Methods for detection of Leishmania in bone marrow smear

The reagents for the detection of Leishmania

• Methyl alcohol, or absolute ethyl alcohol, or a mixture of Nikiforov (mixture of equal amounts of ether and absolute ethanol).
• Phosphate buffer pH 6.9–7.1, for the preparation of which is mixed 40 ml KH₂RO₄ (9,078 g dry matter KH₂RO₄ in 1 liter of solution) and 60 ml Na₂PHO₄ (11,876 g dry matter 1 liter of solution).
• Azureozin Romanovsky.

Methods of detection of Leishmania in bone marrow smear

From bone marrow punctate immediately after taking care, not to crush or deform cells,preparing thin smears. If dense punctate, make imprint. When significant blood in punctate from it isolated on the glass a little whitish lumps of fat, if they are distinguishable, and prepare them smears.

The prepared smear is air-dried, then fixed, as the blood smears. Dry fixed smears can be stored for several days.

In smear poured diluted in buffer coloring Romanovsky (about 3-5 ml) and left for 30 50 m. After staining rinsed with distilled water, dried and examined under a microscope with immersion system.

Evaluation of the results of detection of Leishmania in bone marrow smear

In the midst of Leishmania disease usually found in the bone marrow punctate in a significant amount and, therefore, they are easy to find. However, at the early stage of the disease and in patients treated for, to identify the causative agent, It requires a long view of all smear (no less 40 m).

Leishmania found in macrophages or extracellularly as rounded or risovidnyh cells 3-5 microns in diameter. The cytoplasm is colored gray-blue color, and the core - in a reddish-purple. In the cytoplasm, located kinetoplasts - rod-shaped or rounded education, intensely colored core.

For Leishmania sometimes take platelets or "shards" of cells (Komocki citoplazmy, containing nuclear material), found in bone marrow smears, foreign microorganisms (eg, unicellular algae), trapped in a smear during painting. The stained smear are harder, than Leishmania (dark blue cytoplasm and nucleus of bright crimson), kinetoplasts missing.