Dan Dalan

Date of Award


Document Type


Degree Name

Master of Science (MS)




Five currently used flagella stains and staining procedures have been studied and evaluated. A staining method that is simplest, most reliable, and highly reproducible, especially for beginning students and technicians has been assembled and recommended.

Four important variables necessary for successful flagella staining were stressed. These variables are: growth of organisms, smear pre paration, staining procedure, and reagent stability.

Two important procedures which are part of some, but not all of the flagella staining methods studied are strongly recommended. One is the practice of enclosing a smear with wax pencil and adding a known, constant volume of dye. An effective ratio of staining surface area to volume of dye used is obtained with this procedure resulting in a stan dardization that greatly diminishes overstaining and understaining reactions. The second procedure is the use of a freshly mixed dye mordant staining mixture. This procedure is the key to reproducible production of good flagella stains that have clear backgrounds and few interfering artifacts. Dye and mordant reagents may be stored separate ly for long periods of time, probably indefinitely, and still provide good, clean, reproducible flagella stains when mixed just prior to use.