LAB DIAGNOSIS OF

NEOPLASIA

Dr Tooba Khan
Lecturer
Dept. of Pathology
 Objectives:
• At the end of the lecture, the students will be able
to:
• List the various laboratory techniques that help
in the diagnosis of tumours
• Discuss the tumour markers used in the
diagnosis and management of cancers.
• Describe the role of molecular analysis in prompt
diagnosis of cancers.
 Techniques:
• Radiology
• Histology and cytology methods
• Immunohistochemistry
• Flow cytometry
• Circulating tumour cell
• Molecular diagnostics /Cytogenetics
• Molecular profiling
• Tumour markers
Radiology :
• X-Ray
• Fluoroscopy
• CT scan
• Ultrasound
• Magnetic resonance imaging (MRI)
• Nuclear medicine (gamma camera, Positron
emission tomography (PET)
• Interventional radiology
Histology & Cytological Methods:

• Cytology is the study of individual cells of

the body, as opposed to histology which is the
study of whole human tissue itself.
CYTOLOGICAL METHOD :
• Cytology tests may be used for diagnosis
or for screening:
1. Fine needle aspiration
2. Cytology tests on body fluids (Exfoliative
cytology)
3. Scrape or brush cytology (Abrasive
cytology)
4. Washings and lavage
 FINE NEEDLE ASPIRATION
CYTOLOGY (FNAC)
•There are two techniques for FNAC:
1. Direct FNAC.
2. Image guided FNAC by Ultrasound or CT scan.
•It is the popular method used for the diagnosis of palpable
tumors, e.g.
 Lymph nodal tumors
 Breast tumors
 Salivary gland tumors
 Thyroid tumors
 EXFOLIATIVE CYTOLOGY
• Tumour cells are less cohesive, hence are shed
from lining epithelium to body cavities, so they
can be collected and studied.
1. Cervical smear.
2. Urine containing cells from genitourinary tract.
3. Sputum.
4. Effusions in the body
 ABRASIVE CYTOLOGY
• Surface scraped using Ayre’s spatula or brush.
With or without optic guidance.

1. Cervical smears (PAP smear)

2. Bronchoscopic brushings
3. Endoscopic brushings from lesion of GIT
 WASHINGS AND LAVAGE
• Normal saline is instilled into body cavity and
aspirated back, collecting shed cells.

1. Gastric lavage.
2. Peritoneal lavage.
3. Bladder lavage
 MERITS / DEMERITS OF
CYTOLOGICAL DIAGNOSIS
ADVANTAGES DISADVANTAGE S

Rapid Small sample size and

sampling errors.

No hospitalization

Minimally invasive Cannot comment on

architecture and invasion.
 HISTOLOGICAL DIAGNOSIS
For histological diagnosis the following methods of
sampling is done:
 Biopsy- a surgical removal of small piece of tissue
for microscopic examination to detect the presence
of cancer cell.

There are three ways tissues can be removed:

1) Endoscopy.
2) Needle biopsy.
3) Surgical biopsy.
 FROZEN SECTION:
• Cryostat used in frozen sections is Liquid Nitrogen (-190 C) or
Liquid carbon dioxide at (-90 C) used to freeze IC water into
ice (embedding medium)
• Used for:
1. Rapid ‘on-table’ diagnosis.
2. Preservation of enzymes and labile substances like lipids
and glycogen.
3. Determining nature of mass lesion.
 HEMATOLOGICAL DIAGNOSIS
• Marrow is aspirated by bone marrow aspiration
needle.
• Biopsied by trephine biopsy needle.
• USED:
1. In the diagnosis of Leukemia
2. Metastasis from lymphoma or solid tumours.
This is needed for staging Leukemia.
IMMUNOHISTOCHEMISTRY:
• A method for detecting antigens or haptens in cells
of a tissue section by exploiting the principle of
antibodies binding specifically to antigens in
biological tissues.

• USED:
1. To help diagnose diseases
 ADVANTAGES:
1. Categorization of undifferentiated malignant tumor
2. Specific typing of leukemia /lymphoma.
3. Determination of site of origin of a metastatic tumor.
4. Detection of molecules that have prognostic &
therapeutic significance, e.g., ER-PR receptors in
carcinoma breast.
5. Expression of protein products of oncogenes.
6. Differentiating benign from malignant lesions
 FLOW CYTOMETRY:
• Flow Cytometry is a technique

• Used:
1. to detect and measure physical and chemical
characteristics of a population of cells or
particles.
2. Useful in the identification &
classification of lymphoma
and leukemia
 MOLECULAR DIAGNOSIS
• Detect molecular differences between diseased and
normal cells and detect cancer-specific alterations in
proteins, DNA, and RNA in body fluids
• useful in detection of minute translocations & minimal
residual disease.
 MOLECULAR TECHNIQUES
1. Polymerase chain reaction (PCR)
2. Fluorescence in situ hybridization (FISH)
3. DNA microarray analysis
4. Gene chip method
5. Tumor Profiling
 Molecular Profiling:
• “Tumour Genomic Profiling”
• Is a form of testing that classifies tumours based
on this genetic make-up to help diagnose and
treat cancer.
• Used:
1. As blood test or biopsy
2. Helps examine the DNA of cancer cells, looking
for genetic mutations that have been acquired
by these cells.
 TUMOUR MARKERS
• Some tumours release substance is called tumour
markers.
• Blood test can be performed to detect the blood
cells as well as for specific tumour markers.
• Tumour markers are biochemical indicators of
the tumour .
1. Antigens
2. Cytoplasmic proteins
3. Enzymes
4. Hormones
 ADVANTAGES:

• Screening and early detection of cancer

• Aid in the diagnosis of cancer
• Determine response to therapy
• Prognostic indicator of disease progression
• indicate relapse during follow-up period
 DISADVANTAGES :
• Lack of reliability
• Normal cells as well as cancer cells can produce
most tumor markers
• Proteins and/or modified proteins may vary among
individuals
• Tumor markers are not always present in early-
stage cancers
• Even when tumor marker levels are high, they are
not specific enough to confirm the presence of
cancer.