JP5077824B2 - Confocal microscope system - Google Patents

Description

  The present invention relates to a confocal microscope system in which an imaging direction of an image incident on a digital camera or the like is changed by switching an optical path.

In a conventional confocal microscope system, a confocal scanner unit (hereinafter referred to as CSU) is attached to an observation connection port of a microscope, and excitation light (generally using laser light) is irradiated to the sample through a pinhole. The fluorescence emitted from the light is captured by a detector. Since only point information can be obtained as it is, a laser beam is scanned two-dimensionally to capture surface information (not shown) but a two-dimensional information is captured via an image sensor (camera). Is taken in.

  FIG. 6 is a configuration explanatory view showing an example of a conventional confocal microscope system.

The microscope 1 is a sample observation fluorescent microscope. The CSU 2 is a confocal scanner unit that is attached to an observation connection port of the microscope 1 and includes two optical paths, a bright-field optical path 21 and a confocal optical path 22 inside. The laser light source 3 is an excitation light source. The optical fiber 4 introduces excitation light from the laser light source 3 into the CSU 2. The imaging device 5 is composed of a CCD camera or the like, and detects one of the fluorescent images of the sample emitted from the two optical paths of the bright field optical path 21 and the confocal optical path 22 of the CSU 2. The image capturing device 6 includes a personal computer, software, a board, and the like, and captures image data from the image sensor 5. The image capturing device 6 displays a fluorescent image of the sample on the display based on the captured image data.

  The operation of the FIG. 6 system will now be described.

When confocal observation is performed (the optical path is on the confocal side), the excitation light emitted from the laser light source 3 is introduced into the CSU 2 through the optical fiber 4 and irradiated onto the sample through a pinhole (not shown). . The fluorescence emitted from the excited sample is detected by the image sensor 5 via the confocal optical path 22 (passing through the pinhole) of the CSU 2. The image data output from the image sensor 5 is captured by the image capturing device 6 and the image is displayed on the display.

When performing bright field observation (the optical path is on the bright field side), the image observed by the microscope 1 is detected by the imaging device 5 via the bright field optical path 21 of the CSU 2 without passing through the pinhole. The image data output from the image sensor 5 is captured by the image capturing device 6 and the image is displayed on the display. Since the bright field optical path 21 provided separately from the confocal optical path 22 does not pass through the pinhole, an observation image of an optical microscope, a fluorescence microscope, an interference microscope, or the like can be obtained as it is without reducing the amount of light.

  The following patent documents are known as prior art of the confocal microscope system.

JP 2002-062480 A

In the confocal microscope system described above, images of an optical microscope, a fluorescence microscope, an interference microscope, and the like can be observed in the bright field optical path 21, so that each is compared with the confocal image from the confocal optical path 22. In this case, in addition to the comparison during observation (Live), a comparison is made with the image once captured. Normally, during observation (LIVE), either a bright-field image or a confocal image is displayed, and after the image being observed is captured, both are compared and displayed side by side.

In the case of a confocal microscope, a bio-related target is a living cell, and if the laser is irradiated too much, the cell becomes weak. In order to avoid this as much as possible, the approximate position of the cell is first searched for in the bright field optical path, and then switched to the confocal optical path for observation.

  By the way, if the imaging direction of an output image differs in each optical path of said confocal microscope system, the user cannot perform an image comparison in real time.

For example, in the system of FIG. 6, when an erect image is output from the bright field optical path 21 and an inverted image (rotated 180 degrees) is output from the confocal optical path 22 with respect to the input image to the CSU 2, Since the focus image is reversed, it is necessary to rotate the image every time one image is captured. That is, in order to compare the live data of both optical paths, the image software of the image capturing device 6 must be operated 180 degrees every time the optical paths are switched, which is very complicated. .

  The present invention has been made to solve the above-described problems of the prior art, and an object thereof is to realize a confocal microscope system that does not require operation of image processing software and can compare images in real time without any particular awareness of the user. It is said.

In order to achieve such a problem, the invention according to claim 1 of the present invention is:
In a confocal microscope system that captures an image whose imaging direction is changed by switching a plurality of optical paths into an image capturing device via an imaging unit,
An image to be output to the image capturing device by performing at least one of inversion and rotation on the image data output from the imaging unit corresponding to the switching so that the images by the plurality of optical paths are in the same imaging direction. Equipped with a rotating unit ,
The plurality of optical paths include a confocal optical path and a bright field optical path .

The invention according to claim 2
The confocal microscope system according to claim 1.
The image rotation unit includes:
A mapping processing unit that sequentially generates pixel coordinates of image data from the imaging unit;
And a rotation processing unit that changes at least one of a reading start position and a reading direction of the mapped data.

The invention described in claim 3
The confocal microscope system according to claim 1 or 2 ,
The image rotation unit rotates at least one of 90 °, 180 °, 270 °, upside down, left / right inversion, left / right inversion + 90 ° rotation, left / right inversion + 270 ° rotation, and through with respect to the image data output from the imaging unit. The process is performed.

According to the confocal microscope system of the present invention, in the confocal microscope system that captures an image whose imaging direction is changed by switching of a plurality of optical paths into an image capturing device via an imaging unit, the plurality of the plurality of optical paths are corresponding to an optical path switching signal. Image rotation for processing the image data output from the imaging means in at least one of up / down and left / right directions and rotation and outputting to the image capturing device so that the images in the optical path are in the same imaging direction By providing the unit, it is possible to realize a confocal microscope system that does not require operation of image processing software and can compare images in real time without any particular awareness of the user.

  Hereinafter, embodiments of the present invention will be described in detail with reference to the drawings.

FIG. 1 is an example of a confocal microscope system according to an embodiment of the present invention, and is a configuration explanatory view showing a case where there are two optical paths as in FIG. The same elements as those in FIG.

  The image rotation unit 7 is connected between the image sensor 5 and the image capturing device 6, and corresponds to the switching of the plurality of optical paths 21 and 22 so that the images of the plurality of optical paths 21 and 22 are in the same imaging direction. 5 is a unit for digitally processing the image data output from 5. The processing content is switched by an optical path switching signal output from the CSU 2 by at least one of inverting and rotating the image vertically and horizontally.

  FIG. 2 is a processing block diagram relating to processing performed by a CPU (not shown) in the image rotation unit 7.

The image data memory 71 is a storage means in which image data from the image sensor 5 is written. The mapping processing unit 72 sequentially generates pixel coordinates (x, y) corresponding to the pixels of the image data memory 71 when the screen is scanned line by line from the upper left to the lower right, thereby obtaining an image from the image sensor 5. Mapping data. The rotation processing unit 73 changes the coordinates (x, x, x) generated by the mapping processing unit 72 by changing at least one of the reading start position (coordinates) and the reading direction of the mapped data in accordance with the set processing content. Rotate or flip y) to generate new coordinates (x, y). The remapping processing unit 74 corresponds to the new array of pixels generated by the rotation processing unit 73, and the coordinates (x, y) when the screen is scanned line by line from the upper left to the lower right as described above. Generate sequentially. The image data transfer unit 75 sequentially reads out the image data remapped by the remapping processing unit 74 and transfers it to the image capturing device 6.

  Next, the operation of the image rotation unit 7 in FIG. 2 will be described. In FIGS. 2A to 2I, images at each processing stage are shown using an image in which an R character is captured for easy understanding.

First, as an example of processing, a case where the optical path switching signal rotates 90 degrees to the right is shown. Image data (FIG. 2A) output from the image sensor 5 is stored in the image data memory 71. Image data is read from the image data memory 71 so as to scan the screen line by line from the upper left to the lower right (FIG. 2B), and the mapping processing unit 72 correspondingly coordinates (x, y). Are sequentially generated (FIG. 2C). In the case of a screen of 1000 horizontal dots and 1000 vertical dots, coordinates (0, 0), (1, 0)... (1000, 0), (0, 1). This image data is read out from the coordinate (0, 1000) to the lower right coordinate (1000, 0) by the rotation processing unit 73, and becomes a pixel array rotated right by 90 degrees (FIG. 2D). Corresponding to the image data rotated 90 degrees to the right, coordinates (x, y) when the screen is scanned again line by line from the upper left to the lower right by the remapping processing unit 74 are sequentially generated (FIG. 2 (e)). ). The remapped image data is sequentially read out by the image data transfer unit 75 and transferred to the image capturing device 6 (FIG. 2 (f)). As a result, an image rotated 90 degrees to the right by the image capturing device 6 is displayed (FIG. 2 (g)).

Usually, inside the CSU 2, the image is inverted 180 degrees by a relay lens used in a pair configuration, and the rotation changes depending on the number of mirrors. In the combination of the mirror and the lens inside the CSU 2 in FIG. 1, the number of mirrors in both optical paths is the same four, and the lens configuration is four bright-field optical paths (2 × 2), so 180 ° rotation × 2 = A 360 degree rotation results in an erect image, and since there are two confocal optical paths, an inverted image rotated 180 degrees. FIG. 3A shows a bright field image (left side) and a confocal image (right side) in a conventional system that does not have such an image rotation unit.

In this case, when image data based on the image acquired from the confocal optical path 22 is processed by the image rotation unit 7, the rotation processing unit 73 starts from coordinates (1000, 1000) as shown in FIG. By reading out toward the lower right coordinate (0, 0), image data in which an inverted image (rotated 180 degrees) from the confocal optical path 22 is an erect image can be sent to the image capturing device 6. Accordingly, when the bright-field optical path 21 is selected as the optical path switching signal, the pixel is moved from the coordinates without rotation (from the coordinates (0, 0) to the coordinates (1000, 1000) at the lower right in the rotation processing unit 73). In the case where the image data is sent to the image capturing device 6 and the optical path switching signal selects the confocal optical path 22, the image data obtained by converting the inverted image (rotated 180 degrees) to the erect image as described above is used. Since the images are sent to the image capturing device 6, as shown in FIG. 3B, the imaging directions of the bright field image (left side) and the confocal image (right side) can be matched.

According to the confocal microscope system as described above, the image data output from the image sensor 5 is rotated by 180 degrees by the image rotation unit 7 so that the display image is captured in the same direction even when the optical path is switched. Therefore, the operation of the image processing software that has been performed every time the optical path is switched is unnecessary, and a confocal microscope system that can compare images in real time without any particular awareness of the user can be realized.

  Further, in the image capturing device 6, it is possible to superimpose and observe the bright field image and the confocal image.

In the system shown in FIG. 1, if necessary, the bright field image may be rotated 180 degrees through the confocal image.

Further, as shown in FIG. 2 (i), the rotation processing unit 73 can read out from the coordinate (1000, 0) toward the lower right coordinate (0, 1000) to rotate 270 degrees.

Further, by changing the direction of reading the coordinates in the rotation processing unit 73, it is possible to invert vertically and horizontally.

In the image rotation unit 7, processing such as 90 ° rotation, 180 ° rotation, 270 ° rotation, upside down, left / right inversion, left / right inversion + 90 ° rotation, left / right inversion + 270 ° rotation, through, etc. can be set. Even if the number of mirrors or lenses changes, it can be handled. These settings and setting changes are performed by a rotary select switch, a communication command, or the like.

In addition, the rotation processing unit 73 can erect an image tilted with respect to the image sensor by reading the coordinates in an oblique coordinate direction.

The rotation data processing may be set for the input image independently for each optical path.

In addition, each setting of the image rotation unit 7 may be automatically switched by an optical path switching signal associated with processing contents of the image rotation unit 7 in advance.

FIG. 4 is a configuration explanatory diagram illustrating a case where another optical path is provided outside the CSU in the second example of the confocal microscope system according to the embodiment of the present invention. The same elements as those in FIG.

  The CSU 2 </ b> A is a confocal scanner unit that is attached to the observation connection port of the microscope 1 and includes a confocal optical path 22. The external optical path 8 is a bright-field optical path provided outside the CSU 2A, and has a structure capable of switching the optical path with the confocal optical path 22 by an optical path switching unit (not shown). The image sensor 5 detects a fluorescence image of the sample emitted from the two optical paths of the external optical path 8 and the confocal optical path 22. The processing content of the image rotation unit 7 is switched by an optical path switching signal output from the optical path switching unit.

The processing operation by the image rotation unit 7 in the system of FIG. 4 is the same as that of FIG.

According to the confocal microscope system as described above, even when an external optical path is attached outside the CSU option, it is not necessary to operate the image processing software every time the optical path is switched. A confocal microscope system capable of comparing images in real time can be realized.

1 to 4 show the case where there are two optical paths. However, the present invention is not limited to this, and any number such as a case where a bright field optical path in the CSU and an external bright field optical path are used in combination is used. This can be applied to the case where the optical path is switched.

FIG. 5 is another configuration example of the image rotation unit 7 and is a configuration explanatory diagram showing an image imaging system used for the purpose of rotating an image from a camera, not limited to a confocal scanner system. The same elements as those in FIG.

  Since the mounting direction of the constituting camera is limited, an image signal obtained by rotating the input image 90 degrees to the right is output from the image sensor 5, but image capture is performed by setting the image rotation unit 7 as 270 degrees rotation. In the apparatus 7, an upright image is obtained.

According to the image pickup system as described above, the mounting direction of the camera can be changed in various ways by changing the rotation setting of the image rotation unit. Therefore, it is effective when the mounting direction is limited by various camera shapes.

It is also effective for correcting the image direction when the image data is reversed on the image capture board.

It is composition explanatory drawing which shows one Example of the confocal microscope system which concerns on embodiment of this invention. FIG. 6 is a processing block diagram showing processing performed in the image rotation unit 7. It is operation | movement explanatory drawing which shows the bright-field image and confocal image in a conventional system and a FIG. 1 system. It is composition explanatory drawing which shows the 2nd Example of the confocal microscope system which concerns on embodiment of this invention. FIG. 10 is a configuration explanatory view showing another application example of the image rotation unit 7. It is composition explanatory drawing which shows an example of the conventional confocal microscope system.

Explanation of symbols

5 Imaging means 6 Image capturing device 7 Image rotation unit 8, 21 Bright field optical path 22 Confocal optical path 72 Mapping processing unit 73 Rotation processing unit

Claims (3)

In a confocal microscope system that captures an image whose imaging direction is changed by switching a plurality of optical paths into an image capturing device via an imaging unit,
An image to be output to the image capturing device by performing at least one of inversion and rotation on the image data output from the imaging unit corresponding to the switching so that the images by the plurality of optical paths are in the same imaging direction. Equipped with a rotating unit ,
The confocal microscope system, wherein the plurality of optical paths include a confocal optical path and a bright field optical path . The image rotation unit includes:
A mapping processing unit that sequentially generates pixel coordinates of image data from the imaging unit;
The confocal microscope system according to claim 1, further comprising: a rotation processing unit that changes at least one of a reading start position and a reading direction of the mapped data. The image rotation unit rotates at least one of 90 °, 180 °, 270 °, upside down, left / right inversion, left / right inversion + 90 ° rotation, left / right inversion + 270 ° rotation, and through with respect to the image data output from the imaging unit. confocal microscopy system according to claim 1 or 2, characterized in that the processing.