01854naa a2200169 a 450000100080000000500110000800800410001910000200006024500650008026000090014552013780015465000140153270000190154670000170156570000130158277300890159516212352001-07-26       2000    bl ---       0--  u    #d1 aGROSSART, H. P.  aA simple, rapid method for demonstrating bacterial flagella.  c2000  aWe developed a simple, rapid method for demonstrating flagellation of bacteria using the fluorescent protein stain NanoOrange (Molecular Probes, Eugene, Oreg.) The NanoOrange reagent binds to hydrophobic regions of proteins, which results in substancial enhacement of fluorescence. Unbound  reagent is essentially non-fluorescent. NanoOrange fluorescently stained bacterial cell bodies, as well as flagella and other appendages, which could be directly observed by epifluorescence microscopy. Detection of flagella was further improved by using a charge-coupled device camera for image capture and processing. The reliability of the method was tested by using 37 pure cultures of marine bacteria. Detection of flagella on the isolates by NanoOrange staining was compared to detection by transmission electron microscopy (tem). For 36 of 37 cultures, the two methods yielded the same results. In one case, flagella were detected by TEM but not by NanoOrange, although the difference may be attributable to differences between the culture preparations. NanoOrange staining is rapid (10 to 15 min) and does not require fixation or dehydration, solive samples can be stained. Since NanoOrange is a general protein stain and works directly in seawater, it may also prove to be useful for staining other proteinaceous material that is of interest to aquatic microbial ecologists.  aBactéria1 aSTEWARD, G. F.1 aMARTINEZ, J.1 aAZAM, F.  tApplied and Environmental Microbiology, Washingtongv. 66, n. 8, p. 3632-3636, 2000.