Lugol's Solution Stabilized with PVP for Gram Stain

Lugol's solution is an aqueous solution, which contains iodine (I2) and potassium iodide (KI) in a ratio of 1:2.

Used for Gram staining, Lugol's iodine solution is required to form the dye-iodine complex of Gram-positive bacteria. It can also be used as an antiseptic and disinfectant for emergency disinfection of drinking water, and as a reagent for starch detection in routine laboratory and other medical tests.

Stock solutions of I2 - KI in water are unstable. Iodine can be lost by evaporation and that can influence the result of the staining as well as affecting the working conditions in that area. Stainless steel parts of staining instruments can also be affected by the iodine in the Lugol's solution. If the concentration of iodine in the Lugol's solution decreases, the bacterial smears may become more susceptible to decolorization.

The problem of iodine loss can be overcome by using polyvinylpyrrolidine (PVP) in the I2 - KI solution. PVP forms a complex with iodine. This complex is more stable and has a longer shelf life. Material treated with Lugol's solution stabilized with PVP will give identical results with regard to differentiation of Gram-positive and Gram-negative bacteria compared to the standard Lugol's solution.

In the standard Gram procedure, the aniline dyes in the cell tissue of microorganisms form a red dye-iodine complex when exposed to iodine. In Gram-positive microorganisms, the dye-iodine complex cannot subsequently be dissolved from the cells by decolorizing agents such as alcohol or acetone. The cells remain dark blue in color. In Gram-negative microorganisms, the dye-iodine complex is dissolved and the cell turns red as a result of counterstaining.

Both Lugol's solutions (the standard type and the stabilized type) are available for bacteriological staining from Merck KGaA (Darmstadt, Germany), or EMD Chemicals (Gibbstown, NJ, USA) in the USA. Suitable protocols are available on the internet or on demand.

Related Links:
Merck KGaA
EMD Chemicals