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The aim of the present work was to evaluate the topography of spirochetes’ cells Borrelia burgdorferi s.s. B31 in atomic force microscope (AFM). Results: The length of spirochetes B. burgdorferi has ranged between 15.38-22.68μm. The cells of spirochetes do not constitute structures of a fixed diameter and height. Thus, in order to identify real parameters of cells, the horizontal distance and vertical distance have been used in the measurements. The average value of a spirochetes’ diameter has been estimated by taking series of measures and it is 0.40 μm. The average value of a spirochetes’ height has been estimated by taking series of measures and it is 70.14 nm. The analysis of a relation between measured parameters of spirochetes: diameter and height revealed that along with the growth of diameter of a bacteria cell, its height also grows. The average value of a fibers’ diameter has been estimated by taking series of measurements and it is 0.09 μm and the average height of fibers was 7.91 nm. Conclusions: The atomic force microscope (AFM) is a modern tool with a broad spectrum of observatory and measure abilities and is a technique which has been used in biology and microbiology to investigate the topography of surface and in the evaluation properties of cells.
NC-AFM (non contact atomic force microscopy) was used to observe the surface of triticale starch granules. The method allows to obtain micro-images of high resolution with no sample pretreatment. Surface structures and protrusions or pores found on the surface of triticale starch granules had a broad range of diameters. The existence of flat, smooth regions on the starch granules, with no visible structures, pores or protrusions, was confirmed.
A new method based on combined atomic force microscopy (AFM) and fluorescence microscopy observations, is proposed to visualize the insertion of glycosylphos- phatidyl inositol (GPI) anchored alkaline phosphatase from buffer solutions into sup­ported phospholipid bilayers. The technique involves the use of 27 nm diameter fluo­rescent latex beads covalently coupled to the amine groups of proteins. Fluorescence microscopy allows the estimation of the relative protein coverage into the membrane and also introduces a height amplification for the detection of protein/bead com­plexes with the AFM. The coupling of the beads with the amine groups is not specific; this new and simple approach opens up new ways to investigate proteins into sup­ported membrane systems.
Using atomic force microscopy (AFM) we performed dynamic force measurements of the adhesive forces in two model systems: avidin-biotin and streptavidin-biotin. In our experiments we used glutaraldehyde for immobilization of (strept)avidin on the tip and biotin on the sample surface. Such interface layers are more rigid than those usually reported in the literature for AFM studies, when (strept)avidin is coupled with biotinylated bovine albumin and biotin with agarose polymers. We determined the dependence of the rupture forces of avidin-biotin and streptavidin-biotin bonds in the range 300-9600 pN/s. The slope of a semilogarithmic plot of this relation changes at about 1700 pN/s. The existence of two different regimes indicates the presence of two activation barriers of these complexes during the dissociation process. The dissociation rates and activation energy barriers, calculated from the Bell model, for the avidin-biotin and streptavidin-biotin interactions are similar to each other for loading rates > 1700 pN/s but they are different from each other for loading rates < 1700 pN/s. In the latter case, the dissociation rates show a higher stability of the avidin-biotin complex than the streptavidin-biotin complex due to a larger outer activation barrier of 0.8 kBT. The bond-rupture force is about 20 pN higher for the avidin-biotin pair than for the streptavidin-biotin pair for loading rates < 1700 pN/s. These two experimental observations are in agreement with the known structural differences between the biotin binding pocket of avidin and of streptavidin.
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