Nanoscale motion detector
a motion detector and nano-scale technology, applied in the field of nano-scale motion detectors, can solve the problem of limited use of state-of-the-art sensors
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example 1
Drug Affinity Detection
[0040]These experiments involve Topoisomerase II (TopoII—FIG. 3) and its interaction with anticancerous drugs. TopoII is an essential enzyme that interacts with DNA to simplify its topology and permits the transcription to occur safely.
[0041]This enzyme requires ATP to modify its 3D conformation and to act on DNA. TopoII is also the preferred target of numerous anticancerous drugs such as aclarubicin. This drug binds to TopoII, freezes its conformation and inhibits its action (15). In the first experiment, TopoII was adsorbed onto both sides of a cantilever. It was than introduced in the analysis chamber of an AFM and its laser beam was collimated on the apex of the cantilever. The reflection of the laser beam, sent to a split photo-detector, allowed detecting the fluctuations of the cantilever as depicted in FIG. 1 and, more in detail, FIG. 3.
[0042]The experiment consisted in injecting successively an ATP depleted buffer, an ATP enriched solution and an aclar...
example 2
Detection of Biochemical Reactions
[0048]As mentioned previously, TopoII interacts with DNA to simplify its topology. To record this reaction with our method we deposited TopoII-supercoiled DNA complexes on both sides of an AFM cantilever, as depicted in FIG. 6.
[0049]The experiment was performed using an APTES-coated AFM cantilever. The different media injected during experiment are: buffer (50 mM Tris-HCl (pH 8.0), 150 mM NaCl, 10 mM MgCl2 and 0.5 mM dithiothreitol), 0.02 mM AMPPNP (an ATP analogue) and 0.02 mM ATP. Panel A shows the cantilever deflection data. Panel B depicts the experimental set-up to follow TopoII-DNA interactions. a) DNA molecule, b) TopoII, c) AFM cantilever, d) laser beam. Panel C evidences the differences in terms of the variance.
[0050]Here again the cantilever covered with TopoII-supercoiled DNA complexes was inserted in the analysis chamber and exposed to several consecutive liquid environments: 1) ATP free solution, 2) buffer containing a non functional AT...
example 3
Antibiotic Sensitivity Detection
[0051]The presented method is sensitive enough to detect external as well as internal motion of bacteria and can be used to assess the action of antibacterial agents. This experiment shows the capability to explore the sensibility of bacteria to antibiotics with a very high temporal resolution ranging between seconds and minutes.
[0052]Motile bacteria (Escherichia coli), resistant to kanamycin but sensitive to ampicillin, were adsorbed to both sides of a cantilever. The bacteria were successively exposed to both antibiotics to determine the impact on the bacterial motion / viability (see FIG. 7). In these experiments the cantilever was introduced in the analysis chamber and exposed to a solution containing bacteria that eventually attached to its surface. The system was then exposed to: 1) nutriment depleted buffer, 2) nutriment solution (Lysogeny broth (LB)), 3) kanamycin-enriched LB solution 4) LB solution 5) ampicillin-enriched LB solution and, finall...
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Abstract
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