HISTOPATHOLOGIC

TECHNIQUES

histotechniques
 definition
• Are laboratory procedures, processes,
methods, steps and/or techniques dealing
with tissue specimens to aid
pathologists in their diagnosis for the
presence or absence of malignancies in
tissues and organs of the body.
 Personnel composition of histopath
section / pathology dept:
• 1. Pathologists
• Are doctors of medicine who specializes in
pathology.
• Are licensed physicians who have
undergone special training in laboratory
medicine.
 Classification of pathologists:
1. Anatomical / surgical pathologist
• One who is trained in the gross
structures of man.
• Example:
• autopsy (dead body)
• Biopsy (living body)
 Classification…
• 2. Clinical pathologists
• One who ….
• specializes in clinical laboratory
medicine.
• is knowledgeable in the clinical laboratory.
• Heads a clinical laboratory.
classification….
• 3. Forensic pathologists
• Specializes in forensic medicine
• involved in medico-legal cases.
• One who determine causes of death due to
normal and abnormal circumstances.
• Also referred to as medicolegal
practitioners
Roles / functions of pathologists:
1. Practical application of knowledge
• Practical application of training undergone.
• Example:
• Microscopic examinations of tissue specimens
for diagnosis.
2. Sharing of knowledge to his colleagues and
other members of the medical and
paramedical team.
• CPC (Clinical Pathology Conference)
• 3. Supervision of a Clinical Laboratory
 Personnel Composition….
• 1. Pathologist/s
• 2. Medical Laboratory Scientists / Medical
Technologists
• Specific role of an RMT in histopathology section
of a Clinical Laboratory is to assist the pathologists
in the diagnosis based on the technical aspect only,
that is, proper processing of tissue specimens.
• 3. Laboratory Technicians
• Not a licensed MLS / MT
• Work under the supervision of an RMT and
Pathologist.
• 4. Laboratory aide
• Work under the supervision of a lab technician and
RMT.
Reminders to Path personnel!
• Each patient is unique so each
specimen is unique.
• Pathology personnel must be accurate,
reliable, well-organized, logical and
practical. Have an interest in medicine /
science and be able to recognize and
embrace the responsibility for the well-
being of others.
• http://www.ntpathology.com/careers_medic
al_science.html
 Basic equipments
1. Microscope
 Microscope
• Greatly magnifies very minute objects
• Study of cell morphology
• Important in the diagnosis of both
normal and pathological conditions of
cells and tissues.
 Types of Microscopes
1. Fluorescence Microscope
•
Fluorescence Microscope
 Ultraviolet Microscope
• Makes use of ultraviolet light through quartz
or reflecting lenses
Electron Microscope
Scanning electron microscope
Transmission electron
microscope
2. Microtome
Cold Microtome
Other equipments….
Float-out-bath
Paraffin oven
Incubator
Warming plate
Automatic honing machine
Automatic Tissue Processor
Vacuum Tissue Processor
Embedding Machine
Other materials
Other materials
Other materials
Other materials
 Reagents
• Fixatives / Fixing • Mounting agents
agents • Ringing agents
• Decalcifying agents • Honing agents
• Dehydrating agents • Staining solutions
• Clearing agents • Adhesives
• Infiltrating media
• Embedding media
 Tissue Specimens
• SOURCE
1.Surgical specimens
• *major operations
• *minor surgery : biopsy
• *Autopsy
2.Pap’s specimens
 Tissue specimens
• TYPES:
• 1. based on consistency / appearance
• Hard tissue – bone, calcified tissue
• Soft tissue – brain, lungs, kidney, eyes…
• Liquid – blood, lymph
• Fragmented – Pap’s specimen
 Types of tissue specimens
• 2. Based on condition or state of
specimen
• Fresh / viable
• Fixed / preserved
 Tissue Examinations
• May vary according to the following
factors:
1.Structural and chemical components of
the cells to be studied
2.The nature and amount of the tissue to
be evaluated
3.The need for an immediate examination
of a tissue structure
 Tissue Examinations
• Gross Structural / Microanatomical
Examination
• A. FRESH / VIABLE / LIVING TISSUES
• Are usually examined if there is an
immediate need for tissue evaluation
 Fresh…
• 1. Direct Observation
• a) Exteriorization
• b) Transillumination
• 3) Transparent chamber method
Transillumination
 Fresh….
• 2. Cell, Tissue and Organ Culture
 Fresh…
• 3. Mechanical Manipulation Methods
a)Micro-surgery
• Micro-surgical microscope
• Microprobe
• Microneedle
• Microhooks
• Micromanipulation to achieve controlled
movements in various planes
• Also referred to as: Tease Method or
• Dissociation
Method
1. microsurgery
2. Squash Preparation
 3. Smear preparation

a)Streaking
b)Cotton Swab Application
c)Spreading method
d)Pull-apart Method
e)Touch Preparation / Impression Smear
Smear prep…..streaking
Smear prep…..cotton swab
Smear prep….spreading
Smear prep….pull-apart
Smear prep….touch
preparation
4. Motion picture
5. Radiation probes
6. Differential Centrifugation
Differential Centrifugation
Differential Method
Differential method
 6.a. Zonal Velocity
Centrifugation
• Also referred to as Density Gradient
• Centrifugation
 7. Freeze-etched Method
• Freeze Fracture Method / Cryofracture
Method
Freeze-etch
Freeze-etch
Freeze-etch
 Next topic….
• Methods of tissue examination using fixed /
• preserved tissues……OK?