JP6617978B2 - Optical imaging device - Google Patents

Description

  The present invention irradiates a biological sample with light, and two-dimensionally detects various light such as transmitted light, reflected light, or fluorescence obtained from the sample, thereby optical properties of the sample. In particular, the present invention relates to an optical imaging apparatus that acquires an image showing characteristics and features.

  When excitation light emitted from a light source such as a halogen lamp or a semiconductor laser is irradiated to a small animal such as a mouse or a rat to which a fluorescently labeled probe is administered, the fluorescently labeled probe is collected from a site such as an organ where the probe is integrated. Fluorescence is emitted. By detecting this fluorescence with a highly sensitive camera and imaging its intensity distribution, the distribution and behavior of the molecular species of interest are non-invasive to a small animal in a living state (in vivo). You can investigate. As an apparatus for performing such measurement, for example, optical imaging apparatuses described in Patent Document 1, Non-Patent Documents 1 and 2, and the like are known.

  In a conventional general optical imaging apparatus, a biological sample (such as a small animal) placed on a stage whose upper surface is substantially horizontal is irradiated with illumination light from above, and excited and emitted by the reflected light or illumination light. The detected fluorescence is detected by a camera disposed above the biological sample, or the biological sample placed on the stage is irradiated with illumination light from below, and the fluorescence excited and emitted by the illumination light is emitted. A configuration is adopted in which detection is performed by a camera disposed above the biological sample. In order to avoid the influence of outside light, the entire optical system from the light source to the camera is usually housed in a casing that ensures high light shielding properties.

  However, any of the conventional optical imaging apparatuses adopting the above-described optical system configuration includes various optical components for detecting light from the sample, such as a condensing lens system, an optical filter, and a camera. Since they are arranged side by side along an axis extending in the vertical direction, the overall height of the apparatus tends to increase. Also, generally, just increasing the overall height of the device has a risk of falling stability and falling down, so the width and depth of the device must be increased to match the height. For this reason, conventional optical imaging devices are generally large and heavy, and it is necessary to secure a wide installation place, it is necessary to increase the weight resistance of the floor surface of the installation place, and it is inconvenient to move. was there.

JP 2009-257777 A

Yajima and 13 others, “Development of Clairvivo OPT for in vivo fluorescence imaging system for small animals”, Shimadzu Editorial Department, Shimazu Review, Vol. 66, No. 1, Issue 2, September 30, 2009, p.21-27 "IVIS Imaging System (Caliper)", Sumisho Pharma International Co., Ltd., [online], [searched on November 22, 2012], Internet <URL: http://www.summitpharma.co.jp/english/service /products/xenogen/index.html#1>

  SUMMARY OF THE INVENTION The present invention has been made to solve the above-mentioned problems, and its main object is to provide an optical imaging apparatus that is easy to handle and realizes a small size and light weight by devising a configuration of an optical system. It is.

The present invention was made in order to solve the above-irradiated with excitation light to the biological sample, the optical imaging device for two-dimensional imaging by detecting fluorescence obtained from the sample with respect thereto,
a) a sample stage for placing a biological sample substantially horizontally, and a sample stage provided with a light transmitting part through which light is transmitted ;
b) saw including a plurality of emission units for emitting excitation light irradiated on a living body placed on the sample on the sample stage, spatial excitation light beam exit portion of said plurality of respectively, using optical refractive element A lighting unit that emits while diverging into ,
an imaging unit for acquiring an image by the fluorescence emitted from the biological sample to the excitation light by c) the illumination unit,
d) The fluorescence excitation light passed through the light transmitting portion is released from the biological sample downwardly guides the excitation light to be irradiated from below the biological sample through the sample stage of the light transmission portion by the illumination unit the order guided to the imaging unit, and the bending and the excitation light and the fluorescence, both in space, light guide optical system including a common reflective optical unit, which is arranged below the sample stage,
It is characterized in that to obtain Bei the.

In the optical imaging apparatus according to the present invention, the excitation light emitted from each emitting part of the illuminating part hits the reflecting optical part of the light guide optical system, the optical path thereof is bent, and the biological sample placed on the sample stage is viewed from below. Irradiated. Some are excited by the excitation light of that in the sample of the fluorescence emitted downward strikes the reflective optical unit proceeds to the excitation light in the opposite direction, it reaches the image pickup unit optical path is bent. Since this is common in the reflecting optical unit and the excitation light and fluorescence to be bent together to place the optical components and elements included in the optical parts and elements and imaging unit included in the lighting unit arranged in the vertical direction There is no need, for example, they may be arranged in the horizontal direction. As a result, the overall height of the apparatus can be suppressed.
In addition, according to the present invention, since the reflection optical unit is not disposed in the upper space of the sample table, the upper part of the sample table can be easily opened and closed. With such a structure, the biological sample is placed on the sample table from above. Can be placed. In addition, a fluorescent image or a visible light image on the abdomen side of the biological sample can be acquired in a state where the biological sample maintains a natural posture.

In the optical imaging apparatus according to the present invention, each emitting unit emits an excitation light beam while spatially diverging it using an optical refractive element such as a lens array. In general, a laser beam emitted from a light source such as a semiconductor laser has a high energy density but a small spot diameter. On the other hand, in the present invention, by diverging the laser light beam with an optical refracting element such as a lens array, the light beam can be expanded and a certain wide range can be illuminated with a substantially uniform light intensity.

In the optical imaging device according to the present invention, preferably, the plurality of emission units are more disposed to surround the optical axis of the fluorescence towards the said imaging unit from said reflecting optical unit, the optical from the biological sample in the optical axis substantially coaxial fluorescence towards the reflecting portion from the reflected light faculty may be configured to excitation light to the biological sample is illuminated. More preferably, it may be arranged Te than at substantially the same rotational angle spacing surrounds the optical axis of the fluorescence towards the imaging unit a plurality of emission units from the reflective optical unit.

According to this arrangement, even if no located illumination unit on the optical axis of the fluorescence towards the imaging unit from the reflective optical unit, excitation intensity of substantially uniform with respect to a living body placed on the sample on a sample stage Light can be irradiated. Thereby, in the case so as to obtain a fluorescence intensity distribution image of the fluorescent substance derived incorporated inside the raw body samples, there is no excitation light intensity distribution dependent, it is possible to obtain accurate fluorescence intensity distribution image .

Each of the plurality of emission units may have a light source, but preferably, each of the measurement lights emitted from one light source such as a high-intensity semiconductor laser is divided into a plurality of parts by, for example, an optical fiber. It is good to set it as the structure which transmits to an injection | emission part and inject | emits toward the space from an injection | emission part. Thereby, the wavelength of the excitation light emitted from each emission part can be made the same, and the cost of the apparatus can be suppressed.

In the optical imaging device according to the present invention may further comprise an optical dividing unit which divides the fluorescence from the biological sample to a plurality of wavelength regions of light, the imaging unit includes a plurality of divided by the optical dividing portion It is good also as a structure containing the some imaging part which each acquires the image by the light of a wavelength range.

For example, when fluorescence from the biological sample contains a reflected light fluorescence and visible wavelength region of the infrared or near infrared wavelength region, the above configuration, to obtain a fluorescence image and a visible light image of the biological sample simultaneously Can do. Further, since the fluorescence also includes reflected light of the excitation light may be captured respectively by separating the excitation light wavelength range and the fluorescence wavelength range. Thereby, since the two-dimensional intensity distribution of the excitation light is known, the uniformity of the excitation light irradiation intensity can be evaluated, or the adjustment for making the excitation light irradiation intensity uniform can be performed.

Moreover, an optical imaging apparatus according to the present invention includes:
A movement in which the photometric unit including the illumination unit, the light guide optical system, and the imaging unit and the sample stage are relatively moved in a one-dimensional or two-dimensional manner parallel to the mounting surface of the sample stage. A mechanism part;
An operation unit that is operated by a measurer so that the photometric unit and the sample stage are in a desired positional relationship by the moving mechanism unit;
And a configuration in which a measurer performs an operation using the operation unit so that a two-dimensional image by light obtained from an arbitrary part of a biological sample placed on the sample stage can be acquired. it can.

  Here, the moving mechanism unit does not include a driving source such as a motor, and the sample table or the photometric unit may be mechanically moved according to the amount of operation of the operating unit by the measurer, but preferably It is better to move the sample stage or the photometry unit by electrical control.

  That is, in the optical imaging apparatus according to the present invention, the moving mechanism unit may include a drive source, and may further include a control unit that controls the drive of the drive source according to the operation of the operation unit by the measurer.

  According to these configurations, even when the sample is considerably larger than the range that can be photographed by the photometry unit, the measurer operates the operation unit to appropriately select the site that he or she wants to observe or measure. Or it can be measured. Of course, it is also possible to obtain a fluorescent image over a wide range by repeating observation or measurement while operating the operation unit.

Further, an optical imaging apparatus according to the present invention includes:
A movement in which the photometric unit including the illumination unit, the light guide optical system, and the imaging unit and the sample stage are relatively moved in a one-dimensional or two-dimensional manner parallel to the mounting surface of the sample stage. A mechanism part;
An imaging control unit that acquires at least one of the photometry unit or the sample stage in a one-dimensional or two-dimensional manner by the moving mechanism unit, and acquires an image of light emitted from the biological sample a plurality of times by the imaging unit;
It can also be set as the structure further provided.

According to this configuration, even if the sample is considerably larger than the range that can be photographed by the photometry unit, the fluorescent image in a wider range than the photographable range can be obtained without the troublesome operation and work by the measurer himself / herself. Etc. can be acquired automatically.
In the above-described configuration, in particular, an image forming unit that reproduces a two-dimensional image in a wider range than a range obtained by one shooting by the imaging unit by combining a plurality of images acquired under the control of the shooting control unit If it is set as the structure further equipped with, it is not the picked-up image with respect to a different site | part, but can obtain one image which connected them.

According to the biological observation apparatus according to the present invention, excitation light is irradiated to the biological sample, since both of the fluorescent light obtained from a biological sample is bent by a common reflective optical unit, from the true bottom of its relative biological sample , that is, the excitation light to the biological sample is irradiated along an axis extending in the vertical direction, even in a configuration in which a biological sample or fluorescent light is removed and along the same axis, Ya various optical components included in illumination unit and the imaging unit The optical elements can be arranged side by side in the horizontal direction instead of the vertical direction. Thereby, the height of the apparatus can be suppressed, and the entire apparatus can be made compact. Moreover, since stability is also improved by suppressing the overall height of the apparatus, it is possible to avoid an increase in weight more than necessary, and it is possible to reduce the weight of the apparatus.

Also the optical imaging device according to the present invention, irradiated with excitation light from below, since a configuration for observing the fluorescence downward, when the biological sample is a small animal such as mice and rats, so-called supine The abdominal side can be observed in a natural posture with the abdomen facing down instead of an unnatural posture. Thereby, it is possible to avoid placing an excessive burden on the small animal, thereby increasing the reliability of the measurement.

1 is an external perspective view of an optical imaging apparatus according to an embodiment of the present invention in a state in which a sample mounting upper lid is opened. FIG. The schematic side view in the state where the exterior case of the optical imaging device of this example was removed. 1 is a configuration diagram of an optical system in an optical imaging apparatus according to the present embodiment. The figure which shows arrangement | positioning of the emission part and visible light emission part in the optical imaging apparatus of a present Example. The block diagram of the principal part of the optical imaging apparatus which is another Example of this invention. The block diagram of the principal part of the optical imaging apparatus which is another Example of this invention.

Hereinafter, an optical imaging apparatus according to an embodiment of the present invention will be described with reference to the accompanying drawings.
FIG. 1 is an external perspective view of an optical imaging apparatus according to an embodiment of the present invention in a state in which a sample mounting upper lid is opened, and FIG. FIG. 3 is an optical system configuration diagram of the optical imaging apparatus of the present embodiment. That is, FIG. 2 is a diagram specifically showing a spatial arrangement of each component and each element of the optical system in the optical imaging apparatus of the present embodiment, and FIG. 3 is a diagram showing a functional configuration of the optical system. is there.

  As shown in FIG. 1, the optical imaging apparatus of the present embodiment has a substantially box shape that is longer in the depth direction than the width and height, and can be swung upward at the upper front part of the outer casing 20. A sample mounting upper lid 21 is provided. When the sample mounting upper lid 21 is opened, the sample table 5 whose upper surface is substantially horizontal is exposed, and a biological sample 7 such as a mouse or a rat to be measured can be mounted on the sample table 5. . When the sample placement upper lid 21 is opened, there are no obstacles above the sample stage 5, so that the measurer can place or take out the biological sample 7 very easily.

As shown in FIG. 2, the sample stage 5 is large and rectangular metallic support base 5 which opening window 6 is formed of b in the center, a transparent plate such as glass mounted on the said support base 5 b 5 a . By the transparent plate 5a and the opening window 6, the lower surface of the biological sample 7, that is, the abdominal surface of the biological sample 7 can be seen through from below in a normal placement state. In the lower space of the sample stage 5, a reflecting mirror 4 corresponding to the reflecting optical part in the present invention and four emitting units 3 each corresponding to the emitting part in the present invention are arranged. In addition, an objective lens 10, a spectroscopic unit 11, a fluorescent photographing camera 12, and the like are disposed behind the space in which these are disposed (to the right in FIG. 2), and above that, the laser light source unit 1 and visible photographing. A camera 13 is disposed. Note that some optical components and elements to be described later do not appear in FIG.

With reference to FIG. 3 in addition to FIG. 2, the configuration of the optical system and the function of each optical component / element in the optical imaging apparatus of the present embodiment will be described.
Prior to the measurement, a probe labeled with a predetermined fluorescent substance is administered to the biological sample 7 such as a mouse to be measured. As shown in FIG. 1, the measurer opens the sample placement upper lid 21, places the living biological sample 7 on the sample stage 5, and closes the sample placement upper lid 21.

  When the laser light source unit 1 is driven by a measurement start instruction by a measurer, the measurement light emitted from the laser light source unit 1 (in this case, excitation light) is branched into four through the optical fiber 2 and then 4 Guided to the individual injection units 3. Each emission unit 3 includes an optical refracting element that is a lens array, and divides light so that a relatively small-diameter light emitted from the end of the optical fiber 2 is expanded to a predetermined angle and intensity unevenness in the illumination area is reduced. While ejaculating. The four emission units 3 are arranged substantially rotationally symmetrically at 90 ° rotation angle intervals so as to surround an optical axis C of an objective lens 10 to be described later.

  A schematic arrangement of the four injection units 3 (3a, 3b, 3c, 3d) and the visible light emitting unit 8 is shown in FIG. Each of the four emission units 3 (3a, 3b, 3c, 3d) is fixed by being inclined by a predetermined angle so that the optical axis of the emitted light faces inward, that is, approaches the optical axis C. As a result, the light emitted from the four ejection units 3 overlaps at a position away from the four ejection units 3 by a predetermined distance, and an illumination area B having a substantially uniform light intensity is formed.

As shown in FIG. 2, a reflecting mirror 4 fixed at an angle of 45 ° with respect to the optical axis C is disposed in front of the traveling direction of the measurement light emitted from the four emission units 3 into the space. ing. Therefore, each of the measurement lights hits the reflecting mirror 4 and is bent upward at a substantially right angle. Then, the measurement light passes through the opening window 6 and the transparent plate 5 a of the sample stage 5 and strikes the lower surface of the biological sample 7. Usually, since the biological sample 7 is placed as shown in FIGS. 1 and 2, the measurement light strikes the abdominal surface of the biological sample 7.

Since the measurement light emitted from the four injection units 3 (3a, 3b, 3c, 3d) overlaps near the upper surface of the sample stage 5, the biological sample 7 is illuminated with a substantially uniform light intensity. By this light irradiation, the fluorescent substance in the biological sample 7 is excited and emits fluorescence. Fluorescence is emitted in various directions instead of one direction. Some of fluorescence emitted downward passes through the transparent plate 5 a and the opening window 6, strikes the reflector 4 located below. This fluorescence is bent substantially perpendicularly in the direction opposite to the previous measurement light (excitation light) by the reflecting mirror 4 and guided to the objective lens 10. Then, the fluorescent light focused by the objective lens 10 is introduced into the fluorescent photographing camera 12 through the spectroscopic unit 11 and forms an image on the image sensor of the camera 12. Thereby, in the fluorescence imaging camera 12, a two-dimensional image showing the intensity distribution of the fluorescence from the biological sample 7 is created, and the image is displayed and recorded on a monitor (not shown).

  The light reaching the objective lens 10 includes not only the wavelength component of fluorescence derived from the biological sample 7 but also the wavelength component of excitation light. If the image sensor mounted on the fluorescence imaging camera 12 is not sensitive to the wavelength component of the excitation light, there is no problem even if the excitation light is incident, but if it is sensitive to the wavelength component of the excitation light. It is necessary to remove the wavelength component of the excitation light before the light enters the camera 12. Therefore, in that case, the wavelength component of the excitation light is removed on the optical path until light enters the fluorescence imaging camera 12 (between the spectroscopic unit 11 and the fluorescence imaging camera 12 in the example of FIG. 3). An optical filter 14 having wavelength characteristics may be disposed, and unnecessary wavelength components of the excitation light may be removed by the optical filter 14.

  A visible light emitting unit 8 such as a visible light LED disposed between the four emission units 3 emits visible light in a predetermined wavelength range in substantially the same direction as the measurement light. The visible illumination light emitted from the visible light emitting portions 8 is bent upward by the reflecting mirror 4 and hits the lower surface of the biological sample 7 in the same manner as the measurement light. Then, the reflected light with respect to the visible illumination light travels downward from the biological sample 7 in the same manner as the above-described part of fluorescence, and is bent substantially laterally by the reflecting mirror 4 and guided to the objective lens 10. Visible reflected light that has passed through the objective lens 10 is introduced into the spectroscopic unit 11 like fluorescence, but the spectroscopic unit 11 selectively reflects only the wavelength component of visible light and transmits the near infrared to infrared wavelength components. A two-color mirror is provided. Therefore, the introduced visible reflected light is bent upward by the two-color mirror and guided to the visible photographing camera 13. As a result, a visible light image of the biological sample 7 is obtained by the visible imaging camera 13.

  Thus, in the optical imaging apparatus of the present embodiment, a visible light image and a fluorescence image on the lower surface of the biological sample 7, that is, the abdomen side, can be acquired simultaneously. Of course, obtaining a visible light image is not essential for performing fluorescence imaging.

  In the optical imaging apparatus of the present embodiment, by using the common reflector 4, the measurement light and visible illumination light irradiated on the biological sample 7 and the fluorescence and reflected light obtained from the sample 7 are both bent by approximately 90 °, Detection of the illumination optical system such as the emission unit 3, the objective lens 10, the spectroscopic unit 11, the fluorescence imaging camera 12, etc. while irradiating the biological sample 7 with measurement light and taking out fluorescence from the sample 7 in the vertical direction. Optical components and elements of the optical system are arranged side by side in the horizontal direction. As a result, the overall height of the apparatus can be suppressed as compared with the conventional apparatus, and the apparatus can be reduced in size and weight.

  In addition, the lower surface of the biological sample 7 placed on the sample stage 5 is irradiated with measurement light or visible illumination light, and fluorescence or reflected light emitted from the lower surface is detected. Side observation can be made.

  In the configuration of the above-described embodiment, the reflecting mirror 4 is a plane mirror. However, by providing the reflecting mirror 4 with power, that is, by using a concave mirror or a convex mirror, the width of the measurement field or the illumination field (that is, the illumination range) Or imaging range) can be performed. Further, if the reflecting mirror 4 is configured to be switchable or exchangeable with a plane mirror, a concave mirror, and a convex mirror, a more accurate image can be acquired according to the photographing purpose.

  Moreover, in the said Example, although the fluorescence image and visible light image of the biological sample 7 can be acquired, the wavelength component of excitation light is isolate | separated and taken out in the spectroscopic unit 11, and this excitation light is further sent to another camera. By introducing and imaging, a two-dimensional distribution of excitation light intensity can be obtained. Such a two-dimensional distribution of the excitation light intensity can be used, for example, by the operator to visually check whether there is any uneven illumination of the excitation light, or automatically irradiated so that the uneven illumination of the excitation light is reduced. Can be used to adjust.

  In addition, some fluorescent substances emit different wavelengths of fluorescence for one excitation light. Labeling with such fluorescent substances is performed, and multiple wavelengths of fluorescence are separated and introduced into separate cameras. It is good also as a structure imaged. Thereby, the two-dimensional distribution image for every fluorescence of a different wavelength can be obtained.

  Next, an optical imaging apparatus according to another embodiment of the present invention will be described with reference to FIG. FIG. 5 is a block diagram of the main part of the optical imaging apparatus of this embodiment. In the optical imaging apparatus of the above embodiment, a fluorescence intensity distribution image of a predetermined range of the biological sample 7 placed on the sample stage 5 is obtained, and the photographing range is determined. For this reason, for example, when the size of the biological sample is large and the imaging range of the sample is wider than the imaging range determined by the apparatus, multiple imagings are performed while changing the mounting position of the biological sample on the sample stage. There is a need to do. The optical imaging apparatus according to this embodiment is designed to reduce such time and effort.

  In the optical imaging apparatus of this embodiment, the sample stage 5 is divided into two of an X axis and a Y axis that are parallel to the mounting surface on the sample stage 5 and orthogonal to each other by a sample stage moving mechanism 30 including a motor, a slide rail, and the like. It can move in the axial direction. The sample stage drive unit 31 operates the motor included in the sample stage moving mechanism 30 in response to a control signal from the control unit 32, thereby moving the sample stage 5 to any position on the X axis and the Y axis (for example, in FIG. 5). To the position 5 ′). Since the position of the photometry unit A including the laser light source unit 1, the emission unit 3, the reflecting mirror 4, the objective lens 10, the spectroscopic unit 11, the fluorescent photographing camera 12 and the like is fixed, the sample stage 5 is moved as described above. Then, the irradiation range of the excitation light changes on the lower surface of the biological sample 7 placed on the sample stage 5 (or 5 ′), and the imaging range of the emitted excitation light intensity changes accordingly.

In the optical imaging apparatus of the present embodiment, a fluorescence intensity distribution image over a wide range of the biological sample 7 can be obtained as follows.
That is, the control unit 32 controls the sample stage driving unit 31 so that the sample stage 5 comes to a predetermined initial position, and the sample stage driving unit 31 includes a drive signal in the sample stage moving mechanism 30 accordingly. Send to motor. As a result, the sample stage 5 is set to the initial position. In this state, the fluorescence imaging camera 12 captures a fluorescence intensity distribution image based on the fluorescence that has arrived from the biological sample 7, and the obtained image data is used as an image processing unit. Send to 33. The image processing unit 33 temporarily stores the received image data in the image memory 331.

  When one imaging is completed, the control unit 32 controls the sample stage driving unit 31 so that the sample stage 5 moves by a predetermined distance along the X axis and / or the Y axis, and after the sample stage 5 has moved. The fluorescence imaging camera 12 acquires a fluorescence intensity distribution image again. By repeating such an operation a plurality of times over a predetermined range, a predetermined range wider than a single imaging range is imaged without omission. When such imaging is completed, in the image processing unit 33, the image composition processing unit 332 reads out image data for different imaging ranges from the image memory 331, and for example, determines all parts of the image that have been captured by overlapping some images. A fluorescence intensity distribution image corresponding to a predetermined range is created by joining. Then, this image is displayed on the monitor 34. Of course, the obtained images may be drawn on the monitor 34 without connecting the images.

  FIG. 6 shows the configuration of the main part of an optical imaging apparatus according to another embodiment that enables similar imaging. In the embodiment shown in FIG. 6, the position of the sample stage 5 is fixed, and the entire photometry unit A can be moved in the biaxial directions of the X axis and the Y axis by the photometry unit moving mechanism 40. Then, under the control of the control unit 42, the photometry unit driving unit 41 operates a motor included in the photometry unit moving mechanism 40, so that the photometry unit A can be moved to any position on the X axis and the Y axis (for example, FIG. 6). The position is moved to the position of the symbol A ′ in the middle. Even with this configuration, it is possible to change the position of the imaging range in the biological sample 7 placed on the sample stage 5, and it is obvious that imaging similar to that in the embodiment shown in FIG. 5 is possible. is there.

  5 and 6 both automatically perform photographing over a wider range than the photographing range by the fluorescent photographing camera 12, but not automatically, depending on the operation by the measurer. The shooting range may be set. For example, in the configuration of FIG. 5, when a measurer operates the operation unit attached to the control unit 32, a control signal is sent from the control unit 32 to the sample stage driving unit 31 according to the operation amount and the like. Accordingly, the sample stage drive unit 31 operates a motor included in the sample stage moving mechanism 30 to move the sample stage 5 by a predetermined amount along the X axis and / or the Y axis. When the sample stage 5 moves relative to the photometric unit A, the position of the imaging range changes, so that the position of the fluorescence intensity distribution image displayed on the monitor 34 changes. Can be observed.

Furthermore, as a simpler configuration, the sample stage moving mechanism 30 and the photometric unit moving mechanism 40 do not have a driving source such as a motor, and the measurer rotates and slides the knob (knob). Depending on the driving force, the sample stage 5 and the photometry unit A may be moved.
5 and 6, the sample stage 5 or the photometry unit A is moved two-dimensionally, but it may be configured to move only in one axis direction.

  In addition, any of the above-described embodiments is merely an example of the present invention, and other than the above-described modified examples, any appropriate modification, correction, or addition within the spirit of the present invention is included in the scope of the claims of the present application. It is clear.

DESCRIPTION OF SYMBOLS 1 ... Laser light source unit 2 ... Optical fiber 3, 3a, 3b, 3c, 3d ... Ejection unit 4 ... Reflection mirror 5 ... Sample stand 5a ... Transparent board 5b ... Support stand 6 ... Opening window 7 ... Biological sample 8 ... Visible light emission part DESCRIPTION OF SYMBOLS 10 ... Objective lens 11 ... Spectroscopic unit 12 ... Fluorescence imaging camera 13 ... Visible imaging camera 14 ... Optical filter 20 ... Outer housing 21 ... Sample mounting upper cover 30 ... Sample stage moving mechanism 31 ... Sample stage drive part 32, 42 ... Control unit 33 ... Image processing unit 331 ... Image memory 332 ... Image composition processing unit 34 ... Monitor 40 ... Metering unit moving mechanism 41 ... Metering unit driving unit

Claims (8)

In an optical imaging apparatus that irradiates a biological sample with excitation light and detects fluorescence obtained from the biological sample to form a two-dimensional image,
a) a housing having an upper lid that is swingable on the upper rear side around the horizontal axis at the front of the upper surface;
b) a sample stage for placing the biological sample substantially horizontally on the upper surface, exposed when the upper lid is swung and opened to the upper rear side without any obstacles above it ; A sample stage provided with a light transmitting portion through which light is transmitted on the mounting surface;
c) including a plurality of emission units that emit excitation light irradiated on the biological sample placed on the sample table, and each of the plurality of emission units spatially transmits the excitation light beam using an optical refracting element. An illumination unit that emits while diverging,
d) an imaging unit that acquires an image of fluorescence emitted from the biological sample with respect to excitation light by the illumination unit;
e) guiding the excitation light so that excitation light from the illumination unit is irradiated from below onto the biological sample through the light transmission part of the sample stage, and emitting fluorescence emitted downward from the biological sample through the light transmission part. A light guide optical system including a common reflecting optical unit disposed in a space below the sample stage, which bends the excitation light and the fluorescence together in space to guide the imaging unit;
An optical imaging apparatus comprising: The optical imaging apparatus according to claim 1,
The plurality of emission units are arranged to surround an optical axis of fluorescence from the reflection optical unit toward the imaging unit, and are substantially coaxial with the optical axis of fluorescence from the biological sample to the reflection optical unit. An optical imaging apparatus, wherein the biological sample is irradiated with excitation light from an optical unit. The optical imaging apparatus according to claim 1 or 2,
It further includes an optical dividing unit that divides fluorescence from the biological sample into light of a plurality of wavelength ranges, and the imaging unit acquires images of light of a plurality of wavelength ranges divided by the optical dividing unit, respectively. An optical imaging apparatus comprising a plurality of imaging units. The optical imaging apparatus according to any one of claims 1 to 3,
The optical imaging device, wherein the optical refractive element includes a lens array. The optical imaging apparatus according to any one of claims 1 to 4,
A movement that causes the photometric unit including the illumination unit, the light guide optical system, and the imaging unit, and the sample stage to move relative to each other in a one-dimensional or two-dimensional manner parallel to the mounting surface of the sample stage. A mechanism part;
An operation unit that is operated by a measurer so that the photometric unit and the sample stage are in a desired positional relationship by the moving mechanism unit;
In addition, the measurement person can acquire a two-dimensional image by light obtained from an arbitrary part of the lower surface of the biological sample placed on the sample stage by performing an operation using the operation unit. An optical imaging apparatus characterized by the above. The optical imaging apparatus according to claim 5,
The moving mechanism unit includes a drive source,
An optical imaging apparatus, further comprising: a control unit that controls driving of the drive source in accordance with an operation of the operation unit by a measurer. The optical imaging apparatus according to any one of claims 1 to 4,
A movement that causes the photometric unit including the illumination unit, the light guide optical system, and the imaging unit, and the sample stage to move relative to each other in a one-dimensional or two-dimensional manner parallel to the mounting surface of the sample stage. A mechanism part;
An imaging control unit for acquiring images of fluorescence from the biological sample by the imaging unit a plurality of times while moving at least one of the photometry unit or the sample stage in a one-dimensional or two-dimensional manner by the moving mechanism unit;
An optical imaging apparatus further comprising: The optical imaging apparatus according to claim 7,
An image forming unit that reproduces a two-dimensional image in a wider range than a range obtained by one shooting by the imaging unit by combining a plurality of images acquired under the control of the shooting control unit. An optical imaging apparatus.

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