autolysis tumors.
o Slices: very thin, 10-15u is cut on a microtome
• Once tissues are removed from the body, their
proteins and cells are digested and broken down by with CO2 or in a cryostat
o Cryostat: cold chamber kept at an atmospheric
their own enzymes, independent of a bacterial action.
temperature of -10 t -20C.
o Rapid processing time but poor quality.
 Enclonar, Kimberly / MLS 3A
o Methods of Freezing
▪ Liquid nitrogen
□ Most rapid
▪ Isopentane cooled by liquid nitrogen
□ Excellent for freezing muscle tissue.
▪ Carbon Dioxide gas
▪ Aerosol sprays
□ Does not freeze muscle tissues.
o Preparation of frozen sections
▪ Cold knife procedure
▪ Cryostat procedure
□ Simplest and quickest
o Applications:
▪ Rapid diagnosis during surgery
▪ Diagnostic and research enzyme
histochemistry
▪ Diagnostic and research demonstration of
soluble substances such as lipids and
carbohydrates
▪ Immunofluorescent and
immunohistochemistry staining
▪ Some specialized silver stains, particularly
in neuropathology

Special Processing Techniques

• Freeze Drying
o Rapid freezing (quenching) at -160C and
removing ice water molecules (dessication) by
transferring the still frozen tissue blood into a
vacuum chamber at a higher temperature
(sublimation)
• Freeze Substitution
o Process of dehydration
o Fixed in Rossman's formula or in 1% acetone and
dehydrated in absolute alcohol.

Gross Examination
Criteria for Rejection
• Discrepancies bet. the requisition of specimen labels
• No labels
• Leaking specimen containers
• No clinical data or history
• Inappropriately identified specimen

Materials
• Cutting tools: knife, blade
• Cassettes
• Filter paper
• Multi-colored ink set
• Forceps

Dimension: 3 x 2.5 x 0.4 CM

Procedure
1. Identify types of specimen
• Left breast
2. Structure included
• With tumor
3. Dimensions
• Length by width by height
• CM
4. Weight
5. Shape
6. Color
7. Consistency
8. Surgical margin, whether included or not involved
by tumor.