JP6973766B2 - Exercise measuring device - Google Patents

Description

The present invention relates to a motion measuring device that detects the motion of a target via a marker attached to the target, and in particular, detects a response from a label irradiated with a quantum beam through a detection window and evaluates the motion state of the label. The present invention relates to a motion measuring device capable of obtaining an index value.

A method called Diffracted X-ray Tracking (DXT), which measures molecular motion from the motion of a diffraction point, is known as a method for detecting the motion of a subject via a marker attached to the subject. (Non-Patent Document 1). In this X-ray single molecule tracking method, a label is attached by irradiating a marker attached to a biomolecule or other object with white X-rays and tracking the movement of the diffraction point of the X-ray diffracted by the label with an image sensor. It is possible to measure structural changes and other movements of the subject.

However, the white X-rays used in the X-ray single molecule tracking method are expensive and the equipment becomes large. Further, by using white X-rays, the irradiation flux becomes relatively large, and the damage to biomolecules and other objects becomes so large that it cannot be ignored.

"Diffracted X-ray tracking: new system for single molecular detection with X-rays" YC Sasaki, Y. Okumura, S. Adachi, Y. Suzuki, and N. Yagi, Nucl. Instrum. Methods Phys. Res. A 467, 1049 (2001).

The present invention has been made in view of the above background technology, and is inexpensive, easy to miniaturize, and moves the target while using a simple X-ray source or other quantum radiation source that can reduce damage to the target. It is an object of the present invention to provide a motion measuring device capable of reliably evaluating such things.

The motion measuring device for achieving the above object is a beam irradiation unit that irradiates a quantum beam on a marker attached to a target, a sensor that detects a quantum beam from the label, and a predetermined detection window set in the sensor. A signal extraction unit that extracts a signal, and a signal processing unit that determines an index value based on the change over time in the detection intensity of the signal by a predetermined detection window and evaluates the motion state of the sign based on the determined index value. To prepare for.

In the motion measuring device, if the wavelength range of the quantum beam is wide, the diffraction point moving across the detection window illuminates the predetermined detection window for a moment, and the signal corresponding to the diffraction point by the predetermined detection window is used. Since the index value is determined based on the change in the detection intensity of Measurement can be performed using a device with a small irradiation flux. Here, since the index value determined based on the change over time of the detection intensity by the detection window can be said to indirectly represent the movement of the diffraction point and the movement speed of the label, such an index value. The movement state of the marker can be appropriately evaluated based on the above. The quantum beam includes X-rays, electron beams, neutron beams and the like.

In a specific aspect of the present invention, in the motion measuring device, the sensor detects the quantum beam from the label as an image. In this case, the sensor will be an existing image sensor or an application thereof, and the measurement of the quantum beam from the label will be high-speed and highly reliable.

In another aspect of the invention, a predetermined detection window is pixels arranged in a matrix on the detection surface of the sensor, in which a detection area defined by one pixel or a plurality of pixels on the detection surface is set in the sensor. Will be done. In this case, the signal is detected in the detection area in which one or more detection windows for each pixel are combined. By using the detection window as a pixel, signal processing becomes simple. Further, when a plurality of detection windows having a plurality of pixels are set as a detection area, information from a plurality of pixels having substantially the same structure and state can be uniformly processed, so that signals from the plurality of detection windows can be uniformly processed. Alternatively, the process of integrating information and the process of extracting statistical information become relatively easy.

In yet another aspect of the invention, the detection area is set to an arrangement and size according to the characteristics of the label. By setting the detection area in consideration of the diffraction conditions of the label attached to the target, efficient measurement with the target narrowed down becomes possible.

In yet another aspect of the invention, the index value relates to variations in detection intensity over time. If the speed of the diffraction point is sufficiently slow with respect to the detection time per detection window, it can be said that the variation in the change over time represents the length of stay of the diffraction point. Indirectly represents the moving speed of the diffraction point and thus the moving speed of the label.

In yet another aspect of the invention, the index value is the standard deviation. Measurements using standard deviation are statistically reliable.

In yet another aspect of the invention, the index value is the rate of change in detection intensity. When the speed of the diffraction point is relatively fast with respect to the detection time per detection window, the rate of change in the detection intensity indirectly represents the movement speed of the diffraction point or the motion speed of the label. .. As the rate of change in the detected intensity, for example, a time derivative value of the amount of change in the detected intensity can be used.

In yet another aspect of the invention, the beam irradiator irradiates monochromatic X-rays as a quantum beam and the sensor detects the X-rays diffracted by the label. In this case, the label will have X-ray diffraction characteristics. In addition, the diffraction image from the label can be measured with a relatively simple facility.

In yet another aspect of the invention, the label is a nanocrystal. In this case, a relatively high diffraction signal can be obtained while making the label compact and lightweight. In particular, nanocrystals show interlocking that reflects their motion by adhering to macromolecules and other objects in place, so that the motion of macromolecules and the like can be accurately grasped.

In yet another aspect of the invention, the rotational speed of the label is evaluated as the motion state of the label. When observing a diffraction image, the rotation speed of the label is basically evaluated.

In yet another aspect of the present invention, the signal processing unit has a conversion unit that converts an index value into a rotation speed with reference to data for calibration. In this case, the rotation speed of the sign can be calculated easily and quickly from the index value.

In yet another aspect of the invention, the object to be labeled is a nanoscale substance or field. In this case, nano-level fine labeling is required, but a relatively high diffraction signal can be obtained while ensuring fineness by labeling such as nanocrystals. In addition, a biomolecule or the like can be mentioned as a nanoscale substance, and a force field environment in a supersaturated network or the like can be mentioned as a nanoscale field.

It is a conceptual diagram explaining the motion measuring apparatus of 1st Embodiment. It is a conceptual diagram explaining the state of a sample. It is a conceptual diagram explaining the measurement state. It is a distribution map which shows the result of having calculated the standard deviation about the detection intensity of the diffraction from the label attached to this protein in the case of adding an activator and the case of adding an inhibitor to the protein to be measured. (A) shows the result of performing the same analysis as in FIG. 4 on the detection intensity of diffraction from the substrate, and (B) shows the same analysis as in FIG. 4 on the detection intensity of the background (scattered X-rays). The result is shown. (A) shows the result of measuring and analyzing the diffraction from the label by adding the activating substance to the protein to be measured, and (B) measuring the diffraction from the label by adding the inhibitor to the protein to be measured. The results of the analysis are shown. It is a conceptual diagram explaining the motion measuring apparatus of 3rd Embodiment.

[First Embodiment]
Hereinafter, the motion measuring device according to the first embodiment of the present invention will be described in detail with reference to FIG. 1 and the like.

The motion measuring device 100 of the first embodiment shown in FIG. 1 irradiates a sample with a label exhibiting a diffractive effect on X-rays with monochromatic X-rays, detects diffracted X-rays from the label, and diffracts the sample. This is to evaluate the motion state of a sample from the temporal fluctuation of X-rays and the temporal intensity change. The motion measuring device 100 includes a beam irradiation unit 20 that is an X-ray source, a sample stage 30 that supports a sample, a sensor 40 that captures a diffraction image, and a control device that comprehensively controls the operation of the motion measuring device 100 as a whole. 60 and.

The beam irradiating unit 20 irradiates a quantum beam on a nanocrystal or other label attached to an object such as a biomolecule. The beam irradiation unit 20 emits a single-color, beam-shaped irradiation X-ray L1 as a quantum beam with a desired intensity. As the irradiation X-ray L1, for example, Kα-ray can be used, but the irradiation X-ray is not limited to this. Further, the energy of the irradiation X-ray L1 is usually set appropriately in the range of about 10 KeV to 30 KeV according to the label and application, but is not limited to this. Although detailed description is omitted, the beam irradiation unit 20 has an electron beam excitation unit and an X-ray condensing unit. The former electron beam excitation section causes the accelerated electrons to enter the metal target, and utilizes, for example, linear spectral excitation X-rays generated at that time. The latter X-ray condensing unit is, for example, a tubular mirror member that utilizes total internal reflection, and temporarily converts X-rays from an electron beam excitation unit into a point light source. A filter for removing unnecessary wavelength components and a collimator for preventing the spread of the irradiated X-ray L1 can be added to the beam irradiation unit 20.

The sample stage 30 holds the sample cell 71 in a state of being set in an appropriate position. At this time, the sample cell 71 encloses an object such as a biomolecule, and is arranged in front of the irradiation X-ray L1 from the beam irradiation unit 20 in the emission direction. As a result, the target area of the sample cell 71 can be irradiated with the single-color irradiation X-ray L1. The sample stage 30 can be accompanied by a beam stopper that receives the irradiation X-ray L1 that passes through the sample cell 71 and travels straight.

The sensor 40 detects a quantum beam from a label attached to an object such as a biomolecule as an intense signal. The sensor 40 is a two-dimensional image sensor, and diffracted X-rays L2, which is a quantum beam generated by diffraction in the sample cell 71, are incident on the detection surface 41. The sensor 40 detects the intensity distribution of the X-ray diffraction pattern formed on the detection surface 41 as a two-dimensional signal intensity, that is, an image. The sensor 40 is a photon counting type detector that measures the intensity of light from a photon pulse, that is, an X-ray photon counting type two-dimensional detector. Specifically, PILATUS (pixel apparatus for the SLS) manufactured by Rigaku Co., Ltd. is used as the X-ray photon counting type two-dimensional detector. The X-ray photon counting type two-dimensional detector is a diode array having a structure in which a high-resistance silicon substrate is sandwiched between a common electrode to which a high voltage is applied and having an n-layer and a charge collecting electrode which is matrix-arranged and has a p-layer. This is a hybrid pixel type X-ray detector in which the diode array and the read integrated circuit are laminated. The intensity and pattern of the diffracted X-ray L2 can be read out as pixels by the sensor 40, and the change in the intensity of the diffracted X-ray L2 can be measured in pixel units by repeating the measurement with a fixed accumulation time. The sensor 40 is not limited to PILATUS, and may be a CMOS image sensor or a CCD image sensor.

The control device 60 controls the operation of the beam irradiating unit 20 via the light source driving unit 51, and can adjust the state such as the intensity of the monochromatic irradiation X-ray L1 emitted from the beam irradiating unit 20. The control device 60 controls the operation of the sample stage 30 via the stage drive unit 53, and can adjust the position and posture of the sample cell 71 supported by the sample stage 30. The control device 60 controls the operation of the sensor 40, extracts a necessary signal from the detection output of the sensor 40, and performs necessary processing on the extracted signal to exist in the sample cell 71. Analyze and evaluate the motor state of the subject.

The control device 60 has a structure similar to that of a general computer, and has an arithmetic processing unit, a storage unit, an image processing unit, an input / output unit, a communication unit, and the like. The control device 60 is a part that processes the detection output from the sensor 40 to acquire the motion state of an object such as a biomolecule, and functionally extracts a signal in a detection window set in the sensor 40, which will be described later. It is provided with a signal extraction unit 61 and a signal processing unit 62 that determines an index value based on the detection intensity of the detection window and evaluates the motion state of the marker and the motion state of the target based on the index value. The detection window for detecting the signal corresponds to a pixel provided on the detection surface 41 of the sensor 40.

The index value determined by the control device 60 can be determined by processing the signal intensity or the detection intensity from one specific pixel provided on the detection surface 41 of the sensor 40, but is provided on the detection surface 41. It can also be determined by processing the signal strength or the detection strength from a plurality of specific pixels in a complex manner. The signal from one or more pixels provided on the detection surface 41 is processed as a change in detection intensity with time, and the result is stored as an index value. This index value indicates a change in the detected intensity over time, more specifically, a temporal fluctuation or a temporal intensity change, and is used to estimate the rate of rotational motion of nanocrystals and other labels. Can be done.

FIG. 2 is a conceptual diagram showing a state of an object in the sample cell 71. In the sample cell 71, the target 74, which is a biomolecule or the like, is fixed on the substrate 73, which is one of the X-ray transmission windows, and nanocrystals, quantum dots, and other labels 75 are placed at specific points of the target 74. It is attached. For the method of fixing the target 74 to the substrate 73 and the method of chemically fixing the label 75 to the target 74, for example, "Picometer-scale dynamical observations of individual membrane proteins: The case of bacteriorhodopsin" Y. Okumura, T. Oka, M. The methods described in the literature such as Kataoka, and YC Sasaki, Phys. Rev. E70, 021917 (2004) can be used. In this case, between the substrate 73 and the biomolecule (object 74), the amino group of the biomolecule is specifically fixed to the substrate via SPDP (3- (2-pyridyldithio) propionic acid N-succinimidyl). Will be done. Further, between the biomolecule and the gold crystal (label 75), a direct bond between the thiol group of the cysteine residue of the biomolecule and gold is performed.

The marker 75 rotates or translates with the movement of the subject 74. When the portion of the target 74 to which the marker 75 is attached rotates, the indicator 75 also rotates in the same manner. When the marker 75 is rotated in this way, the spot-shaped diffracted X-ray L2 by the marker 75 is also rotated. Therefore, if the motion of the diffracted X-ray L2 can be followed, the motion of the specific portion of the target 74 can be captured. However, when the diffracted X-ray L2 is monochromatic, the diffracted X-ray L2 appears as a bright spot blinking at one point on the diffracted ring peculiar to the nanocrystal as the label 75, and is therefore regarded as the locus of the diffracted X-ray L2. It is difficult. Therefore, the bright spots that blink on such a diffraction ring are measured by one or more pixels provided on the detection surface 41 of FIG. 1, and the detection intensity is measured by a change over time (specifically, time) in the detection intensity. By converting it into information indicating fluctuations and changes in intensity over time, it is possible to estimate the motion (that is, the rotational speed) of a specific location of the marker 75 or the target 74.

The label 75 is, for example, a gold nanocrystal or the like, and is formed by epitaxial growth or the like. The diameter size of the gold nanocrystals is, for example, about 20 to 40 nm, but it can also be about 2 to 5 nm.

A plurality of objects 74 can be fixed to the substrate 73. In this case, at the position of the corresponding diffraction ring on the detection surface 41, a plurality of blinking bright spots corresponding to the plurality of labels 75 attached to the plurality of objects 74 are detected. If these are collectively processed, an index relating to the rotational speed of the specific location of the target 74 obtained by averaging the rotational movements of the plurality of markers 75 can be obtained.

With reference to FIG. 3, a specific method for calculating the signal strength from the pixels provided on the detection surface 41 or the index value using the detection strength will be described. FIG. 3 shows the detection surface 41 of the sensor 40, and the detection surface 41 extends parallel to the xy surface. The arc centered on the origin O shows a part of the virtual diffraction ring R1 corresponding to the nanocrystal which is the label 75. Specifically, the diffraction ring R1 corresponds to the (111) plane of gold when the label 75 is, for example, a gold nanocrystal. A vertically long and rectangular detection region A1 is set at one location along the diffraction ring R1. As shown in an enlarged manner in the figure, the detection region A1 includes a plurality of pixels P1 as a plurality of detection windows, and is a region defined by the plurality of pixels P1. In the detection region A1, the blinking pattern FP of the diffracted X-ray L2 is formed. The blinking pattern FP is point-like, but usually moves slightly depending on the monochromaticity of X-rays and shows a locus tr.

The signal extraction unit 61 of FIG. 1 gradually extracts the detection intensity measured by the plurality of pixels (detection windows) P1 arranged in the detection area A1 of the detection surface 41 of the sensor 40 as data, and the signal processing unit 62 gradually extracts the detection intensity. , The detection intensity extracted by the signal extraction unit 61 is accumulated and converted into information on changes over time. Specifically, the signal processing unit 62 statistically evaluates, for example, the temporal fluctuation of the detection intensity. The detection intensity for each frame obtained from each pixel P1 in the detection region A1 is accumulated over time by a plurality of extractions, and is converted into, for example, a standard deviation (SD). Here, it is desirable that the exposure time of one frame measured on the detection surface 41 or the pixel P1 is appropriately set according to conditions such as the motion speed of the target 74. The exposure time for detection can be set in the range of, for example, 1 second to several tens of seconds. Further, the number of extractions of the detection intensity in each pixel P1 is, for example, 30 times or more, more preferably 100 times or more, from the viewpoint of ensuring statistical reliability. In this case, a standard deviation (SD) is obtained for each pixel P1 with respect to the detection intensity. By increasing the number of pixels P1 included in the detection region A1, the statistical reliability in estimating the motion velocity can be improved.

FIG. 4 shows the results of detecting and analyzing the diffraction from the label 75 when the label 75 was attached to the protein to be measured 74 and the activator was added and the inhibitor was added. It is a distribution map. This distribution map shows the result of calculating the standard deviation (SD) for the detection intensity for each pixel (detection window) P1 in the detection area A1 where the diffraction image was taken, and the horizontal axis is the standard deviation (SD). Yes, the vertical axis shows the frequency. The frequency on the vertical axis indicates the number of pixels indicating the standard deviation on the horizontal axis. As the subject 74, an acetylcholine binding protein (AChBP) is used here. In addition, acetylcholine and Ach are used as activators or agonists for proteins, and α-bungarotoxin and Bgtx are used as inhibitors or antagonists. The protein of interest 74 is immobilized on a film of Kapton ™, which is a polyimide substrate 73, and is labeled with a label 75, which is a gold nanocrystal. “Ach_2s” in the chart means that the acetylcholine-binding protein labeled with gold nanocrystals was added with acetylcholine, and the change over time was measured by exposure for 2 seconds, and “Bgtx_2s” means acetylcholine labeled with gold nanocrystals. It means that α-bungarotoxin was added to the bound protein and the change over time was measured by exposure for 2 seconds. As is clear from the figure, the standard deviation, that is, the variation is smaller overall when Ach is added than when Bgtx is added, and the peak positions of both are clearly different between 190 and 220, respectively. It is happening. That is, it is judged that the labeled 75 of the protein to which Ach is added has a smaller standard deviation, that is, the variation of the detected values than the labeled 75 of the protein to which Bgtx is added, and is exercising at a relatively high speed. Therefore, by observing the marker 75 with a plurality of pixels P1 and obtaining the standard deviation of the intensity thereof, the rotation speed of the marker 75 and the rotation speed of the specific portion of the target 74 can be estimated using this standard deviation as an index. Specifically, if the conversion table and the calibration curve are prepared in advance in the conversion unit 62a provided in the signal processing unit 62 according to the parameters for specifying the target 74 and the label 75 and specifying other measurement conditions, the above The rotation speed of a specific point of the target 74 can be determined from the standard deviation such as. In the above, the rotation speed is estimated from the value of the standard deviation, but it is also possible to estimate the rotation speed from the peak value of the count value related to the standard deviation in FIG.

In the above, from the viewpoint of ensuring statistical reliability, the standard deviation was calculated based on the detection intensities from a plurality of pixels P1, but the standard deviation is calculated based on the detection intensities from a single pixel P1. It is also possible, and the rotation speed of the specific portion of the target 74 can be determined from such a standard deviation.

FIG. 5A shows the results of the same measurement as in FIG. 4 for the substrate 73 of Kapton ™ to which the target 74 is added, using the diffraction ring derived from Kapton as a detection window for reference. In this case, it can be seen that there is almost no difference in the signal at the diffraction position when Ach or Bgtx is added. Further, FIG. 5B shows the results of the same measurement as in FIG. 4 for the Kapton substrate 73 to which the target 74 is added, with the portion where the diffraction ring derived from a specific substance cannot be seen as the detection window for reference. Is shown. In this case, it can be seen that there is no difference in the signal at the diffraction position when Ach or Bgtx is added. As is clear from comparing these FIGS. 5 (A) and 5 (B) with FIG. 4, the label 75 is measured by the pixel P1 and the standard deviation (SD) is obtained from the detected intensity of the label 75. The exercise speed of a specific part of the target 74 can be estimated.

FIG. 6A shows the result of measuring the diffraction X-ray from the label 75 in pixel units of the detection surface 41 using AChBP to which Ach is added as the target 74, and FIG. 6B shows Bgtx as the target 74. The result of measuring the diffracted X-ray from the label 75 in the pixel unit of the detection surface 41 by using AChBP which added the above is shown. However, the horizontal axis is the exposure time, and the vertical axis is the standard deviation. In this case, the standard deviation of the detection intensity is calculated while changing the exposure time. In both graphs, the slope of the standard deviation of the detection intensity derived from Kapton is shown by the dotted line.

In the case of AChBP + Ach labeled with gold nanocrystals, the slope of the standard deviation graph by fitting is -0.46 (see FIG. 6 (A)), and in the case of AChBP + Bgtx labeled with gold nanocrystals 75, the slope is -0.46. The slope of the standard deviation graph by fitting is -0.45 (see FIG. 6B). On the other hand, in the case of Kapton, the slope of the standard deviation graph by fitting is −0.50 (see FIGS. 6 (A) and 6 (B)). The slope of Kapton is the same as that of the non-moving object, and the AChBP with the addition of Ach and Bgtx with motion shows different behavior from the non-moving object. There is no big difference between the slope of the AchBP graph with Ach added and the slope of the AChBP graph with Bgtx added, but from the slope of the standard deviation graph with varying exposure times, the markers 75 and 74 It may be possible to estimate the presence or absence of exercise at a specific location and the exercise speed.

As described above, in the motion measuring device 100 of the first embodiment, the standard deviation as an index value is determined based on the change over time in the detection intensity of the signal corresponding to the diffraction point by the predetermined pixel (detection window) P1. Since it is determined, it is not necessary to capture the motion and trajectory of the diffraction point over a wide range, and it is possible to measure using a simple device that generates quantum rays in a single color or a narrow wavelength range as a quantum radiation source and has a small irradiation flux. .. Further, the standard deviation as an index value determined based on the change over time of the detection intensity by the pixel (detection window) P1 indirectly represents the movement of the diffraction point and the motion speed of the marker 75. Therefore, it is possible to appropriately evaluate the motion state of the marker based on such an index value.

[Second Embodiment]
Hereinafter, the motion measuring device according to the second embodiment will be described. The motion measuring device according to the second embodiment is a modification of the first embodiment, and is the same as the first embodiment except for a portion not particularly described.

In the case of the motion measuring device 100 of the second embodiment, the detection intensity of the diffracted X-ray L2 or the rate of change of the signal, specifically, the differential value is used as an index for evaluating the motion state of the marker 75. Here, the differential value of the detected intensity includes an absolute value and a relative value. Time series information is not used in the analysis using simple standard deviation. Therefore, it is considered that the detected intensity or the differential value of the signal is more suitable for extracting the kinetic component. The differential value of the detected intensity means the amount of change in the detected intensity from each pixel P1 acquired in time series for each time unit, for example, one frame. Among the methods using the differential value, the one using the absolute value is a simple difference between the detection intensity of a certain pixel P1 and the output intensity of the same pixel P1 of the previous frame. On the other hand, among the methods using the differential value, the method using the relative value means the rate of change (%) between the detection intensity of a certain pixel P1 and the output intensity of the same pixel P1 of the previous frame. When comparing different samples, it is considered that the comparison of the differential value as a relative value is more effective than the comparison of the differential value as an absolute value. That is, if the relative differential value is calculated, it is possible to make an equivalent comparison between the same sample or different samples without performing calibration or the like. In the case of comparison between different sample systems, the value of the signal intensity obtained on the sensor 40 may change significantly due to the difference in X-ray absorption (scattering) rate due to the difference in solvent, and the absolute differential value may be used. When used, it tends to be difficult to compare the degree of change.

[Third Embodiment]
Hereinafter, the motion measuring device according to the third embodiment will be described. The motion measuring device according to the third embodiment is a modification of the first embodiment, and is the same as the first embodiment except for a portion not particularly described.

As shown in FIG. 7, in the case of the motion measuring device of the third embodiment, two detection regions A1 and A2 are provided along the diffraction ring R1 on the detection surface 41 of the sensor 40. One detection region A1 extends in the y direction, and the other detection region A2 extends in the x direction. In this case, the anisotropy of the motion of the label 75 can be detected. Specifically, one detection region A1 can estimate the movement of the diffracted X-ray L2 by the label 75 in the x direction, and the other detection region A2 can estimate the movement of the diffracted X-ray L2 by the label 75 in the y direction. Can be done. That is, there is a possibility that the rotation around the x-axis and the rotation around the y-axis can be detected separately with respect to the movement of the marker 75 or the target 74 at a specific location. However, in the case of performing such measurement, when a plurality of objects 74 are fixed on the substrate 73 of the sample cell 71, it is significant data that the directions of these objects 74 are aligned and oriented. It becomes a premise for.

〔others〕
Although the present invention has been described above according to the embodiment, the present invention is not limited to the above embodiment.

For example, an electron beam or a neutron beam can be used as the quantum beam. In this case, the beam irradiation unit 20 irradiates the sample cell 71 with an electron beam or a neutron beam, and the diffraction line from the sample cell 71 is detected by the sensor 40. As the sensor 40, an electronic counting type two-dimensional detector or a neutron counting type two-dimensional detector can be used, but a CMOS image sensor or a CCD image sensor can also be used. Further, as the label 75 when an electron beam is used, for example, a gold colloid can be used, and as the label 75 when a neutron beam is used, for example, nanodiamond can be used.

In the above, a group of pixels P1 are adjacent to each other in the detection region A1, but the standard deviation or the like can be calculated based on the detection intensities from a plurality of pixels P1 that are separated from each other.

In the above, it is assumed that the observation target 74 is a biomolecule or the like, but the motion of various nanoscale substances can be measured, not limited to the biomolecule. That is, a polymer polymer or other inorganic material can be an observation target 74, and the physical properties of such an inorganic material can be evaluated. In a special case, the dynamics of an object in a solution of nanobubbles (nano-sized very stable bubbles) can also be measured. It is also possible to check the quality of nanoscale structures (nanomachines). Regarding the quality check of the nanomachine, if it is a defective product, it is considered that the fluctuation of the diffraction image becomes large.

Further, not only the substance itself but also a nanoscale environment (for example, a force field) can be measured. In the case of measurement of the force field generated by the formation and collapse of a supersaturated network in a nanoscale environment, for example, a supersaturated solution, the label 75 such as nanocrystals is dispersed in the supersaturated solution. In this case, the label 75, which is a nanocrystal or the like, moves around in the solution under the force from the supersaturated network, and by evaluating such motion, it is possible to capture the force field or the like in the nanoscale environment. In another example, it is also possible to observe the motion of nanocrystals floating in the solution being repelled by the radiation pressure generated by high-intensity laser light irradiation. It should be noted that the case of measuring the nanoscale environment in this way is also included in the concept of the motion measuring device.

In the above, the diffraction image from the sample cell 71 is collectively photographed by the sensor 40, but the sensor 40 is miniaturized and the small sensor is moved to an arbitrary place corresponding to the detection area A1 or the like by the drive stage. You can also do it.

20 ... Beam irradiation unit, 30 ... Sample stage, 40 ... Sensor, 41 ... Detection surface, 51 ... Light source drive unit, 53 ... Stage drive unit, 60 ... Control device, 61 ... Signal extraction unit, 62 ... Signal processing unit, 71 ... sample cell, 73 ... substrate, 74 ... target, 75 ... label, 76 ... reference marker, 100 ... motion measuring device, A1, A2 ... detection area, FP ... blinking pattern, L1 ... irradiation X-ray, L2 ... diffracted X-ray , P1 ... pixel, R1 ... diffraction ring

Claims (10)

A beam irradiating unit that irradiates a quantum beam on the sign attached to the target,
A sensor that detects the quantum beam from the label and
A signal extraction unit that extracts a signal in a predetermined detection window set in the sensor, and a signal extraction unit.
It is provided with a signal processing unit that determines an index value based on the change over time in the detection intensity of the signal by the predetermined detection window and evaluates the motion state of the marker based on the determined index value.
In the sensor, a detection area defined by one pixel or a plurality of pixels on the detection surface is set.
The detection area is a motion measuring device set to be arranged and sized according to the characteristics of the marker. The motion measuring device according to claim 1, wherein the sensor detects a quantum beam from the marker as an image. It said predetermined detection window is pixels arranged in a matrix in the detection surface of the sensor, movement measurement device according to any one of claims 1 and 2. A beam irradiating unit that irradiates a quantum beam on the sign attached to the target,
A sensor that detects the quantum beam from the label and
A signal extraction unit that extracts a signal in a predetermined detection window set in the sensor, and a signal extraction unit.
It is provided with a signal processing unit that determines an index value based on the change over time in the detection intensity of the signal by the predetermined detection window and evaluates the motion state of the marker based on the determined index value.
The index value is a motion measuring device relating to the variation in the change of the detected intensity with time. The motion measuring device according to claim 4, wherein the index value is a standard deviation. A beam irradiating unit that irradiates a quantum beam on the sign attached to the target,
A sensor that detects the quantum beam from the label and
A signal extraction unit that extracts a signal in a predetermined detection window set in the sensor, and a signal extraction unit.
It is provided with a signal processing unit that determines an index value based on the change over time in the detection intensity of the signal by the predetermined detection window and evaluates the motion state of the marker based on the determined index value.
The beam irradiation unit irradiates a monochromatic X-ray as a quantum beam.
The sensor is a motion measuring device that detects X-rays diffracted by the sign. The motion measuring device according to any one of claims 1 to 6 , wherein the label is a nanocrystal. A beam irradiating unit that irradiates a quantum beam on the sign attached to the target,
A sensor that detects the quantum beam from the label and
A signal extraction unit that extracts a signal in a predetermined detection window set in the sensor, and a signal extraction unit.
It is provided with a signal processing unit that determines an index value based on the change over time in the detection intensity of the signal by the predetermined detection window and evaluates the motion state of the marker based on the determined index value.
A motion measuring device that evaluates the rotational speed of the marker as the motion state of the marker. A beam irradiating unit that irradiates a quantum beam on the sign attached to the target,
A sensor that detects the quantum beam from the label and
A signal extraction unit that extracts a signal in a predetermined detection window set in the sensor, and a signal extraction unit.
It is provided with a signal processing unit that determines an index value based on the change over time in the detection intensity of the signal by the predetermined detection window and evaluates the motion state of the marker based on the determined index value.
The signal processing unit is a motion measuring device having a conversion unit that converts an index value into a rotation speed with reference to calibration data. The motion measuring device according to any one of claims 1 to 9 , wherein the object to be labeled is a nanoscale substance or a field.

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