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AssigneeCenter National de la Recherche Scientifique (CNRS)
InventorsT. R. Strick, J. F. Allemand, D. Bensimon, A. Bensimon, V. Croquette
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An apparatus for manipulating and testing molecules comprising a surface to which the molecule is anchored at multiple points at one end, a paramagnetic bead on which said molecule is anchored at multiple points at its other end, a magnetic means for applying a force to the bead, said magnetic means being used to control the stretching and rotation of said bead and said molecule, an optical magnification means and a camera for the visualization of said bead, a computer means to which images from the camera are transmitted, said computer means comprising means for analyzing the motions of the bead, a rotating means for spinning the magnetic means about an optical axis of the magnification means, the magnetic means being positioned at a distance from the molecule in the direction of the optical axis of the magnification means, perpendicular to the optical axis, so that the force applied to the bead is substantially directed parallel to said optical axis, and the field produced by the magnetic means is perpendicular to the optical axis, wherein rotation and translation of the magnetic means causes rotation and translation of the bead, which allows to control stretching and torsion of the molecule.
1/ An apparatus for manipulating and testing molecules comprising
2/ The apparatus according to claim 1, wherein the computer means comprises
3/ The apparatus according to claim 2, wherein the computer means comprises means to verify that δx2=δy2.
4/ The apparatus according to claim 1 to 3, wherein the computer means comprises means to analyze a diffraction pattern of the bead to determine its fluctuations in the z direction of the optical axis of the magnification means.
5/ The apparatus according to claim 4, wherein the computer means comprise means to compare, in real time, a diffraction pattern of a bead to a reference set of diffraction patterns.
6/ The apparatus according to claim 4, wherein the reference set of diffraction patterns is previously built at 20 pN to 70 pN.
7/ The apparatus according to claim 4, wherein intensity profiles of diffraction patterns are obtained by averaging light levels over circles centered on the bead.
8/ The apparatus according to claim 1, wherein the surface is an internal surface of a capillary tube in which solutions are injected.
9/ The apparatus according to claim 1, wherein means to control the ambient temperature of the molecule and the bead is provided.
10/ A method for the manipulating and testing of molecules and in particular of DNA wherein
11/ The method according to claim 10, wherein
12/ The method according to claim 11, wherein the computer means verifies that δx2=δy2.
13/ The method according to one of claims 11 to 12, wherein a diffraction pattern of the bead is analyzed to determine its fluctuations in the z direction of the optical axis of the magnification means.
14/ The method according to claim 12, wherein the computer means compares, in real time, a diffraction pattern of the bead to a reference set of diffraction patterns.
15/ The method according to claim 14, wherein the reference set of diffraction patterns is previously built at 20 pN to 70 pN.
16/ The method according to claim 13, wherein intensity profiles of diffraction patterns are obtained by averaging light levels over circles centered on the bead.
17/ The method according to claim 10, wherein the surface is an internal surface of a capillary tube in which solutions are injected.
18/ The method according to claim 10, wherein the ambient temperature of the molecule and the bead is controlled.
19/ The method according to claim 10, wherein the molecule is covered with biotin at one of its ends and with digoxigenin at its other end, the surface being covered with antidigoxigenin, whereas the bead is covered with streptavidin.
20/ The method according to claim 10, wherein the elasticity of a molecule is changed by rotating the beads around the magnification axis.
21/ The method according to claim 11, wherein the elasticity of the molecules is first determined to select the bead(s) on which just one molecule is attached.
22/ The method according to claim 10, wherein the monitoring of the force or of the extension of the molecule is used to achieve sequencing.
23/ The method according to claim 10, wherein the activity of enzymes which interact with DNA by twisting or coiling can be measured in real-time.