Automated Field Fluorescence Microscope Using Compact Disc Technology for Single Biological Particle Detection

Systems & Processes Engineering Corporation (SPEC) and Dr. John Taboada of Taboada Research Instruments propose to design and provide proof-of-concept for an Automated Field Fluorescence Microscope (AFFM) instrument to rapidly and specifically identify individual biological particles in complex samples taken from the air, water, or blood. The proposed instrument combines the advantages of high-precision compact disc (CD) technology, fluorescence staining of nucleic acids, immunofluorescence and confocal microscopy. The proposed instrument will identify pathogenic bacteria or viruses by detecting the fluorescence from membrane permeable nucleic acid stains. The AFFM will use this fluorescence signal to focus on areas of interest. Using confocal microscopy, the AFFM creates a high resolution 3D image of the target location while simultaneously measuring the fluorescence from a second, spectrally discriminating dye-labeled antibody. This dye-labeled antibody, though naturally weaker in signal, will be highly specific to a biological pathogen. This process can take place in a matter of minutes and provide highly specific detection of targets. In Phase II, SPEC plans to design, construct and test a fieldable instrument which can rapidly and specifically identify individual biological particles, and which can be combined with an air or water sampler for remote. stand-alone monitoring of BW agents. BENEFITS: The AFFM instrument provides a capability to detect and identify single biological particles using a inexpensive compact disc based confocal microscope and fluorescent probes. The instrument developed in Phase II of this SBIR program has the potential to become a widely used commercial product with utility for field measurements and research in both the military and civilian sectors. Improved rapid methods for identifying pathogenic bacteria and viruses would be useful for a variety of applications.