JP2010014964A - Living body observation apparatus - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a living body observation apparatus for capturing an image for detection and an image for observation using one image pickup device. <P>SOLUTION: This living body observation apparatus includes: an objective lens disposed close to a sample; an imaging device for imaging the sample; an objective lens driving device for driving the objective lens in the direction of correcting displacement of the sample; and a control device for controlling the imaging device to obtain an observation image of the sample and a detecting image for detecting the displacement of the sample. <P>COPYRIGHT: (C)2010,JPO&INPIT

Description

  The present invention relates to a living body observation apparatus.

In recent years, in biological research, so-called in vivo observation has been performed in which an individual experimental animal is used as a specimen and its organs and the like are observed with an optical microscope while alive. In in-vivo observation, the observation object has movements such as pulsation and respiratory movement, so that the observation position is shifted and the focus is likely to be blurred. In order to eliminate such displacement of the observation position and out-of-focus blur, an apparatus (for example, Patent Document 1) that tracks the entire microscope including the imaging unit in accordance with the movement of the observation target, or according to the movement of the observation target. For example, a device for tracking an objective lens (for example, Patent Document 2) is known. In Patent Document 2, a detection camera is provided separately from the observation camera in order to detect the movement of the observation target.
JP-A-7-222754 JP 2007-187810 A

  Usually, when observing a living body, the light source cannot be so strong. For this reason, the frame rate when imaging is reduced. FIG. 2 schematically shows the frame rate and the reading speed. The low speed reading in FIG. 2 means that the frame rate is small. In order to obtain an image for observation, it is generally necessary to perform low-speed reading. On the other hand, the high-speed reading in FIG. 2 means that the frame rate is high, and in order to detect the displacement of the living body, it is necessary to increase the frame rate in this way.

  Therefore, a conventional biological observation microscope apparatus includes two cameras for observation and detection having different frame rates. However, the use of two cameras has a problem in that the configuration becomes complicated and the cost increases.

  The present invention has been made in view of the above problems, and in a living body observation apparatus for observing a living body alive, it is possible to capture a detection image and an observation image with a single imaging apparatus, An object of the present invention is to provide a living body observation apparatus having a simplified configuration.

In order to achieve the above object, the present invention provides the following means.
The present invention includes an objective lens disposed close to a sample, an imaging device that images the sample via the objective lens, an objective lens driving device that drives the objective lens in a direction to correct the displacement of the sample, There is provided a living body observation apparatus including a sample observation image and a control device that controls the imaging device so as to acquire a detection image for detecting displacement of the sample.

  According to the present invention, it is possible to capture a detection image and an observation image with a single imaging device. Thereby, the structure of an observation apparatus can be simplified.

  In the above-described invention, the imaging device is an XY address type imaging device, and the control device can switch a readout pixel of the imaging device in imaging of an observation image and imaging of a detection image.

  In the above invention, the image pickup apparatus may include an image pickup device including long exposure pixels and short exposure pixels.

  In the above invention, the imaging device may include an imaging device in which the long-time exposure pixels and the short-time exposure pixels are arranged in every other column. Alternatively, in the imaging device, the long exposure pixels and the short exposure pixels may be alternately arranged.

In the above invention, the imaging apparatus may include an imaging device including two light-sensitive portions in one pixel and including pixels that can perform both high-speed reading and low-speed reading.
In the above invention, the imaging device may include an imaging element having a low-speed readout region and a high-speed readout region.
Moreover, in the said invention, the said control apparatus can control the said imaging device so that the imaging of the image for a detection by short-time exposure and the imaging of the image for an observation by long-time exposure may be performed alternately.

Further, the living body observation apparatus may be a living body observation microscope, or may be an endoscope or other observation apparatus.
The sample observed in the living body observation apparatus may be a cell, tissue, or internal organ in the living body, and the minute behavior in the living sample is observed by correcting the dynamic behavior of the living body. It is possible.

  ADVANTAGE OF THE INVENTION According to this invention, the imaging for a detection and the imaging for an observation can be performed with one imaging device, and the biological observation apparatus with the more simplified structure can be provided.

  The biological observation apparatus of the present invention is suitably used as a microscope observation apparatus, but is not limited to this, and can be used as, for example, an endoscope observation apparatus or other appropriate apparatus. Hereinafter, for convenience of explanation, a microscope observation apparatus will be described in detail as an example.

A microscope observation apparatus 1 according to a first embodiment of the present invention will be described with reference to FIG.
A microscope observation apparatus 1 according to this embodiment includes a stage 2 on which a sample S is mounted, an excitation light source 3 that emits excitation light E, and an illumination lens (illumination optical system) 4 that guides the excitation light E from the excitation light source 3. The excitation light E guided by the illumination lens 4 is guided to the sample S, while the objective lens 5 that collimates the fluorescence F generated in the sample S, and the fluorescence F collimated by the objective lens 5 from the excitation light E. A branching dichroic mirror 6 and an imaging device (light detection unit) 9 such as a CCD for detecting fluorescence F are provided. Reference numeral 20 in the figure denotes an imaging lens.

  The excitation light source 3, the illumination lens 4, the dichroic mirror 6, and the imaging device 9 are provided in a microscope body 10. On the other hand, the objective lens 5 is separated from the microscope body 10 and supported by the objective lens driving device 11.

  The objective lens driving device 11 includes an arm 12 that arranges the objective lens 5 between the stage 2 and the microscope main body 10, and a first drive mechanism 13 that moves the objective lens 5 held by the arm 12 in the vertical direction. And second and third drive mechanisms 14 and 15 for moving the objective lens 5 in two horizontal directions, and swinging the objective lens 5 about an axis along two directions orthogonal to the optical axis of the objective lens 5. The fourth and fifth drive mechanisms 16 and 17 are provided. The first to third drive mechanisms 13 to 15 include, for example, motors 13a to 15a and a linear movement mechanism such as a ball screw (not shown) connected to the motors 13a to 15a. The fourth and fifth drive mechanisms 16 and 17 include, for example, motors 16a and 17a and speed reduction mechanisms 16b and 17b connected to the motors 16a and 17a.

  The imaging device 9 processes the image acquired by the imaging device 9 to calculate the displacement amount and the direction of the sample S, and the sample S with respect to the driving mechanisms 13 to 17 of the objective lens driving device 11. A drive control device 18 for outputting a drive command for driving the objective lens 5 in a direction for correcting the deviation of the optical axis of the objective lens 5 due to the displacement is connected.

  The imaging device 9 is further connected to a control device 19 that controls the imaging device 9 to acquire a detection image and an observation image.

Next, the image sensor 23 provided in the imaging device 9 will be described.
The imaging device 9 is provided with an XY address type imaging device 23 such as a CMOS. An XY address image sensor can read a signal at an arbitrary position at an arbitrary timing. Here, the image pickup device 23 includes two types of pixels, a long-time exposure pixel 21 and a short-time exposure pixel 22. The long-time exposure pixel 21 refers to a pixel capable of low-speed readout as shown in FIG. On the other hand, the short-time exposure pixel 22 refers to a pixel capable of high-speed reading as shown in FIG.

  In the present embodiment, the control device 19 designates the pixels used when capturing the detection image and the observation image.

  In the first embodiment, the long exposure pixels 21 and the short exposure pixels 22 are arranged in every other row as shown in FIG. In the second embodiment, the long exposure pixels 21 and the short exposure pixels 22 are alternately arranged as shown in FIG.

  Furthermore, in the third embodiment, an image pickup device composed of pixels capable of both long time exposure and short time exposure is used. Such a pixel has two photosensitive parts for long exposure and short exposure. The wavelength for imaging differs between pixels that can be exposed for a long time and pixels that can be exposed for a short time.For example, a light sensitive part for long exposure and a light sensitive part for short time exposure are overlapped, Pixels that can be detected can be used.

  In each of the above embodiments, the long-time exposure pixel 21 and the short-time exposure pixel 22 may be different pixels suitable for the respective functions. Further, the same pixel may be used for the long-time exposure pixel 21 and the short-time exposure pixel 22 as long as the pixel has functional characteristics that satisfy both functions of long-time exposure and short-time exposure. The long exposure pixel 21 and the short exposure pixel 22 further include an optical filter suitable for the wavelength detected by each.

  In the microscope observation apparatus 1 according to the present embodiment configured as described above, as shown in FIG. 5, the objective lens 5 is moved in the Z-axis direction along the optical axis by the operation of the objective lens driving device 11, and the optical axis. Are movable in the X axis and Y axis directions orthogonal to each other, and in the A and B directions around the X axis and Y axis, respectively, and the objective lens 5 can follow the displacement of the living body.

The operation of the microscope observation apparatus 1 according to the present embodiment configured as described above will be described below.
In the microscope observation apparatus 1 according to the present embodiment, an image for detecting the displacement of the sample is captured using the short-time exposure pixels 22 in accordance with a command from the control device 19. Image information for detection acquired by the imaging device 9 using the short-time exposure pixels 22 is sent to the drive control device 18 and is subjected to image processing in the drive control device 18, so that the displacement direction, the displacement amount, and A swing angle or the like is detected. The drive control device 18 calculates command signals to be sent to the drive mechanisms 13 to 17 of the objective lens drive device 11 based on the detection result, and the drive mechanisms 13 to 17 are driven based on the command signals, so that the biological displacement is detected. Follow-up control is performed. On the other hand, in accordance with a command from the control device 19, an image for observation of the sample is acquired by the imaging device 9 using the long-time exposure pixel 21.

  In the present embodiment, the reading of the long-time exposure pixels 21 and the short-time exposure pixels 22 is controlled by the control device 19 and is performed independently and in parallel. Alternatively, the control device 19 may switch the readout pixels of the image sensor and pick up images alternately.

  When the long-time exposure pixels 21 and the short-time exposure pixels 22 are arranged in every other row as in the first embodiment, detection is performed because the higher the pixel density, the higher the detection resolution. It is preferable that the vibration direction be arranged in parallel with a column of the same kind of pixels.

  With the above-described configuration, both high-speed reading and low-speed reading can be performed with a single imaging device. Therefore, in the case of high-speed reading, for example, imaging can be performed at a frame rate of 1000 fps, and biological displacement can be detected with high accuracy. Further, in the case of low-speed reading, it is possible to take an image with long exposure, and a bright image with a good S / N ratio can be obtained.

  For detection of biological vibration, a marker that generates high-luminance light such as fluorescence or chemiluminescence is used. In this case, an optical filter having wavelength selectivity suitable for living body observation is attached to the long-time exposure pixel 21. Further, an optical filter having a wavelength selectivity suitable for marker detection is attached to the short-time exposure pixel 22. The wavelength to be acquired is selected by using an optical filter having wavelength selectivity suitable for each.

  Alternatively, natural texture pattern matching may be used without using a marker for detection of biological vibration. In this method, the observed image is patterned, and the movement amount (displacement amount) of the image between frames is obtained by matching (adjusting the deviation). According to this method, since it is not necessary to detect a marker, the same optical filter can be used for the long-time exposure pixel and the short-time exposure pixel. Therefore, the trouble at the time of manufacturing an image sensor can be simplified.

  Next, a microscope observation apparatus 1 according to a second embodiment of the present invention will be described with reference to FIG. The present embodiment has the same configuration as that of the microscope observation apparatus 1 according to the first embodiment described above, except that the configuration of the imaging element 24 included in the imaging device 9 is different.

  In the second embodiment, the imaging device 9 includes an imaging element 24 having a low-speed readout region 25 and a high-speed readout region 26. Preferably, a part of the image sensor 24 is a high-speed reading area 26 and the other area is a low-speed reading area 25. The low-speed reading area 25 may use long-time exposure pixels, and the high-speed reading area 26 may use short-time exposure pixels. However, the same pixel may be used in both regions as long as both the long-exposure imaging and the short-exposure imaging can be performed.

  In the present embodiment, a marker is detected in the high-speed reading area 26 and an observation image is acquired in the low-speed reading area 25. Therefore, the marker image can be excluded from the viewing field.

  With the configuration described above, it is possible to obtain a detection image and an observation image at the same time with a single imaging device, and obtain an observation image that is not affected by the marker.

  Next, a microscope observation apparatus 1 according to a third embodiment of the present invention will be described with reference to FIG. In the present embodiment, the imaging apparatus 9 has the same configuration as the microscope observation apparatus 1 according to the first embodiment, except that the imaging element included in the imaging apparatus 9 is different.

  In the third embodiment, an image sensor composed of the same pixels is used. This pixel is a pixel applicable to both high-speed reading and low-speed reading.

  In the microscope observation apparatus 1 according to the present embodiment, as shown in FIG. 7, an image for detecting a biological displacement is captured by short-time exposure according to a command from the control device 19. The obtained detection image is sent to the drive control device 18 as in the above embodiment, and the drive control device 18 performs image processing to detect displacement. The drive control device 18 calculates a command signal to be sent to the objective lens drive device 11 based on the detection result, and the drive mechanisms 13 to 17 are driven based on the command signal.

  Next, in accordance with a command from the control device 19, an observation image is taken by long exposure. While the obtained image is displayed on an observation system display unit (not shown), a further image for detecting a living body displacement is further captured. In this way, imaging by short exposure for detection and imaging by long exposure for observation are performed alternately. While an image for observation is being taken, follow-up control may be performed according to a displacement prediction based on a previously detected result of biological displacement.

  By doing in this way, even if it is an imaging device provided with the image sensor comprised from the single pixel similar to the past, the image for a detection and observation can be acquired with one imaging device. . Moreover, since all the pixels can be used for imaging, a highly accurate image can be acquired.

  Note that the imaging conditions may be changed between the short exposure and the long exposure. For example, the amount of light for detecting the marker may be increased during short-time exposure. These controls can be performed by the control device 19.

  The configuration and operation of the present invention have been described above using an upright microscope as an example, but the present invention is not limited to this and may be an inverted type. Further, the living body observation and the optical system for the marker may be a combination other than imaging by illumination light or fluorescence. The scope of the present invention is not limited to a microscope, and any observation apparatus including an imaging unit, an objective unit including an objective lens, and the objective unit driving apparatus is included in the present invention. Furthermore, the present invention can be applied not only to in vitro observation but also to any dynamic observation that enables tracking by combining two or more exposure times as described above.

The typical whole block diagram which shows the microscope observation apparatus which concerns on embodiment of this invention. The schematic diagram which shows the frame rate at the time of imaging. 1 shows an embodiment of an image sensor used in the observation apparatus of the present invention. The other Example of the image pick-up element used for the observation apparatus of this invention. The perspective view which shows the relationship between the objective lens and coordinate axis in description of the microscope observation apparatus of FIG. An example of the image pick-up element used for the observation apparatus which concerns on 2nd Embodiment. The schematic diagram which shows the imaging by the observation apparatus which concerns on 3rd Embodiment in time series.

Explanation of symbols

  S ... sample, E ... excitation light (light), F ... fluorescence (return light), 1 ... microscope observation device, 3 ... excitation light source (light source), 4 ... illumination lens (illumination optical system), 5 ... objective lens, 9 DESCRIPTION OF SYMBOLS ... Imaging device, 10 ... Microscope main body, 11 ... Objective lens drive device, 18 ... Drive control device, 19 ... Control device.

Claims (10)

An objective lens placed close to the sample;
An imaging device for imaging a sample through the objective lens;
An objective lens driving device for driving the objective lens in a direction for correcting the displacement of the sample;
A biological observation apparatus comprising: an observation image of the sample; and a control device that controls the imaging device so as to acquire a detection image for detecting displacement of the sample.   2. The image pickup device according to claim 1, wherein the image pickup device includes an XY address type image pickup device, and the control device switches a readout pixel of the image pickup device in image pickup for observation and image pickup for detection. The biological observation apparatus described.   The living body observation apparatus according to claim 1, wherein the imaging apparatus includes an imaging element including a long exposure pixel and a short exposure pixel.   The living body observation apparatus according to claim 3, wherein the imaging apparatus includes an imaging device in which the long-time exposure pixels and the short-time exposure pixels are arranged in every other row.   The living body observation apparatus according to claim 3, wherein the imaging apparatus includes an imaging element in which the long-time exposure pixels and the short-time exposure pixels are alternately arranged.   The image pickup apparatus includes an image pickup device including two light-sensitive portions in one pixel and composed of pixels capable of both high-speed reading and low-speed reading. 2. The biological observation apparatus according to 2.   The living body observation apparatus according to claim 1, wherein the imaging apparatus includes an imaging element having a low-speed reading area and a high-speed reading area.   2. The living body according to claim 1, wherein the control device controls the imaging device to alternately take a detection image by short-time exposure and an observation image by long-time exposure. Observation device.   The biological observation apparatus according to any one of claims 1 to 8, wherein the biological observation apparatus is a microscope for biological observation.   The living body observation apparatus according to claim 1, wherein the sample is a cell, tissue, or viscera in a living body.

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