The sequence of events involved in the transition from attached liposomes to bilayer patches on hydrophilic and hydrophobic solid supports were visualized in situ by Tapping Mode atomic force microscopy in liquid. In a smooth manner, the attached liposomes spread and flattened from the outer edges toward the center until the two membrane bilayers were stacked on top of each other. The top bilayer then either rolls or slides over the bottom bilayer, and the adjacent edges join to form a larger membrane patch. This is clearly visible from the apparent height of 6.0-7.5 nm of the single bilayer, measured in situ. The addition of calcium appeared to increase the rate of the processes preventing the visualization of the intermediate stages. The same intermediate steps appeared to be present on hydrophobic surfaces, although the attached liposomes seemed to be distorted and the resultant membrane edges were uneven. This work has provided visual and detailed information on liposome coalescence (fusion) onto solid supports and demonstrated how the atomic force microscope can be used to study the process.
This paper presents the status of an ongoing development of a point detector for biological warfare agent sensing based on ultraviolet laser-induced fluorescence from single particles in air. The detector will measure the fluorescence spectra of single particles in a sheath flow air beam. The spectral detection part of the system consists of a grating and a photomultiplier tube array with 32 channels, which measure fluorescence spectra in the wavelength band from 300 nm to 650 nm. The detector is designed to measure laser induced fluorescence from single laser pulses and has been tested by measuring fluorescence from simulants of biological warfare agents in aqueous solution. The solutions were excited with laser pulses at the wavelengths of 293 nm and 337 nm. The paper also presents preliminary results on the sheath flow particle injector and time-resolved measurements of fluorescence from biological warfare agent simulants in solution.
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