Sputum

examination

Prepared by: Eliazel G. Ocfemia

SPUTUM
SPUTUM
Secretion produce in the lungs and bronchi
Produce when a person’s lungs are
diseased or damage
Sometimes called phlegm
Coughed up from the lungs
Clinical
significance
Detection of :
Occurrence of infections (TB, URTI)
Respiratory diseases (asthma, Inflammation)
Neoplastic diseases of lungs, other respiratory
organs
Specimen collection
Patient preparation:

Collect sputum Prepare clear and

before antibiotics sterile disposable
are given container
Rinsed mouth
with distilled
water
Specimen
collection

Expectoration or sputum
production
First morning- ideal
Specimen Collection
Specimen
collection
In children – 3methods
Nasopharyngeal swab,
Cough plate,
Cough swab

Tracheal aspiration- deliberated patient

Gastric washing- absence of sputum
Normal
Sputum pH:

6.5- 7.0
Color:
• Colorless and translucent
• Whitish to faint yellow
• Orange to purulent
Odor

Odorless
Macroscopic examination

Amount :
Scanty: bronchial asthma, acute
bronchitis, early pneumonia
Ample: bronchiectasis, lung abscess,
edema
Color:
White/gray (may be normal)
color changes associated with other conditions

Dark yellow/ green

bacterial infection of the lower respiratory tract
Green sputum- pneumonia
Yellow green- cystic fibrosis
Macroscopic examination
Color:
Olive green/grass green
Carcinoma of the lungs

Bright green
Pseudomonas infection Jaundice,
Rupture of liver
Caseous pneumonia

Macroscopic examination
Color:
Brown (presence of tar)
Found in smokers
Brown- black lung disease ( pneumoconiosis)
Brown/black- presence of blood

Macroscopic examination
Color:
Pink
Frothy pink- pulmonary edema
Blood-tinged- tuberculosis

Bloody
Pulmonary embolism

Macroscopic examination
Color:
Anchovy/ rusty
Early lobar pneumonia
Tuberculosis
Pulmonary gangrene
Hemorrhage (pulmonary infarction)

Macroscopic examination
Color:
Prune juice
Late pneumonia

Whitish yellow
Indicates pus

Macroscopic examination
Consistency
Mucoid
Viral infection

Purulent
Chronic bronchitis / bacterial infection
Serous or Frothy
Lung edema
Macroscopic examination
 Consistency
Putrid
• Lung abscess,
• Bronchiectasis,
• Gangrene of the lung

Macroscopic examination
 Consistency
Sweetish
• Pulmonary tuberculosis with
cavities,
• Broncho moniliasis,
• Bronchiectasis

Macroscopic examination
 Consistency

Cheesy
• Necrosis of malignant tumor
perforating emphysema

Macroscopic examination
 Microscopic Examination
Transfer specimen to clear slides.
Smear made on clear slides should be
air dried, fixed over flame and stained.
Examine under the microscope

Wet preparation – Entamoeba, Paragonimus

Wet staining wt methylene blue – malignant cells
 SPUTUM
SLIDE
Different stains used:
Gram’s stain/ ziehl – Neelsen stain= For AFB
Wright’s stain= for blood cells
Buffered crystal violet= for epithelial cells
Pap’s stain= cytology of sputum

Microscopic Examination
 Gram Stain
Stain procedure:
 Crystal violet – primary stain
1min/rinse
 Gram’s iodine – mordant
1 min/rinse
 Alcohol – decolorizer
5 seconds
 Safranin – counterstain
1 min, rinse Microscopic Examination
 Types of Cells
Neutrophils: pyogenic infection
of respiratory tract

Eosinophils: asthma and

parasitic infections of lung

Microscopic Examination
 Types of Cells

Lymphocytes: early pulmonary

tuberculosis

RBC’s: hemorrhage into the lungs

Microscopic Examination
 Types of Cells

Anthracotic pigment laden cells: coal

workers, pneumoconiosis, smoky
polluted areas

Microscopic Examination
 Types of Cells
Heart failure cells : large
mononuclear cells with brown blood
pigment

Microscopic Examination
 Others

Elastic fibers
From the wall of alveoli and
bronchiol
TB, lung abscess, lung gangrene

Microscopic Examination
 Others
Crystals: Charcot - Leyden crystals
Needle like, colorless
Most significant and characteristic
of bronchial asthma

Microscopic Examination
 Others

 Myelin globules
Fat droplets with no significant

 Yeast : not significant

Microscopic Examination
 Others

Pathogenic fungi
Candida albicans, Coccidioides
immitis, sulfur granules
Cryptococcus neoformans,
geotrichum (normal)

Microscopic Examination
 Others
Bacteria, parasites, viruses
Need special virus laboratory method

Neoplastic cells
Stained by papanicolaou technique
and examine

Microscopic Examination
 Pap’s staining Procedure
Distilled Water 5 dips
Distilled Water 10 seconds
(2 changes)
90% Alcohol
(Fixation)15-30
minutes
3 Distilled Water
10 seconds
7
1 5

2 4 6
60% Alcohol Hematoxylin stain 1% Acid Alcohol 10 seconds
2 minutes 3 minutes (1 dip)
Microscopic Examination
 Pap’s staining Procedure
60% Alcohol 95% Alcohol
2 minutes 2 minutes
Scott’s Tap Water 95% Alcohol
2-3 minutes
10 2 minutes 14
8 12

9 11 13
Running Tap Water 80% Alcohol 95% Alcohol
2 minutes 2 minutes 2 minutes
Microscopic Examination
 Pap’s staining Procedure
Absolute Alcohol Mount in D.P.X
2 minutes (3 changes)
Eosin Azure Stain
3 minutes 1 Xylene 2 minutes
(2 changes) 2
1 7 1 1
5 9
1 1 2
6
95% Alcohol
2 minutes (4 changes)
8
Absolute
Alcohol+Xylene (1:1) 2
Xylene0
Till clear

minutes Microscopic Examination

Stained Cells’ Color
Nuclei : Blue
Acidophilic cells : Red
Basophilic cells : Blue Green
Erythrocytes : Orange-red
Keratin : Orange-red
 Stained Cells Color
Superficial cells : Pink
Intermediate and
Parabasal Cells : Blue Green
Eosinophil : Orange-red
Candida : Red
Trichomonas : Grey green
Miscelleneous Findings
Cheesy masses
Pulmonary gangrene/ tuberculosis
Bronchial cast
Branching tree like cast of bronchi
Composed of fibrin,
white or gray color
 Broncholiths
(lung stone)
Calcification of necrotic or
infected tissue
Chronic tuberculosis

Miscelleneous Findings
Dietrich’s plugs

Frequently observed in
putrid bronchitis and bronchiectasis
Composed of:
Cellular debris
Fatty acids
Crystals
Fat globules
Bacteria
Miscelleneous Findings
Cursch – man’ spiral

Bronchial asthma

Allergic
bronchopulmonary
aspergillosis

Miscelleneous Findings
 Sputum
examination

Prepared by: Eliazel G. Ocfemia