Polymerized LB films imaged with a combined atomic force microscope-fluorescence microscope

The first results obtained with a new stand-alone atomic force microscope (AFM) integrated with a standard Zeiss optical fluorescence microscope are presented. The optical microscope allows location and selection of objects to be imaged with the high-resolution AFM. Furthermore, the combined microscope enables a direct comparison between features observed in the fluorescence microscope and those observed in the images obtained with the AFM, in air or under liquid. The cracks in polymerized Langmuir-Blodgett films of lO,l2-pentacosadiynoic acid as observed in the fluorescence microscope run parallel to one of the lattice directions of the crystal as revealed by molecular resolution images obtained with the AFM. The orientation of these cracks also coincides with the polarization direction of the fluorescent light, indicating that the cracks run along the polymer backbone. Ripple-like corrugations on a submicrometer scale have been observed, which may be due to mechanical stress created during the polymerization process

Orange passion:Orangism in the political culture of the Netherlands (1780-1813)

In this book I have examined Orangism in the Netherlands in the years between roughly 1780 and 1813. By focusing on the political ideas of Orangists in this period, I exposed he complexity of Orangism. Orangist political discourse drifted in several directions. This was partly due to successive political changes that determined Dutch politics between 1780 and 1813. Orangists reacted differently to each event and development. The rise of the Patriot movement, the proclamation of the Batavian Republic and the increasing influence of the French emperor Napoleon Bonaparte in the Netherlands had a major influence on the lives and thinking of Orangists. Despite the radical changes in Dutch politics in these years, Orangism never disappeared. My research has shown that in all their diversity in every turbulent period, Orangists developed their own political ideas by adapting them to the circumstances of the time. In doing so, they helped shape the political culture of the Netherlands between 1780 and November 1813 and contributed to the foundation of the political system after 1813: that of a constitutional monarchy

Tip-sample interactions in atomic force microscopy: I. Modulating adhesion between silicon nitride and glass

An adhesive interaction between a silicon nitride AFM tip and glass substrate in water is described. This adhesion is in the range 5-40 nN, of which a large component is likely to be due to hydrogen bonding between the silanol groups on both surfaces. The interaction can be modulated by a variety of buffers commonly used in biochemical and biological research, including sodium phosphate, tris(hydroxymethyl)aminomethane, glycine, and N-2-hydroxyethyl-piperazine N'-2-ethanesulfonic acid. Using these buffers it appears that there are effects of ion concentration, ion type and pH on the measured adhesion. Of the conditions examined, phosphate was most effective at reducing adhesion and could be used at concentrations as low as 10 mM at neutral pH. The results demonstrate that the chemical interactions between tip and sample can be modulated, and provide a basis for designing conditions for imaging and manipulating biological molecules and structures

Natural dairy cow health

guide to keeping your herd healthy with herbs and other natural product

Atomic Force Microscopy of DNA Electrophoresed onto Silylated Mica

A new technique has been developed for electrophoresing DNA molecules from an agarose gel onto a silylated mica substrate where they can be imaged with an atomic force microscope (AFM). With a simple modification, the technique can also be used for polyacrylamide gels. This method does not require purification of samples from the gels. Using tapping mode AFM, we have observed plasmids after electrophoretic separation into two bands. Differences in conformation were observed between the plasmids in the two bands

Imaging Single-Stranded DNA, Antigen-Antibody Reaction and Polymerized Langmuir-Blodgett Films with an Atomic Force Microscope

The combination of an (AFM) atomic force microscope together with microfabricated cantilevers that have integrated tips opens many possibilities for imaging systems of great importance in biology. We have imaged single-stranded 25mer DNA that was adsorbed on treated mica or that was covalently bound with a crosslinker to a polymerized Langmuir-Blodgett (LB) film, the top monolayer of a bilayer system. At low magnification the AFM shows cracks between solid domains, like in an image taken with a fluorescence microscope. At higher magnification, however, the AFM reveals much finer cracks and at still higher magnification it reveals rows of individual molecules in the polymerized LB film with a spacing of 0.45 nm. We have also imaged a LB film consisting of lipids in which 4% of the lipids had hapten molecules chemically bound to the lipid headgroups. Specific antibodies can then bind to these hapten molecules and be imaged with the AFM. This points to the possibility of using the AFM to monitor selective antibody binding

Improved Visualization of DNA in Aqueous Buffer with the Atomic Force Microscope

An improved method has been developed for imaging deoxyribonucleic acid (DNA) in aqueous buffer with the atomic force microscope (AFM). DNA on untreated mica can be imaged in aqueous buffer with the AFM if the DNA is deposited onto the mica in a buffer with HEPES and MgCl2, if the sample is rinsed thoroughly with high water pressure, and if the imaging is done with an electron beam-deposited (EBD) tip that has been deposited in the scanning electron microscope (SEM). The water rinse removes DNA that is otherwise easily scraped off the substrate. There is evidence that sharper tips may be more damaging to DNA when imaged in aqueous buffer especially when the DNA is bound tightly to the mica. The ability to image DNA in nearly biological conditions has potential applications for imaging biomolecular processes with the AFM