CYTOPA

THOLO

GY-8

DR

. M

AH

A A

L-SED

IK

Sputum , bronchial wash and bronchial

brush cytology

Objectives:1- introduction.

2- Safety in work.

3- Role of cytology in respiratory disease.

4- Cytological sample methods.

5- Sputum.

6- Hemoptysis

7- Importance of sputum culture.

8- Sputum collection and processing.

9- Bronchial washing.

10- Bronchial brushing.

Introduction: Cytological examination of specimens obtained from

the respiratory tract is a primary and frequently the

initial diagnostic technique performed in patients

with respiratory symptoms or in those presenting

with a pulmonary abnormality.

Due to the complexity of the respiratory tract and the

location of various target lesions, a variety of

cytological techniques have been developed for the

study of diseases involving the respiratory system.

Safety at Work:

Laboratory staff processing pulmonary cytological

materials are at particular risk of exposure to aerosol

infections.

They must protect themselves from this potential

hazard by wearing protective masks and gloves during

dealing with the samples, and following standard

laboratory hygienic procedures.

Major role:• Diagnosis of malignant neoplasms involving lung

both primary and metastatic.

Minor role:• Specific inflammatory process.

• Benign neoplasms.

Role of cytology in respiratory disease

Cytological Sampling Methods in respiratory

system:Sputum: from a spontaneous deep cough, obtained on

arising in the morning.

Bronchial Brushings: using bronchoscope.

Bronchial Washings: using bronchoscope.

Sputum

It is a mucous layer that covers the airways and

protects bronchial epithelium against inhaled noxious

substances.

Sputum is a mucous substance that is secreted into

the airways of the respiratory tract (lungs ,bronchi,

trachea) and can be coughed up, spit out or

swallowed.

It is produced by surface epithelial cells and sub

mucous glands.

Sputum Cytology

The spontaneous production of significant amounts of

sputum often indicates pulmonary disease.

Sputum is composed predominantly of mucoid

substances, and variable numbers of inflammatory

and epithelial cells.

Variations in the numbers of macrophages,

neutrophils, and epithelial cells and morphologic

alterations in the latter elements can yield significant

insight into the underlying pathologic process.

Chemical composition:

95% water

5 % organic components:

Squamous epithelial cells.

Respiratory epithelial cells.

Polymorphonuclear leukocytes.

Mononuclear cells.

Alveolar macrophages.

Carbohydrates ( such as sialic acid which contribute to

its high viscosity).

Proteins ( enzymes, immunoglobulins), glycoproteins.

Lipids.

5 CELLS3 FOOD

Abnormality of sputum:

Normal sputum is either white or colorless.

Yellow to green sputum can be an indication of pus,

infection such as pneumonia.

Blood in sputum is called hemoptysis which could be

due to e.g. ; lung cancer, tuberculosis, lung abscess ,

hemorrhage.

Parasites in sputum can occur as in ascaris.

Hemoptysis:

The expectoration (coughing up) of blood or of blood

stained sputum .

1 •TB

2 •Chronic bronchitis

3 •Cancer

4 •Pneumonia

5 •Trauma

6 •Autoimmune disease

Common causes of hemoptysis

Sputum

Culture

cytology

Importance of culture:

Detect and identify bacteria or fungi causing an

infection (such as pneumonia or tuberculosis).

Identify the best antibiotic to treat the infection

(sensitivity testing).

Monitor treatment of an infection.

Importance of Sputum Cytology:

Major role:• Diagnosis of malignant neoplasms involving lung

both primary and metastatic.

Minor role:• Specific inflammatory process.

• Benign neoplasms.

Bacterial cells in a sample of sputum

Collection Procedure:

a. Patient Preparation:

• The optimum time for specimen collection is within

15 to 30 minutes after waking and before eating

breakfast.

• Brushing of teeth or rinsing of the mouth with water

will reduce contamination by saliva.

• Instruct the patient to inhale and exhale deeply

forcing air from the lungs using the diaphragm.

Repeat until the patient coughs and is able to

produce a sputum specimen.

b. Collect the specimen in the sterile container,

attempting to obtain at least one teaspoon of sputum.

Saliva is of no diagnostic value.

Greater diagnostic yield may be obtained if specimens

are submitted on three to five successive mornings.

When sufficient amounts of spontaneously

produced sputum are unobtainable,

specimens may be induced by inhalation of a

nebulized solution composed of 15% sodium

chloride, with or without 20% propylene

glycol, or simply 3–8% sodium chloride

heated to 115°F.

c. Add 70% alcohol as soon as possible in a volume

equal to the specimen collected. Label each

container with the patient name, site, source and

requisition peel off number.

d. STORAGE: Submit the specimen at room

temperature.

The most popular techniques in handling sputum are

the ‘‘pick-and-smear’’ technique and the Saccomanno

methodology.

I. pick-and-smear:

Experience is essential to pick out significant areas

for processing.

They are prepared as direct smears for immediate

fixation (95% ethyl alcohol or spray fixation) then

stained with hematoxylin-eosin.

Saccomanno methodology:

When a significant delay is anticipated between

specimen taking and laboratory processing, prefixation

and processing by the Saccomanno method are

preferred.

Cells are collected in 50% ethanol and 2% polyethylene

glycol (carbowax).

Upon receipt in the laboratory, a blender is used to

emulsify the specimen, which is subsequently

centrifuged and prepared as smears.

Curschmann`s spirals are coiled strand of mucus. On Papanicolaou stains, they appear as purple helices. They are a non-specific finding associated with chronic respiratory disorders.  

Charcot-Leyden crystals are rhomboid shaped, orangiophilic structures deriving from degenerated eosinophils in patients with severe allergic disorders such as asthma.

Charcot-Leyden crystals

Strongyloides stercoralis

Candida

Granulomatous Inflammation (Tuberculosis)

Vegetable (plant) cells

Bronchial Washings and Brushings

Indications:

Bronchial brushings and washings are complementary to

sputum cytology in the diagnosis of pulmonary lesions.

The most common indications for bronchoscopy: Persistent cough.

Radiographic documentation of a new solitary

pulmonary nodule.

Un diagnosed hemoptysis.

Persistent localized wheezes.

persistent infiltrates on chest x-ray.

Bronchial washing

is part of a procedure called a bronchoscopy, in which a

physician looks into the lungs with a fiber optic

bronchoscope to check for irregularities and take tissue

samples.

Technique:

The physician injects saline through the bronchoscope

into the lung and then suctions it back out.

By checking the wash return fluid, the doctor can

diagnose bleeding, pneumonia, industrial pollutants,

fungal infections and different kinds of lung cancer.

Patients undergoing bronchial washing usually

receive topical anesthesia with sedation. 

Bronchial brushing:

is a procedure in which cells are taken from the inside

of the airway mucosa or bronchial lesions.

Flexible brushes are passed through the bronchoscope,

and the bronchial surface is gently abraded to obtain

the specimen.

A bronchial brushing is used to find cancer and changes

in cells that may lead to cancer. It is also used to obtain

specimens for microbiologic diagnosis.

Upon withdrawing the brush, agitate the brush

vigorously in a 5 to 10 mL vial of sterile saline or

fixative. DO NOT APPLY THE BRUSH DIRECTLY TO

SLIDES. If possible, detach the brush and leave it

in the vial and better to be examined in 4 hours.