Patent Grant
10838188
An aspect of the present invention relates to an optical tweezers device.
The optical tweezers technology is known which is a technology for trapping, for example, a particle measuring about 1 μm and, furthermore, moving it (refer to Patent document 1, for example). According to the optical tweezers technology, a laser beam is focused with a lens and a particle that is brought close to a focusing point is trapped by optical pressure acting on it. The optical tweezers technology makes it possible to continue to trap a particle by, for example, causing the particle to exist in a liquid, producing a refractive index difference between the particle and liquid around it, and directing the total force of optical pressure acting on the particle to the focusing point.
To trap a particle utilizing the optical tweezers technology described above, it is necessary that the particle have transmittivity (transmit the laser beam) and that the refractive index (n2) of the particle is larger than the refractive index (n1) of liquid around it (n2>n1).
Patent Document 1: JP-A-2006-235319
In optical tweezers devices of the above kind, a particle trapped by a laser beam can be moved by moving the focusing point of the laser beam (the focal point of the lens). This may be done by, for example, translating the laser beam by combining plural acousto-optic modulators. However, this results in problems that the overall device becomes expensive and a very wide space is necessary to install component devices.
In view of the above, a mechanism has been proposed that, instead of moving the focusing point of the laser beam, moves a stage that is mounted with a holding member such as a prepared slide that holds a particle to be trapped and thereby moves what surrounds the particle being trapped by the laser beam relative to the particle.
In an optical tweezers device having such a mechanism, when a particle being trapped is imaged by an imaging means (camera), motion of the particle (a movement relative to its neighborhood) is output on a monitor as, for example, a moving image, and an operator observes the moving image, the operator should see that an untrapped particle etc. that exist around the particle being trapped move whereas the particle being trapped is displayed on the screen so as not to move.
Thus, the operator cannot follow a movement of the particle being trapped. Even if a movement of the particle is observed by watching a moving image, there is no clear reference position and a movement locus of the particle is unknown. If there exists an untrapped particle around the particle being trapped, a relative movement of the particle being trapped can be recognized because the untrapped particle moves with respect to the particle being trapped. However, if there does not exist any other particle (not to be trapped) around the particle being trapped, it is unknown at all whether the particle being trapped is moving.
As such, optical tweezers devices that employ the mechanism of moving the stage while fixing the focusing point of the laser beam cannot be used suitably as, for example, a measurement tool that traps and moves a particle.
An object of an aspect of the invention is therefore to make it possible to perform processing so as to produce an image in which a particle being trapped appears as if it were moving even in an optical tweezers device that employs a mechanism of moving a stage while fixing the focusing point of the laser beam.
To attain the above object, an optical tweezers device according to an aspect of the invention includes: a light source which emits a laser beam; a lens which focuses the laser beam emitted from the light source; a device base on which the light source and the lens are mounted; a stage which is disposed on the device base so as to be movable and to be able to support a holding member which holds a particle to be trapped by the focused laser beam; a drive unit which moves the stage; an acquisition unit which acquires basic movement information indicating a movement of the stage caused by the drive unit; a conversion unit which converts the movement of the stage indicated by the basic movement information into a movement of the particle being trapped and generates converted movement information indicating the movement of the particle; and a distance calculation unit which determines a distance between the particle being trapped by the laser beam and a focal point of the lens, wherein the conversion unit generates the converted movement information by correcting the movement of the particle using the distance determined by the distance calculation unit.
In the configuration according to the aspect of the invention, the focusing point of the laser beam is fixed and the stage is moved. In actuality, things other than the particle being trapped are moved relative to the particle. However, processing can be performed so as to produce a virtual movement of the particle by converting the movement of the stage into a movement of the particle being trapped. As the holding member which is mounted on the stage moves, the particle being trapped by the laser beam follows the focal point of the lens. However, there may occur a case that the center of the particle does not coincide with the focal point of the lens. In view of this, a distance between the particle being trapped and the focal point of the lens is determined. And the movement of the particle is corrected using the determined distance and converted movement information indicating a movement of the particle is generated. This makes it possible to output, accurately, how the particle moves. In this manner, with the mechanism that the focusing point of the laser beam is fixed and the stage is moved, it becomes possible to perform processing so as to produce a virtual movement of a particle being trapped by the laser beam.
The optical tweezers device may further include: an imaging unit which acquires an image of a region, including the particle and its neighborhood, in the holding member; and an output unit which outputs a virtual image in which the particle being trapped appears as if it were moving on the basis of the converted movement information generated by the conversion unit.
This allows an operator to recognize, intuitively, a movement locus, for example, of the particle as a manner of its movement.
The optical tweezers device may further include a strain gauge which detects a movement distance of the stage, the drive unit has a mechanism that uses deformation of the piezoelectric element as motive power for moving the stage, and the acquisition unit determines the movement distance of the stage to be included in the basic movement information on the basis of a detection signal of the strain gauge.
In this case, a movement of the stage can be detected accurately by the strain gauge and the stage can be moved at a high resolution by the mechanism including the piezoelectric element.
The aspect of the invention makes it possible to perform processing so as to produce and image in which a particle being trapped appears as if it were moving by the laser beam were moving even in an optical tweezers device that employs a mechanism of moving a stage while fixing the focusing point of the laser beam.
An embodiment of the present invention will be hereinafter described on the basis of the drawings
As described later, the optical tweezers device 1 is configured in such a manner that the stage 46 can be moved by the drive means 48 with respect to the device base 45 which is fixed to a working floor. And other devices, that is, the light source 10, the lenses 28 and 31, the detector 40, the imaging means 50, etc., are fixed to the device base 45 and are not moved with respect to the device base 45.
The light source 10 of the laser beam, which is a laser device for emitting the laser beam L, emits the laser beam L having a first wavelength according to a control signal received from the control means 60. A particle being held by a holding member (e.g., prepared slide) 47 mounted on the stage 46 is trapped by an optical tweezers technique using the laser beam L (optical trap).
The light guiding means (21-27) serve to guide the laser beam L emitted from the light source 10 to the first lens 28. The light guiding means (21-27) will be described below in order.
A first reflection mirror 21 reflects the laser beam L coming from the light source 10 so that it is incident on a first aperture 22. The first aperture 22 narrows the diameter of the incident laser beam L and outputs resulting light toward a first collimating lens 23. The first collimating lens 23 enlarges the diameter of the laser beam L and outputs resulting light toward a second collimating lens 24. The second collimating lens 24 converts the diameter-enlarged laser beam L into parallel light and outputs it to a second aperture 25. The second aperture 25 narrows the diameter of the parallel laser beam L and outputs resulting light toward a first mirror 26. The first mirror 26 reflects the incident laser beam L toward a second mirror 27. The second mirror 27 reflects the incident laser beam L toward the first lens 28.
The first lens 28 focuses the laser beam L coming from the second mirror 27 at a focal point that is set in the holding member 47. A particle that is brought close to the focusing point (the focal point of the lens 28) can be trapped by the laser beam L focused by the lens 28. The focused laser beam L is incident on the second lens 31 after passing through the particle. The laser beam L incident on the second lens 31 after passing through the particle is output toward the third mirror 33, is reflected by the third mirror 33, and is incident on the detector 40. The mirrors 33 and 26 transmit illumination light S coming from the illumination light source 30.
The illumination light source 30, which is, for example, an LED illumination device, emits illumination light S having a second wavelength according to a control signal received from the control means 60. The illumination light S serves as illumination light for the imaging means 50 for observing a state of a particle being held by the holding member 47. The illumination light S passes through the third mirror 33, is focused by the second lens 31, thereafter passes through the first lens 28, is reflected by the second mirror 27, passes through the first mirror 26, and reaches the imaging means 50.
The holding member 47 for holding a particle is mounted on the stage 46. A fluid W and a particle C1 to be trapped that is contained in the fluid W are held by the holding member 47 (see
As described above, the holding member 47 moves together with the stage 46 relative to the lens 28, whereby the particles C2 not to be trapped move relative to the particle C1 to be trapped.
The stage 46 is supported so as to be movable in the front-rear direction, the left-right direction, and the top-bottom direction, and the drive means 48 moves the stage 46 in the front-rear direction, the left-right direction, and the top-bottom direction. In
The optical tweezers device 1 is also equipped with sensors for detecting a movement of the stage 46. In the embodiment, strain gauges, that is, a strain gauge 49a for detecting a movement distance of the stage 46 in the X-axis direction, a strain gauge 49b for detecting a movement distance of the stage 46 in the Y-axis direction, a strain gauge 49c for detecting a movement distance of the stage 46 in the Z-axis direction, are provided as the sensors.
Detection signals (voltage signals) that are output from the strain gauges 49a, 49b, and 49c are amplified by an amplifier 44 and collected by a data accumulation unit (DAQ) 43 which functions in cooperation with the control means 60 which is a computer.
The following description will be made of a case that the stage 46 is moved toward the positive side in the Y-axis direction in the XY plane (see
Referring to
Since as mentioned above the holding member 47 including the particle C1 being trapped is moved toward the positive side in the Y-axis direction with respect to the lens 28 (see
Thus, the laser beam L that reaches the photodetection units A1, A2, A3, and A4 after being emitted from the light source 10 and passing through the particle C1 being trapped is detected at a position that is deviated in the Y-axis direction from the reference position N (see
The distance d0 (V) means the distance d (m) (see
The imaging means 50 shown in
The control means 60, which is, for example, a computer that is equipped with a processor and a memory, is provided with the above-mentioned function of outputting control signals, a function of processing each of pieces of information (each of signals) collected by the data accumulation unit 43, a function of taking in image information from the imaging unit 50 and generating new image information, and a function of outputting various kinds of information. The control means 60 is also equipped with the distance calculation unit 61, an acquisition unit 62, a conversion unit 63, and an image generation unit 64 as function units that are implemented by the processor's running computer programs that are stored in the memory of the computer. These function units can process each of pieces of information (each of signals) collected by the data accumulation unit 43. The optical tweezers device 1 according to the embodiment is further equipped with a monitor (output means) 70 for outputting, as an image, image information generated through processing by the control means 60.
The distance calculation unit 61 determines, through calculation, a distance d (see
The acquisition unit 62 can determine a movement distance of the stage 46 in the X-axis direction on the basis of a detection signal received from the strain gauge 49a whose detection direction is the X-axis direction, and can determine a movement distance of the stage 46 in the Y-axis direction on the basis of a detection signal received from the strain gauge 49b whose detection direction is the Y-axis direction. The acquisition unit 62 discriminates between the X-axis direction and the Y-axis direction for movement of the sage 46 by acquiring detection signals of the two strain gauges 49a and 49b with discrimination between them. Thus, the acquisition unit 62 acquires a movement direction and a movement distance in that movement direction in such a manner that they are correlated with each other. In this manner, the acquisition unit 62 can acquire basic movement information indicating a direction and a distance of a movement of the stage 46 caused by the drive means 48.
The basic movement information includes information indicating a movement direction of the stage 46 and information indicating a movement distance in that movement direction. In the embodiment, since as mentioned above the stage 46 is moved with respect to the lens 28 in a direction that is parallel with the XY plane, the conversion unit 63 performs processing of converting the movement direction of the stage 46 and the movement distance in that movement direction into a movement direction of the focusing point of the lens 28 (or the laser beam) and a movement distance in that movement direction on the basis of the basic movement information.
More specifically, in the embodiment, in actuality, the stage 46 has been moved toward the positive side in the Y-axis direction by a movement distance Δy with respect to the fixed focusing point of the lens 28 (or the laser beam) and this manner of movement has been acquired by the acquisition unit 62 as the basic movement information. Thus, the conversion unit 63 generates, on the basis of the basic movement information, information (hereinafter referred to as “pre-correction movement information”) indicating that the focusing point of the lens 28 (or the laser beam) has been moved virtually with respect to the stage 46 by the movement distance Δy toward the negative side in the Y-axis direction. That is, the conversion unit 63 generates pre-correction movement information indicating that the focusing point of the lens 28 (or the laser beam) has been moved by the same movement distance indicated by the basic movement information in the direction opposite to the movement direction indicated by the basic movement information.
In actuality, although as shown in
On the other hand, the generated pre-correction movement information is information indicating the direction and the distance of the movement of the focusing point of the lens 28 (or the laser beam) with respect to the stage 46. Thus, if the movement distance itself of the focusing point of the lens 28 (or the laser beam) indicated by the pre-correction movement information is regarded as a movement distance of the particle C1, there occurs a difference between the center position of the particle C1 and the focal point Q of the lens 28, that is, an error in the above-mentioned distance d.
In view of the above, the conversion unit 63 corrects the movement distance contained in the pre-correction movement information and generates converted movement information indicating a movement direction of the particle C1 and a corrected movement distance in that movement direction. The information indicating the movement direction contained in the converted movement information is the same as that contained in the pre-correction movement information.
The above correction is performed using the distance d determined by the distance calculation unit 61. More specifically, since as shown in
As described above, the conversion unit 63 converts, virtually, a movement of the stage 46 indicated by the basic movement information into a movement of the particle C1 being trapped and generates converted movement information indicating the movement of the particle C1. In doing so, the conversion unit 63 corrects the movement of the particle C1 being trapped using the distance d determined by the distance calculation unit 61 and thereby generates converted movement information indicating the movement direction of the particle C1 and a movement distance in that movement direction.
In the optical tweezers device 1 according to the embodiment which, as mentioned above, is equipped with the imaging means (camera) 50, an image including the particle C1 being trapped and its neighborhood is acquired and information of that image is acquired by the control means 60.
The image generation unit 64 which is one of the function units of the control means 60 generates converted image information for output on the monitor 70 on the basis of the image information acquired by the image means 50 and the converted movement information generated by the conversion unit 63. An image obtained from this image information is an image including the particle C1 being trapped and its neighborhood, and is, for example, a virtual image that is a combination of a background image that is an image that was acquired by the imaging means 50 before the movement and from which the particle C1 being trapped is excluded and a manner of movement of the particle C1 that is obtained from the converted movement information.
If the above-described conversion processing were not performed, an image (moving image) that is obtained when the stage 46 is moved being driven by the drive means 48 would be such that as shown in
In contrast, in the embodiment, as described above, the conversion unit 63 generates converted movement information. On the basis of the converted movement information, the image generation unit 64 of the control means 60 generates virtual image information (moving image information) by converting the movement of the stage 46 into a movement of the particle C1 being trapped. The control means 60 causes the monitor 70 to output this image information as shown in
More specifically, an image is output in which the particle C1 being trapped moves in the Y-axis direction with respect to its neighborhood (because the movement direction information contained in the converted movement information is information indicating the Y-axis direction) by a movement distance (Δy−d) (because the information of the movement distance, contained in the converted movement information, in the above movement direction is information indicating a movement distance (Δy−d) as corrected by a distance d).
The symbols, the arrows indicating the movements of the particles, the virtual lines (two-dot chain lines) indicating the particles before the movements, etc. that are shown in
The optical tweezers device 1 performs controls for causing the above-described pieces of processing, that is, the detection of a distance d0 (see
Although the optical tweezers device 1 according to the embodiment is configured as described above, the light guiding means (21-27) shown in
The distance calculation unit 61 of the control means 60 determines a distance d between a particle C1 being trapped by the laser beam L and the focal point Q of the lens 28. The acquisition unit 62 of the control means 60 acquires basic movement information indicating a movement (movement distance and movement direction) caused by the drive means 48, and the conversion unit 63 converts the movement of the stage 46 indicated by the basic movement information into a movement of the particle C1 being trapped and thereby generates converted movement information indicating the movement of the particle C1. The converted movement information generated by the conversion unit 63 indicates a movement of the particle C1 being trapped as corrected using the distance d determined by the distance calculation unit 61.
As described above, the optical tweezers device 1 according to the embodiment is configured in such a manner that the focusing point of the laser beam L (the focal point Q of the lens 28) is fixed and the stage 46 is moved. In actuality, things other than the particle C1 being trapped are moved relative to the particle C1. However, processing is performed so as to produce a virtual movement of the particle C1, that is, the functions of the control means 60 converts the movement of the stage 46 into a movement of the particle C1 being trapped by the laser beam L.
As the holding member 47 which is mounted on the stage 46 moves, the particle C1 being trapped by the laser beam L follows the focal point Q of the lens 28. However, there may occur a case that the center of the particle C1 does not coincide with the focal point Q of the lens 28. In view of this, in the embodiment, a distance d between the particle C1 being trapped and the focal point Q of the lens 28 is determined. The movement of the particle C1 is corrected using the distance d and converted movement information indicating a movement of the particle C1 is generated. This makes it possible to output, accurately, how the particle C1 moves. More specifically, a movement (locus) of the particle C1 being trapped can be visualized.
More specifically, the imaging means 50 acquires an image of a region, including the particle C1 and its neighborhood, in the holding member 47. The monitor 70 outputs a virtual image in which the particle C1 being trapped appears as if it were moving (see
For observation through the monitor 70, the image generation unit 64 makes a setting that a prescribed position of an image displayed on the monitor 70 (e.g., the center of the monitor 70) is made a reference point. It is preferable that the image generation unit 64 generate image information in which this reference point is fixed. This makes it easier to control a movement of the stage 46 so that the particle C1 being trapped is returned to its original position in an image. The movement can be facilitated also in a case that the stage 46 is moved manually. For example, where the center of the particle C1 being trapped (the focal point Q of the lens 28) is set coincident with the image center of the monitor 70 (reference point) at the beginning, the particle C1 can be returned to the reference point easily even after it has been moved randomly (in actuality, the stage 46 has been moved parallel with the XY plane).
In the embodiment, as described above, the drive means 48 for moving the stage 46 has the piezoelectric elements 48a, 48b, and 48c and employs the mechanism that uses deformation of each of the piezoelectric elements 48a, 48b, and 48c to produce motive power for moving the stage 46. The optical tweezers device 1 is further equipped with the strain gauges 49a, 49b, and 49c for detecting a movement distance of the stage 46. The acquisition unit 62 which is one of the function units of the control means 60 determines a movement distance of the stage 46 to be included in the basic movement information on the basis of detection signals of the strain gauges 49a, 49b, and 49c. This configuration makes it possible to control a movement of the stage 46 at a high resolution by performing a feedback control using detection signals of the strain gauges 49a, 49b, and 49c.
The embodiment disclosed above is illustrative and is not restrictive in every point. That is, the optical tweezers device according to the invention is not limited to the one in the illustrated embodiment and may be one in any other embodiment within the scope of the invention.
The movement distance information in each of the basic movement information, pre-correction movement information, and converted movement information may be a dimension value, it may be the number of pixels so as to be compatible with information handled in the imaging means 50 and the monitor 70.
Although the above embodiment has been described for the case that a manner of movement of the particle C1 is output on the monitor 70 in the form of a moving image on the basis of converted movement information, it may be output in a manner in which another means is used; it may be output in the form of a graph or numerical values. The above-described optical tweezers device 1 can be applied to various kinds of measurements and observations, and can be applied to a flow velocity measurement in micro-PIV (particle image velocimetry).
The present application is based on Japanese Patent Application No. 2015-207923 filed on Oct. 22, 2015, the disclosure of which is incorporated herein by reference.
| Number | Date | Country | Kind |
|---|---|---|---|
| 2015-207923 | Oct 2015 | JP | national |
| Filing Document | Filing Date | Country | Kind |
|---|---|---|---|
| PCT/JP2016/081225 | 10/21/2016 | WO | 00 |
| Publishing Document | Publishing Date | Country | Kind |
|---|---|---|---|
| WO2017/069228 | 4/27/2017 | WO | A |
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