Quarterly Report for Contract N68171-94-C-9101 (Norwegian Radium Hospital).
Abstract
The Gram staining procedure is well established for separation of large groups of bacteria, i.e. into Gram positive and Gram negative ones. This procedure was developed for microscopic purposes and is thus applied on bacteria attached to slides. Other staining procedures for separation of large groups of bacteria may be based upon binding of various dye-labeled compounds to specific parts of the bacterial wall. Thus, an alternate Gram staining procedure is based upon a fluorescence4abeled lectin which binds specifically to N-acetylglucosamine in the outer peptidoglycan layer of Gram positive bacteria (Sizemore R.K. et al. Applied and Environmental Microbiology, 1990). The peptidoglycan layer of Gram negative bacteria is covered by a membrane which is not labeled by this lectin. This can be used to distinguish between Gram positive and Gram negative bacteria. In this alternate Gram staining procedure smears are made on glass slides, air dried and heat fixed, whereafter smears are covered with freshly thawed lectin solution for 30 5 and then gently rinsed with phosphate buffer. Cover slips are then placed on the wet slides. When analyzed in a fluorescence microscope (488 nm), Gram positive bacteria fluoresce bright yellow-green, whereas Gram negative bacteria do not fluoresce.
Document Details
- Document Type
- Technical Report
- Publication Date
- Jan 01, 1990
- Accession Number
- ADA297948