Gram stain is a method of testing for microorganisms (bacteria) using a special series of stains. In this test, a sputum specimen is examined under the microscope. Sputum is the mucous secretions produced by the lower respiratory tract (in the lungs).

The gram stain method can be applied to almost any specimen, and is one of the most commonly used techniques for the rapid diagnosis of bacterial infections.

A sample of sputum is applied in a very thin layer to a microscope slide -- this is called a smear. A series of stains called a gram stain is applied to the specimen. It is first stained with crystal violet stain, then iodine, then decolorized, then stained with safranin.

The stained smear is then examined under the microscope for the presence of bacteria. The color, size, and morphologic appearance (shape) of the cells help identify the infecting organism.

You will produce a sputum sample from your lungs by coughing a specimen of mucus (not saliva or spit from the mouth) from deep inside your lungs into a container. If coughing does not produce sputum, a breathing treatment may precede the test to induce or encourage sputum production.

If you have a dry cough or are unable to produce a specimen, bronchoscopy may be necessary.

The person needing the test will usually have a productive cough (one where the person coughs up sputum). Occasionally, the sputum is collected by bronchoscopy, and more discomfort is associated with this procedure.

The test is performed when there is a persistent or prolonged cough, sputum has a foul odor or unusual color, respiratory disease is suspected, or there is a generalized infection.

No presence of organisms is normal. The sputum is clear, thin, and odorless.

A bacterial infection may be present. The infecting organisms may be tentatively identified. Culture is needed to confirm the diagnosis.

There are no risks unless a bronchoscopy is used to obtain the specimen.

The test may need to be repeated if the specimen produced contains only saliva from the mouth.