JP2004354345A - Biomolecule analysis apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a biomolecule analysis apparatus which performs a multi-point fluorescence correlation analysis while immobilizing an object to be observed by optical trapping. <P>SOLUTION: The biomolecule analysis apparatus for analyzing dynamic behavior of biomolecules comprises a holding means (20) which optically holds the biomolecules in a measurable state, an image acquisition means (35) which acquires an image corresponding to at least one observation area of a biomolecular sample (S) containing the biomolecules held by the holding means, a designation means which designates an arbitrary point on the image acquired by the image acquisition means, an arrangement means (32) which arranges a measurement point on a point position on the sample corresponding to the point designated by the designation means, a measurement means (10) which measures a signal resulting from dynamic information of an object to be measured from the measurement point arranged by the arrangement means, and an analysis means (37) which analyzes measured results measured by the measurement means. <P>COPYRIGHT: (C)2005,JPO&NCIPI

Description

[0001]
TECHNICAL FIELD OF THE INVENTION
The present invention relates to a biomolecule analyzing apparatus for determining statistical properties of a plurality of specific sites in a sample, interactions between the sites, and the like.
[0002]
[Prior art]
For a confocal scanning optical microscope, see, for example, the document “Confocal Microscopy”, T.A. Wilson (ed.) Academic press (London) has commentary. Also, as a commentary mainly on biological samples, reference is made to the document “Handbook of Biological Confocal Microscopy”, J. Org. B. Pawley (ed.) Plenum Press (New York) and the like. Regarding fluorescence correlation spectroscopy, see, for example, the document “Fluorescence correlation spectroscopy”, R.A. Rigler, E .; S. There is a commentary such as Elson (eds.) Springer (Berlin).
[0003]
In the 1990s, research on single molecule detection and imaging using fluorescence has been rapidly increasing. For example, as a method for detecting a single molecule, reference M. Goodwin etc. ACC. Chem. Res. (1996), Vol. 29, p607-613, and fluorescence correlation spectroscopy (FCS). In fluorescence correlation spectroscopy, proteins and carrier particles labeled with fluorescence are suspended in a solution in the field of view of a confocal laser microscope, and fluctuations in fluorescence intensity based on Brownian motion of these particles are analyzed to obtain an autocorrelation function. Then, the number and size of the target fine particles are estimated. This technique is described, for example, in Masataka Kaneshiro, “Protein Nucleic Acid Enzyme” (1999) Vol. 44, no. 9, pp 1431-1437.
[0004]
On the other hand, in a confocal scanning optical microscope, a laser beam is used as a light source, and the laser beam is two-dimensionally scanned by a scanning mirror or the like, and fluorescence emitted from a fluorescent dye in a sample is received to obtain a two-dimensional fluorescence microscope image.
[0005]
[Problems to be solved by the invention]
In addition, several patent applications have been filed for the above-mentioned technology. In Japanese Patent Application Laid-Open No. 2001-194303, in fluorescence correlation spectroscopy, a laser beam is guided to a cylindrical lens, and thereby converted into a linear light beam. Further, the linear light beam is scanned in the vertical direction by the galvanometer mirror, and the scanning is switched to two-dimensional scanning, so that the entire imaging area of the sample is irradiated with the excitation light to excite the entire sample. Then, the fluorescence signal from the sample is detected by a two-dimensional photodetector, and the relationship between the two-dimensional position in the sample and the light intensity is obtained.
[0006]
In this method, since the whole inside of the imaging region of the sample is excited, the fluorescence signals from a plurality of locations can be extracted at the same time. However, according to this method, it is impossible to distinguish and extract the fluorescence signals emitted from a plurality of desired minute specific parts distributed three-dimensionally in the imaging region of the sample. Further, in this method, since the whole of the imaging region of the sample is excited, a fluorescence signal from the sample can be captured, but is limited to a confocal region irradiated with laser light. Therefore, when a sample such as a cell moves and moves out of the confocal region, the sample stage must be moved to an optimal position.
[0007]
Further, in JP-A-08-43939 and JP-A-2000-98245, in a scanning optical microscope, fluorescence from a multi-stained sample is separated by a grating and a prism, and the component wavelength is detected by a plurality of photodetectors. Detection is performed every time. With this method, signals for each fluorescent dye can be extracted from the entire imaging area of the sample that is excited at one time, but the fluorescence signals emitted from a plurality of selected minute specific sites in the imaging area of the sample are respectively obtained. It cannot be taken out separately.
[0008]
In Japanese Patent Application Laid-Open No. 08-068694, a plurality of optical fiber tubes are used, and a fluorescence signal is simultaneously extracted from a spatially separated portion in a sample by a plurality of photodetectors by multiplex processing to perform fluorescence correlation analysis. Even with this method, fluorescence signals can be extracted from the entire imaging region of the sample, but it is not possible to separate and extract the fluorescence signals emitted from a plurality of selected minute specific sites in the sample.
[0009]
In Japanese Patent Application Laid-Open No. 09-113448, a plurality of reaction vessels are installed, the same laser light is applied to each of them, a fluorescence signal from a sample in each reaction vessel is taken out by a confocal optical system, and a fluorescence correlation analysis is performed. Perform In this case, fluorescence signals from different sites can be obtained for a plurality of samples, and also, fluorescence signals from a plurality of sites can be obtained at the same time. However, it is not possible to perform correlation analysis by receiving a fluorescent signal while identifying a plurality of sites in the same cell. In addition, although fluorescence intensity fluctuation signals from a plurality of sites in the sample can be obtained, the signals are limited to signals within a predetermined confocal region, and cannot respond to movement of cells or the like in real time.
[0010]
Japanese Patent Application Laid-Open No. H10-206742 describes a method for obtaining dynamic characteristics of a sample by irradiating a desired position of the sample with laser light while observing the sample image in a laser scanning confocal optical microscope. . In this case, irradiation of a pulsed laser and acquisition of the dynamic characteristics of a specimen caused by the influence are disclosed, and fluctuation of the intensity of a fluorescence signal from the specimen cannot be measured.
[0011]
Living cells are continually moving, but the confocal area of the confocal microscope, which is the observation field, is very small. It may be out of focus area. In this case, in order to return the cells that have deviated from the confocal region back to the confocal region for observation, it is necessary to move the sample stage to an optimal position, which is troublesome. Alternatively, it is necessary to move the sample stage following the cell while observing the moving cell. In this case, as a method of capturing moving cells or the like, for example, an optical trap using a laser beam is usually performed.
[0012]
Regarding attempts to optically trap cells and the like with laser light, for example, in Japanese Patent Application Laid-Open No. 08-280375, a laser light is applied to a fast-moving sample such as Escherichia coli through an objective lens to stop the movement of the sample, and a micropipette is used to stop desired movement Microorganisms are sucked and captured. In Japanese Patent Application Laid-Open No. 07-104191, a YAG laser beam is applied to microparticles such as cells, viruses, and chromosomes through an objective lens to optically trap the microparticles. Posture control is performed. Also, Steven B. Smith et al. Attempt to capture latex particles to which DNA has adhered by an optical trap in a channel, extend the attached DNA, and measure its behavior and force. (Steven B. Smith etc., Science (1996) Vol. 271, 795-)
Further, Rigler et al. Attached DNA labeled with a fluorescent dye to each base to microparticles, optically trapped the DNA with laser light in a microchannel, flowed a restriction enzyme into the microchannel, and added each of the DNAs. The base is cut one by one, and the fluorescent dye attached to each base which is cut in the confocal region focused slightly downstream in the microchannel and flows in the microchannel is excited. Attempts have been made to determine the DNA base sequence by sequentially detecting the emitted fluorescence. (Bioimaging Vol. 5 (1997) p. 139-152)
An object of the present invention is to provide a biomolecule analyzing apparatus for performing multipoint fluorescence correlation analysis while fixing an observation target by an optical trap.
[0013]
[Means for Solving the Problems]
In order to solve the problem and achieve the object, the biomolecule analyzing apparatus of the present invention is configured as follows.
[0014]
(1) A biomolecule analyzing apparatus of the present invention is a biomolecule analyzing apparatus for analyzing a dynamic behavior of a biomolecule, and a holding unit for optically holding the biomolecule in a measurable state; Image acquisition means for acquiring an image corresponding to at least one observation region of the biological sample containing the biomolecules held in, and designation means for designating an arbitrary point on the image acquired by the image acquisition means Arranging means for arranging a measurement point at a point position on the sample corresponding to the point designated by the designation means, and measuring a signal derived from dynamic information of the substance to be measured from the measurement point arranged by the arranging means Measurement means for performing the measurement, and analysis means for analyzing a result measured by the measurement means.
[0015]
(2) The biomolecule analyzing apparatus according to the present invention is the apparatus according to (1), wherein the image obtained by the image obtaining means is an image including a two-dimensional or three-dimensional area, and A point is arranged at an arbitrary position in three dimensions.
[0016]
(3) The biomolecule analyzing apparatus according to the present invention is the apparatus according to the above (1), and the image acquiring means comprises a confocal scanning optical microscope using laser light.
[0017]
(4) The biomolecule analyzing apparatus according to the present invention is the apparatus according to the above (1), and the holding means holds the biomolecule by laser light.
[0018]
BEST MODE FOR CARRYING OUT THE INVENTION
(First Embodiment)
FIG. 1 is a diagram showing a basic configuration of a biomolecule analyzing apparatus according to the first embodiment of the present invention. FIG. 2 is a block diagram of a fluorescence image acquisition and correlation analysis apparatus of a biomolecule provided with an optical trap mechanism according to the present invention.
[0019]
In the present invention, based on a confocal scanning optical microscope, a sample (or a sample is attached to a carrier) such as a biological cell fluorescently labeled at a specific site in a field of view is irradiated with laser light, and a sample image is formed. While observing and recording, a correlation analysis is performed on the intensity fluctuations of the fluorescence from a plurality of specific sites in the sample, and the statistical properties of the sites and the interaction between the sites are determined. In addition, a mechanism for optically trapping a sample (or a carrier) such as a biological cell with a laser beam and holding the sample so as not to be displaced from the confocal observation region is provided. As a sample, for example, a fluorescent dye is attached to a part of the protein to be labeled, and the interaction at the time of binding to the target protein and the kinetics after the binding are analyzed. In addition, the DNA is labeled with a fluorescent dye, and the interaction of the fluorescent dye with the protein and the analysis of DNA hybridization are performed.
[0020]
In FIG. 1, laser light is two-dimensionally scanned (XY-axis scanning) using a pair of XY scanners (X-axis scanning scanner and Y-axis scanning scanner) whose rotation directions are orthogonal to each other, and the fluorescence from the sample and the fluorescence The intensity fluctuation is received by a photodetector (light receiver). Here, a galvano scanner is used as the XY scanner. The laser beam is configured to expand the beam diameter and form collimated light by a combination of a pair of lenses and a pinhole, and to guide the beam to a pair of galvano scanners.
[0021]
As the laser light, for example, an argon laser having a wavelength of 488 nm or a He.Ne laser having a wavelength of 633 nm is used. The fluorescent dye for labeling a desired portion in the cell is rhodamine green (RhG) when an argon laser having a wavelength of 488 nm is used as excitation light, and sifive (Cy5) when using a He / Ne laser having a wavelength of 633 nm as excitation light. ) May be used.
[0022]
The collimated laser light is focused on a desired cell site in the sample S by the objective lens 5. The laser beam focused on a desired portion in the sample S is two-dimensionally scanned by a galvano scanner, and an image is formed by capturing a fluorescent signal of a fluorescent dye in the sample S. Here, the galvano scanner is stopped at the required position, the intensity fluctuation of the fluorescence from the fluorescent dye molecule is received by the photodetector 10 as a time-series signal, and a correlation analysis operation is performed by a computer, and the autocorrelation function of the fluorescence intensity fluctuation is calculated. can get.
[0023]
In order to obtain a two-dimensional microscope image of the sample S, two-dimensional scanning of the laser light source 1 is required. For example, two galvanometer mirrors are arranged so that the directions of rotation are orthogonal to each other, and an X-axis scanning scanner 61 and a Y-axis scanning scanner 62 are used to scan the XY axes with laser light. The cell is irradiated with laser light by scanning the XY axes, reflected light and fluorescence are received by the photodetector 10, and an image signal is generated by an image processing device 35 via a signal processing device 34 shown in FIG. A two-dimensional image of the sample S is generated on the monitor 36.
[0024]
The scanning angle of the XY scanner is detected by the XY scanner angle detection device 33, and the detection signal is guided to the computer 37, combined with the image signal output from the image processing device 35, and the microscope of the sample S on the screen of the TV monitor 36. The position of the image is matched. On the other hand, the same optical system is used to excite fluorescent dye molecules by laser light. Thus, the sample S is irradiated with the laser beam, the fluorescence signal from the fluorescent molecule is received by the photodetector 10, and the computer 37 performs the fluorescence correlation analysis.
[0025]
Designation of a scanning point on a two-dimensional microscope image is performed as follows. The observer finds light emission from the fluorescent molecule at a desired point while viewing the observation image on the TV monitor 36, and stops the XY scanner 31 from scanning at this point using an operating unit (designating means) (not shown) such that the XY scanner 31 stops scanning. Adjust the scanner drive 32. As a result, a measurement point is arranged at an arbitrary position on the three-dimensional surface of the sample S. The scanning angle of the XY scanner 31 at this time is accurately detected by the XY scanner angle detecting device 33.
[0026]
Since the scanning optical system, the excitation optical system, and the detection optical system have different wavelengths, the dichroic mirror 7 separates the optical systems from each other. Note that, in order to perform XY scanning with laser light, an acousto-optic modulator (AOM), a polygon mirror, or the like may be used instead of a galvano scanner.
[0027]
The focus movement of the laser light can be performed by a method such as moving the objective lens 5 in the optical axis direction. Thus, a three-dimensional image of the sample S can be generated on the TV monitor 36. In addition, the irradiation position of the laser beam can be three-dimensionally moved to a desired position inside or outside the cell simultaneously with the image acquisition.
[0028]
On the other hand, an optical trapping optical system using laser light uses a YAG laser having a wavelength of 1,064 nm as the laser light source 20 for optical trapping, and expands the beam diameter with an optical system including a plurality of lenses and pinholes, and then converts the beam into parallel light. After being guided to the dichroic mirror 71 and joined to the measurement optical system, it is guided to the objective lens 5 to be focused in the confocal region, and captures desired fine particles and the like in the sample S. At this time, cells, chromosomes, fine particles, and the like are fixed in the confocal region by the trap light, and the measurement can be performed without departing from the confocal region due to Brownian motion or the like.
[0029]
In FIG. 2, after a fluorescence signal from the sample S obtained through the objective lens 5 is received by the photodetector 10, image processing such as contrast enhancement and contour enhancement is performed together with image formation by the image processing device 35. It is guided to a computer 37 and a two-dimensional image is obtained on a TV monitor 36. The sample S is irradiated with the laser light from the laser light source 1, and the intensity fluctuation of the fluorescence from the fluorescent molecules is received by the photodetector 10 as a time-series signal and guided to the computer 37, where the autocorrelation function is obtained. Can be
[0030]
When the intensity of the intensity fluctuation of the fluorescence from the fluorescent molecule is relatively large, an analog time-series signal is obtained. When the intensity of the intensity fluctuation of the fluorescence from the fluorescent molecule is very small, the intensity fluctuation of the intensity of the fluorescence from the fluorescent molecule becomes a photocurrent pulse. From the obtained autocorrelation function, statistical properties such as the speed of the translational diffusion motion of the fluorescent molecule are obtained. Further, a laser light source 20 for an optical trap is provided in another optical path, merges with the optical system for measurement, and is introduced into the objective lens 5.
[0031]
(Second embodiment)
FIG. 3 is a diagram showing a basic configuration of a biomolecule analyzing device according to the second embodiment of the present invention. 3, the same parts as those in FIG. 1 are denoted by the same reference numerals. In FIG. 3, the basic configuration of the optical system is the same as that of the first embodiment, and two photodetectors 10a and 10b are arranged so as to face each other. That is, cross-correlation analysis of fluorescence intensity fluctuations is performed by arranging two light receivers at an arbitrary angle around the confocal region with respect to the confocal region of one optical system. Can be. The arrangement of the two photodetectors 10a and 10b is not limited to this. For example, the two photodetectors 10a and 10b may be arranged in directions in which the optical paths are substantially perpendicular to each other.
[0032]
(Third embodiment)
FIG. 4 is a diagram showing a basic configuration of a biomolecule analyzing apparatus according to the third embodiment of the present invention. 4, the same parts as those in FIG. 1 are denoted by the same reference numerals. FIG. 4 includes an optical system that performs image acquisition and fluorescence correlation analysis of two cells and the like in the sample S. In the third embodiment, the excitation laser light is focused on two spatially different points in the sample S by the two excitation optical systems. Thereby, fluorescence images at two different places in the sample S can be obtained, and fluorescence correlation analysis can be performed.
[0033]
As the laser light as the excitation light for the fluorescent dye molecules, for example, an argon laser having a wavelength of 488 nm and a He.Ne laser having a wavelength of 633 nm are used. Rhodamine green (RhG) and sifive (Cy5) are used as fluorescent dyes for labeling a desired portion in a cell. Rhodamine green (RhG) is excited with an argon laser at a wavelength of 488 nm. Sifive (Cy5) is excited by a 633 nm He.Ne laser.
[0034]
The fluorescence from the fluorescent dye molecule at the desired location passes through the same optical path as the excitation light, and is separated from the incident optical path by the dichroic mirrors 7 and 7b adjusted according to the emission wavelengths of the respective fluorescent dyes. , 10b. As the photodetectors 10a and 10b, photomultiplier tubes or avalanche photodiodes are used. The time-series signal of the photocurrent of the light intensity of the fluorescent light received by the photomultiplier tube is guided to the signal processing device 34 shown in FIG. 2, converted into a voltage signal, waveform-shaped, and binarized on-off. The signal is converted into a pulse and guided to the computer 37 as a digital signal. A correlation operation is performed on the binarized pulse signal input to the computer 37 to obtain an autocorrelation function. Further, from the obtained autocorrelation function, the translational diffusion speed of the fluorescent molecules, the change in the number density of the fluorescent dye molecules existing in the measurement region, and the like are obtained.
[0035]
FIG. 5 is a block diagram of a biomolecule fluorescence image acquisition and correlation analysis apparatus provided with an optical trap mechanism according to the third embodiment of the present invention. 5, the same parts as those in FIG. 4 are denoted by the same reference numerals. FIG. 5 includes an optical system that performs image acquisition and fluorescence correlation analysis of two cells and the like in the sample S. The measuring optical system has two optical paths, an autocorrelation function can be obtained from two spatially discrete points in the sample S, and image acquisition and fluorescence correlation analysis of two cells in the sample S can be performed. .
[0036]
In the third embodiment, two-dimensional scanning is performed by two scanning optical systems to obtain a two-dimensional image of a sample such as a cell, and to reduce the intensity fluctuation of the fluorescence due to the fluorescent dye present in the cell or the like in the sample. Perform fluorescence correlation analysis. By adding a lens for adjusting the focal length to the two scanning optical systems and moving the focus position on the optical axis, a three-dimensional image of the sample can be obtained. Alternatively, a three-dimensional image of the sample can be similarly obtained by adding a lens to each scanning optical system and moving the focal length of the lens.
[0037]
(Fourth embodiment)
FIG. 6 is a block diagram of a fluorescence image acquisition and correlation analysis apparatus for biomolecules provided with an optical trap mechanism according to a fourth embodiment of the present invention. 6, the same parts as those in FIG. 2 are denoted by the same reference numerals. In FIG. 6, three optical paths are provided independently of each other, and three laser light sources having different wavelengths are used.
[0038]
As the laser light, for example, an argon laser having a wavelength of 488 nm, a He.Ne laser having a wavelength of 543 nm, and a He.Ne laser having a wavelength of 633 nm are used. Examples of the fluorescent dye include rhodamine green (RhG) excited by an argon laser having a wavelength of 488 nm, tetramethylrhodamine (TMR) excited by a He / Ne laser having a wavelength of 543 nm, and cyfive excited by a He / Ne laser having a wavelength of 633 nm. And so on. As a result, it is possible to acquire microscope images at three different places in the sample and perform fluorescence correlation analysis of cells and molecules.
[0039]
(Fifth embodiment)
7 and 8 are block diagrams of a fluorescence image acquisition and correlation analysis apparatus for biomolecules provided with an optical trap mechanism according to a fifth embodiment of the present invention. 7, the same parts as those in FIG. 5 are denoted by the same reference numerals. 8, the same parts as those in FIG. 6 are denoted by the same reference numerals.
[0040]
In the fifth embodiment, the optical system of the optical trap forms an optical path different from the optical system for measurement, and the laser light from the laser light source 20 for optical trap faces the optical system for measurement from the opposite surface of the sample S. be introduced. Thereby, it is possible to perform image acquisition and fluorescence correlation analysis of two cells and the like in the sample.
[0041]
(Sixth embodiment)
FIG. 9 is a block diagram of a biomolecule fluorescence image acquisition and correlation analysis apparatus including an optical trap mechanism according to the sixth embodiment of the present invention. FIG. 9 includes an optical system that performs image acquisition and fluorescence correlation analysis of four discrete cells in a sample.
[0042]
In the sixth embodiment, two independent two-dimensional scans are performed by four independent scanning optical systems to obtain a two-dimensional image of a sample such as a cell, which is present in four different cells in the sample. Fluorescence correlation analysis is performed from the fluctuation of the intensity of the fluorescence by the fluorescent dye. Further, by adding a lens for adjusting the focal length to the four scanning optical systems and moving the focus position on the optical axis, it is possible to obtain three-dimensional images at four different places on the sample.
[0043]
(Seventh embodiment)
FIG. 10 is a block diagram of a fluorescence image acquisition and correlation analysis apparatus for biomolecules provided with an optical trap mechanism according to a seventh embodiment of the present invention. In FIG. 10, the same parts as those in FIG. 9 are denoted by the same reference numerals.
[0044]
In the seventh embodiment, the optical system of the optical trap forms an optical path different from that of the optical system for measurement, and the laser light from the laser light source 20 for optical trap faces the optical system for measurement from the opposite surface of the sample S. Is introduced. Thus, an optical path different from the optical system that performs image acquisition and fluorescence correlation analysis of images of cells at four locations in the sample is formed, and light trapping can be performed from the opposite surface of the sample.
[0045]
The biomolecule analyzer of the embodiment described above is based on a confocal scanning optical microscope, acquires a fluorescence microscope image of a desired cell or molecule while capturing fine particles such as a living cell, virus, or chromosome, and observes it. , And record, and perform the fluorescence correlation analysis. In addition, the optical trap captures the cell, or a part of the cell, chromosome, fine particles, or the whole cell, and prevents the observation sample from leaving the observation field. Further, the region to be observed of the sample such as a cell is fixed so as to stay at a desired position in the observation visual field. Alternatively, a region to be observed of a sample such as a cell is moved to a desired position in an observation visual field and fixed.
[0046]
This prevents the cells from disappearing from the confocal observation region due to Brownian motion or the like during the observation, and eliminates the need to move the stage. In addition, it is possible to acquire fluorescence microscope images of a plurality of discretely existing portions in the sample while holding the sample, and perform a fluorescence correlation analysis of a plurality of discretely desired cells and molecules.
[0047]
As described above, while the sample is held in the confocal observation region by performing the optical trap with the laser light, the laser light can be simultaneously focused on a plurality of locations existing in three-dimensionally different portions, and the discrete Fluorescence correlation analysis of a plurality of desired cells or molecules can be performed.
[0048]
Note that the present invention is not limited to the above embodiments, and can be appropriately modified and implemented without changing the gist. For example, by further increasing the number of measurement optical systems in parallel to perform fluorescence correlation analysis of desired cells or molecules, fluorescence microscopic images of a plurality of discrete sites may be acquired, displayed, and recorded. Further, in the above-described embodiment, measurement using fluorescence correlation spectroscopy has been described as an example, but the present invention is not limited to this. That is, the present invention is applicable to any micro-optical measurement for measuring various optical properties (polarization, scattering, electrochemiluminescence, resonance energy transfer, plasmon resonance, etc.) limited to a specific site or a specific region of a sample to be measured. Applicable. Further, the image to be acquired is not limited to a temporary two-dimensional or three-dimensional image, and may be a still image or a video image over a plurality of hours. Also, the sample holding means can be cells or other components capable of appropriate optical holding (containers, solution components, temperature controllers, optical element materials, etc.).
[0049]
【The invention's effect】
ADVANTAGE OF THE INVENTION According to this invention, the biomolecule analysis apparatus which performs a fluorescence correlation analysis of multiple points while fixing an observation object with an optical trap can be provided.
[Brief description of the drawings]
FIG. 1 is a diagram showing a basic configuration of a biomolecule analyzing apparatus according to a first embodiment of the present invention.
FIG. 2 is a block diagram of a biomolecule fluorescence image acquisition and correlation analysis apparatus provided with the optical trap mechanism according to the first embodiment of the present invention.
FIG. 3 is a diagram showing a basic configuration of a biomolecule analyzing apparatus according to a second embodiment of the present invention.
FIG. 4 is a diagram showing a basic configuration of a biomolecule analyzing apparatus according to a third embodiment of the present invention.
FIG. 5 is a block diagram of a biomolecule fluorescence image acquisition and correlation analysis apparatus including an optical trap mechanism according to a third embodiment of the present invention.
FIG. 6 is a block diagram of a biomolecule fluorescence image acquisition and correlation analysis apparatus including an optical trap mechanism according to a fourth embodiment of the present invention.
FIG. 7 is a block diagram of a biomolecule fluorescence image acquisition and correlation analysis apparatus including an optical trap mechanism according to a fifth embodiment of the present invention.
FIG. 8 is a block diagram of a fluorescence image acquisition and correlation analysis apparatus for biomolecules provided with an optical trap mechanism according to a fifth embodiment of the present invention.
FIG. 9 is a block diagram of a biomolecule fluorescence image acquisition and correlation analysis apparatus including an optical trap mechanism according to a sixth embodiment of the present invention.
FIG. 10 is a block diagram of a biomolecule fluorescence image acquisition and correlation analysis apparatus including an optical trap mechanism according to a seventh embodiment of the present invention.
[Explanation of symbols]
1, 1a, 1b, 1c laser light source 2 first lens 3 pinhole for illumination light 4 second lens 5, objective lens 61, 61a, 61b X-axis scanning scanner 62, 62a, 62b Y-axis Scanning scanner 7 Dichroic mirror 8, 8a, 8b Light receiving pinhole 9, 9a, 9b Light receiving lens 10, 10a, 10b, 10c Photodetector 11 Dichroic mirror 20 Laser light source for optical trap 21, 22 ... Lens 23 ... Pinhole 32 ... XY scanner driving device 33 ... XY scanner angle detecting device 34 ... Signal processing device 35 ... Image processing device 36 ... TV monitor 37 ... Computer

Claims (4)

A biomolecule analyzer for analyzing dynamic behavior of biomolecules,
Holding means for optically holding the biomolecules in a measurable state,
Image acquisition means for acquiring an image corresponding to at least one observation region of the biological sample containing the biological molecule held by the holding means,
Specifying means for specifying an arbitrary point on the image obtained by the image obtaining means;
Arranging means for arranging a measurement point at a point position on the sample corresponding to the point specified by the specifying means;
Measuring means for measuring a signal derived from the dynamic information of the substance to be measured from the measuring points arranged by the arranging means,
Analysis means for analyzing the result measured by the measurement means;
A biomolecule analysis device comprising: The image obtained by the image obtaining means is an image including a two-dimensional or three-dimensional region,
2. The biomolecule analyzing apparatus according to claim 1, wherein the arranging unit arranges the measurement points at an arbitrary position in three dimensions. 2. The biomolecule analyzing apparatus according to claim 1, wherein the image acquisition unit comprises a confocal scanning optical microscope using laser light. The biomolecule analyzing apparatus according to claim 1, wherein the holding unit holds the biomolecule using laser light.

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