JP2009036764A - Two-wavelength simultaneous observation optical device - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide an inexpensive two-wavelength simultaneous observation optical device with a simple structure which is small, easy to handle, and easy even for a user with little knowledge of optics to conduct two-wavelength observation using a fluorescence microscope. <P>SOLUTION: The two-wavelength simultaneous observation optical device B includes a two-wavelength separation mirror box 10 for simultaneously observing fluorescent light of two different wavelengths emitted from an observation object, two CCD cameras 11, 13, and a monitor divider. Chromatic aberration generated by the two different wavelengths fluorescent light is projected on the respective two CCD cameras 11, 13. Accordingly, respective optical path distances can be finely adjusted and be corrected by spacers 12, 14, which are attached on the fluorescence microscope to be used. <P>COPYRIGHT: (C)2009,JPO&INPIT

Description

  The present invention relates to a microscope technique, and more particularly to a two-wavelength simultaneous observation optical apparatus that enables simultaneous observation of two types of fluorescence emitted from a sample and enables the miniaturization of the microscope.

  Conventionally, in a fluorescence microscope, a technique in which a filter is provided in an optical path between a light source and a camera to enable two-wavelength observation is known (see Patent Documents 1 to 4).

  The two-wavelength observation in the fluorescence microscope is performed by arranging a filter wheel in the optical path between the light source and the camera and synchronously controlling the filter wheel.

  On the other hand, there is a commercially available two-wavelength simultaneous observation optical unit that separates two fluorescent lights by a dichroic mirror and then separately projects them again onto one CCD light receiving surface.

JP-A-8-234110 Japanese Patent Application Laid-Open No. 11-072711 JP-T-2001-521422 JP 2006-038940 A

  The above-described conventional technique of synchronously controlling two filter wavelengths by synchronously controlling a filter wheel cannot be used for high-speed shooting at a video rate because the control system takes 500 milliseconds or more to replace the filter. Furthermore, software and a personal computer that control both the shutter control system and the moving image recording system are required.

  In the conventional two-wavelength simultaneous observation technique, the optical unit has a single light-receiving surface, so that it is inevitable that the apparatus is complicated and enlarged in order to eliminate chromatic aberration caused by two different fluorescences. Moreover, in the conventional two-wavelength simultaneous observation technique, the optical unit has a single light-receiving surface, so when the fluorescence intensities of two different fluorescences are greatly different, it is impossible to adjust the sensitivity of the CCD camera according to each fluorescence intensity. It is.

  An object of the present invention is to solve the above-described problems of the prior art, and an object thereof is to realize a two-wavelength simultaneous observation optical device that has a simple structure and can be miniaturized.

  In order to solve the above-described problems, the present invention is a two-wavelength simultaneous observation optical apparatus that is used by being attached to a fluorescence microscope, and is a two-wavelength separation for simultaneously observing two different wavelengths of fluorescence emitted from an observation target Provided is a two-wavelength simultaneous observation optical device characterized by a configuration including a mirror box, two CCD cameras, and a monitor divider.

  Chromatic aberrations caused by the two different wavelengths of fluorescence are projected separately on the two CCD cameras, and therefore, it is preferable to correct the light path distance by finely adjusting the optical path distance with a spacer.

  The two-wavelength separation mirror box may be a mirror box equipped with a detachable dichroic mirror, and the mirror box may be replaced so that the two-wavelength separation mirror box can cope with the combined wavelength.

  The image from the two CCD cameras may be integrated by using a monitor divider.

  It is good also as a structure which added the mirror box containing the mirror further to the said mirror box.

  A two-wavelength simultaneous observation optical device characterized in that it is simple and compact without using a relay lens system.

According to the two-wavelength simultaneous observation optical apparatus according to the present invention, the following remarkable effects are produced.
(1) Two different fluorescences can be easily observed simultaneously.
(2) Compared with a conventional two-wavelength simultaneous observation optical apparatus, a user who is small in size and easy to handle and has little optical knowledge can easily perform two-wavelength observation using a fluorescence microscope.

(3) The component parts of the two-wavelength simultaneous observation optical device of the present invention can be provided at low cost because commercially available products may be used.
(4) Since it is possible to record and observe images simply by connecting to a commercially available video or monitor, it is not necessary to purchase control software or image editing software separately.

  The best mode for carrying out the two-wavelength simultaneous observation optical apparatus according to the present invention will be described below with reference to the drawings based on the embodiments.

  The feature of the two-wavelength simultaneous observation optical device of the present invention is that two CCD cameras are connected to a two-wavelength separation mirror box including a dichroic mirror, and a fluorescent image having two different wavelengths emitted from an observation target is divided by the dichroic mirror. In addition, each fluorescent image is configured to be projected onto two CCD cameras separately.

  In this way, since the images having the fluorescence wavelengths are separated and projected onto the two CCD cameras, the images having different fluorescence intensities are projected onto the respective CCD cameras, so that each fluorescence image can be taken with optimum sensitivity. It becomes possible.

  Then, the images from the respective CCD cameras are integrated as one image by the monitor divider and image recording is performed. Further, by adding a mirror box including a mirror, both the mirror image and the normal image generated by the division of the dichroic mirror can be converted into a mirror image or a normal image.

  Examples 1 to 3 of the two-wavelength simultaneous observation optical device according to the present invention will be described below with reference to FIGS.

  FIG. 1 is a diagram for explaining a first embodiment of the two-wavelength simultaneous observation optical apparatus according to the present invention. The wavelength simultaneous observation optical device B of the present invention is applied to the fluorescence microscope A. The configuration of the fluorescence microscope A shown in FIG. 1 is a basic configuration of the fluorescence microscope to the last.

  In the fluorescence microscope A, light incident from the light source 1 becomes excitation light limited to a specific wavelength by the excitation filter 2. This excitation light is reflected by the dichroic mirror 5 to excite the observation object 3 via the objective lens 4, and the fluorescent image emitted therefrom is transmitted by the dichroic mirror 5 and the absorption filter 6 and then reflected by the mirror 7. The two-wavelength simultaneous observation optical apparatus B of the present invention is introduced.

  The variable spacer 8 is used to form an image of the fluorescence from the fluorescence microscope A on the light receiving surfaces of the CCD cameras 11 and 13 under optimum conditions, and the distance between the fluorescence microscope A and the two-wavelength simultaneous observation optical device B can be adjusted. It is a variable spacer.

  The mirror box 10 containing the dichroic mirror 9 can be easily replaced. Two-wavelength simultaneous observation of any combination is possible by replacing the mirror box equipped with a dichroic mirror matched to the target wavelength in advance.

  Each fluorescence divided by the dichroic mirror 9 mounted on the mirror box 10 forms an image on the light receiving surfaces of the CCD cameras 11 and 13, respectively. Chromatic aberration caused by two images having different wavelengths can be eliminated by changing the thicknesses of the variable spacers 12 and 14 disposed immediately before the CCD cameras 11 and 13, respectively.

  FIG. 2 is a diagram for explaining Example 1 of the two-wavelength simultaneous observation optical device according to the present invention. The two-wavelength simultaneous observation optical apparatus B of the second embodiment is applied to a fluorescence microscope A having the same configuration as that of the first embodiment. Similarly to the first embodiment, the variable spacer 7 forms an image of the fluorescence from the fluorescence microscope A on the light receiving surfaces of the CCD cameras 11 and 13 under optimum conditions.

  The characteristic configuration of the second embodiment is as follows. In FIG. 2, a mirror cube 16 including a mirror 15 is connected to the mirror box 10 so that a normal image transmitted through the dichroic mirror 9 installed in the mirror box 10 is a mirror image.

  Then, variable spacers 12, 14, and 17 are provided for adjusting the focus of the image projected on the CCD cameras 11 and 13 and eliminating chromatic aberration.

  FIG. 3 is a diagram for explaining a third embodiment of the two-wavelength simultaneous observation optical apparatus according to the present invention. The two-wavelength simultaneous observation optical apparatus B of Example 3 is applied to a fluorescence microscope A having the same configuration as that of Example 1. Similarly to the first embodiment, the variable spacer 8 forms an image of the fluorescence from the fluorescence microscope A on the light receiving surfaces of the CCD cameras 11 and 13 under optimum conditions.

  The characteristic configuration of the third embodiment is as follows. In FIG. 3, a mirror cube 19 including a mirror 18 for making a mirror image reflected from the dichroic mirror 9 installed in the mirror box 10 into a normal image is connected to the mirror box 10.

  Then, variable spacers 12, 14, and 20 are provided for adjusting the focus of the image projected on the CCD cameras 11 and 13 and eliminating chromatic aberration.

(Example of use)
FIG. 4 is a diagram for explaining an example of use of the two-wavelength simultaneous observation optical device 23 according to the present invention. Images from the two-wavelength simultaneous observation optical device connected to the side surface of the mirror body 21 such as a fluorescence microscope are integrated by a monitor divider 25 via a video cable 24 and recorded in a recording device (video, hard disk recorder) 26, and a monitor 27. It is configured to be projected on the screen. Reference numeral 22 denotes a light source.

  The best mode for carrying out the two-wavelength simultaneous observation optical apparatus according to the present invention has been described based on the embodiments. However, the present invention is not limited to such embodiments, and the scope of the claims is described. It goes without saying that there are various embodiments within the scope of the technical matters.

It is a figure explaining Example 1 of the two-wavelength simultaneous observation optical apparatus of this invention. It is a figure explaining Example 2 of the two-wavelength simultaneous observation optical apparatus of this invention. It is a figure explaining Example 3 of the two-wavelength simultaneous observation optical apparatus of this invention. It is a figure explaining the usage example of the two-wavelength simultaneous observation optical apparatus of this invention.

Explanation of symbols

DESCRIPTION OF SYMBOLS 1, 22 Light source 2 Excitation filter 3 Observation object 4 Objective lens 5 Dichroic mirror 6 Absorption filter 7, 15, 18 Mirror 8 Variable spacer with microscope 9 Dichroic mirror 10, 16, 19 Mirror box 11, 13 CCD camera 12, 14, 17, 20 Variable spacer 21 Mirror body such as a fluorescence microscope 22 Light source 23 Two-wavelength simultaneous observation optical device 24 Video cable 25 Monitor divider 26 Video / hard disk recorder 27 Monitor A Fluorescence microscope B Two-wavelength simultaneous observation optical device

Claims (6)

A two-wavelength simultaneous observation optical device used attached to a fluorescence microscope,
A two-wavelength simultaneous observation optical device comprising a two-wavelength separation mirror box for simultaneously observing two different wavelengths of fluorescence emitted from an observation object, two CCD cameras, and a monitor divider .   2. The chromatic aberration caused by the two different wavelengths of fluorescence is projected separately on the two CCD cameras, and is thus corrected by finely adjusting the optical path distance with a spacer. 2 wavelength simultaneous observation optical device.   3. The two-wavelength separation mirror box is a mirror box equipped with a detachable dichroic mirror, and the mirror box is exchanged so that the two-wavelength separation mirror box can be adapted to the wavelength of the combination. Two-wavelength simultaneous observation optical device.   2. The two-wavelength simultaneous observation optical apparatus according to claim 1, wherein the microscope images from the two CCD cameras are integrated by using a monitor divider.   The two-wavelength simultaneous observation optical device according to claim 1, wherein a mirror box including a mirror is further added to the mirror box.   A two-wavelength simultaneous observation optical device characterized in that it is simple and compact without using a relay lens system.

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