KR101106633B1 - Dual vision optical molecular imaging device for real time fluorescence imaging of small animal - Google Patents

Abstract

The present invention relates to a fluorescence imaging technique that is widely used for molecular imaging research, and more specifically, it is possible to move and track a substance in a small animal by detecting a signal causing fluorescence using a fluorescent protein material or a dyeing material. The present invention relates to an optical molecular imaging device.

Fluorescent protein material, fluorescent phenomenon, small animal, LED light source

Description

Dual vision optical molecular imaging device for real time fluorescence imaging of small animal}

 The present invention relates to a fluorescence imaging technique that is widely used for molecular imaging research, and more specifically, the movement and tracking of a substance in a small animal by detecting and imaging a signal causing fluorescence using a fluorescent protein material or a dyeing material. This relates to a possible optical molecular imaging device.

 The fluorescent material is excited by external light and returns to the ground state to emit shifted light. According to the wavelength of the emitted light, there are various wavelengths ranging from representative wavelength bands such as green fluorescence, red fluorescence, and blue fluorescence to the near infrared region.

In relation to this, many fluorescent proteins are being developed worldwide and are mainly used for molecular imaging studies of small animals. Fluorescent protein refers to a protein that emits light when properly stimulated. Typically, fluorescent proteins, when irradiated with an excitation wavelength, emit light in the visible band, ie, light in the band of about 400 nm to 800 nm. By instrument operation, a wider band is also available.

Recently, as the research using molecular imaging technology has spread widely, various fluorescent proteins have been developed and reported for various purpose experiments. Molecular imaging through fluorescence signal detection is a useful tool that can be used continuously for drug development, cancer research, immunity and infection research.

As the latest technology, various materials for generating fluorescence, such as utilizing fluorescent materials obtained from living organisms, have been developed, and fluorescent materials in various wavelengths are coming out. Therefore, the expansion of imaging technology using various fluorescence phenomena is being made.

On the other hand, the fluorescence image has a number of disadvantages of a background signal, but the original signal has an advantage of being stronger by controlling the irradiation light source than the emission image. Therefore, in order to minimize the background signal of the fluorescent image and obtain a desired signal, a single wavelength band pass filter is used for exciting light and detection of a single wavelength.

In order to observe the fluorescence phenomenon, the researcher is interested in analyzing the location of fluorescence while observing the entire animal at once with in-vivo image, and the movement and reaction of the fluorescent protein in the blood vessel under the skin. To observe the phenomenon. Most of the area of the detection of fluorescence image signal can be largely divided into observing phenomena in small parts such as blood vessels by viewing or enlarging the whole, and there is a multi-wavelength detection technology that obtains various wavelength bands by separating fluorescence wavelength bands. .

Recently, techniques have been devised to use light emission to observe the internal behavior of animal host cells or tumor cells in a non-invasive way. For example, the PCT publication WO # 97/18841 describes lux operon to mitigate Salmonella bacteria, and this infectious agent has become a bioluminescent material. The process of infection can be tracked in intact animals without external stimulation of the luminescent material, but the observation required immobilization of the subject to provide sufficient image intensity. Therefore, the animal had to be imaged with a charge-coupled device on a two-stage microchannel enhancer head attached to a fully shielded box and attached to an Argus 50 image sensor. In this method, the subject is held stationary, the photon device measures the photon emission at the light-generating portion located on the subject until the photon emission image is established, and the image is passed through the opaque tissue of the animal mentioned. Had to detect.

While this known technique is useful, it is evident that a complex device is required to acquire fluorescence images and that many test subjects have not been evaluated quickly within a short period of time. Therefore, a more precise and easy-to-use device for obtaining fluorescence image and a device having various functions while quickly evaluating a large number of subjects in a short period of time are required.

The present inventors have completed the present invention as a result of trying to solve all the disadvantages and problems of the prior art as described above.

Accordingly, it is an object of the present invention to provide an optical molecular imaging apparatus capable of acquiring a desired portion more accurately in real time while simultaneously acquiring an image of a fluorescent material emitted from a small animal as a whole.

Another object of the present invention is to provide an optical molecular imaging device that can be used by selectively adjusting the LED light source and the white light source according to the function and characteristics.

Another object of the present invention is to provide an optical molecular imaging device that maximizes the user's convenience by using LabVIEW to observe the fluorescence in the small animal as a whole while at the same time the magnified observation is made in a selective position.

Another object of the present invention is to provide an optical molecular imaging device that can be conveniently used by reducing the user's operation by allowing one object to be observed in two images at the same time.

The objects of the present invention are not limited to the above-mentioned objects, and other objects that are not mentioned will be clearly understood by those skilled in the art from the following description.

In order to achieve the above object, the present invention is a housing forming a closed space; A movable stage installed under the housing and including a table on which a subject is located and a stage moving means for moving the table; Irradiation means for irradiating light for photographing the subject; And a plurality of photographing means for photographing the fluorescent material emitted from the subject as an image.

In a preferred embodiment, the plurality of photographing means includes total monitoring photographing means for photographing the fluorescence of the entire subject and magnification precision photographing means for photographing the fluorescence of the selected specific magnified portion.

In a preferred embodiment, the stage moving means is installed adjacent to the lower portion of the table includes a fine moving means for controlling to finely adjust the table automatically up and down, front and rear, left and right.

In a preferred embodiment, the stage moving means further comprises a stage height adjusting means installed below the micro moving means.

In a preferred embodiment, the movable stage further comprises a temperature control means for adjusting the temperature of the table.

In a preferred embodiment, the magnification precision photographing means includes a lens module for controlling the magnification of the optical image of the subject; A photographing module for photographing an optical image incident on the lens module; A detection filter wheel for detecting a fluorescence signal emitted from the subject; And a barrel in which the lens module is mounted at one end and a photographing module is mounted at the other end.

In a preferred embodiment, the irradiating means includes only the LED light source unit or together with the white light source unit.

In a preferred embodiment, when a plurality of specific positions are selected from the images captured by the whole monitoring photographing means, the specific positions of the subjects are photographed by the enlarged precision photographing means in the selected order after storing the selected positions. The stage moving means controls the table on which the subject is located so as to be at the position where it can be.

The present invention has the following excellent effects.

The optical molecular imaging device of the present invention can acquire an image of a fluorescent substance emitted from a small animal as a whole while simultaneously obtaining a desired portion more accurately in real time.

The optical molecular imaging device of the present invention can be used by selectively adjusting the LED light source and the white light source according to the function and characteristics.

The optical molecular imaging device of the present invention maximizes the user's convenience by using LabVIEW to observe the fluorescence in the small animal as a whole while at the same time expanding observation at a selective position.

The optical molecular imaging device of the present invention can be conveniently used by reducing the user's operation by allowing one object to be observed simultaneously with two images.

The terms used in the present invention were selected as general terms as widely used as possible, but in some cases, the terms arbitrarily selected by the applicant are included. In this case, the meanings described or used in the detailed description of the present invention are considered, rather than simply the names of the terms. The meaning should be grasped.

Hereinafter, with reference to the preferred embodiment shown in the accompanying drawings will be described in detail the technical configuration of the present invention.

However, the present invention is not limited to the embodiments described herein but may be embodied in other forms. Like reference numerals designate like elements throughout the specification.

1 is a perspective view of a small animal fluorescence image simultaneous observation optical molecular imaging device according to a preferred embodiment of the present invention, Figure 2 is an embodiment of a magnified precision photographing means and a white light source of the optical molecular imaging device shown in FIG. 3 is a detailed view according to an embodiment of a movable stage among the optical molecular imaging devices shown in FIG. 1.

Referring to FIG. 1, the small animal fluorescence image simultaneous observation optical molecular imaging apparatus 100 according to a preferred embodiment of the present invention includes a housing 110, a movable stage 120, an irradiation unit 130, and a plurality of photographing means ( 150, 160).

First, the housing 110 forms an enclosed space in which the main components of the optical molecular imaging apparatus 100 are installed. In particular, the housing 110 is configured to provide a space suitable for photographing, and is provided on one surface for convenience of photographing. It is preferable that an opening and closing door is formed.

Next, the movable stage 120 is installed under the housing 110 and serves to move the subject to a position most suitable for photographing, so that the table 121 and the table 121 on which the subject is located are moved. Preferably, the stage moving means 122 is included.

Looking at the configuration of the movable stage 120 in more detail with reference to Figure 3, the stage moving means 122 is installed adjacent to the lower portion of the table 121 automatically finely vertically up and down, front and rear, left and right It can be seen that it comprises a fine moving means (121a) for controlling to be adjustable. In particular, the fine movement means 121a is made of a left and right movement motor, a Shanghai movement motor and a front and rear movement motor, and the motors are automatically adjusted by a preset program to control the optimum focal length.

In some cases, the stage moving unit 122 may further include a stage height adjusting unit 122b. The stage height adjusting means 122b may be installed at the lower portion of the fine moving means 121a to be configured to move the height of the entire movable stage 120 greatly and to adjust the stage height adjusting means 122b. It may be configured manually but is not limited to being implemented automatically.

In addition, although not shown, the movable stage 120 may further include a temperature adjusting means having a known configuration capable of adjusting the temperature of the table 121 to maintain the body temperature of the experimental animal, which is a subject. In this case, it is preferable that the current temperature is displayed on the outside of the device and implemented so that the user can adjust the temperature.

Next, the irradiating means 130 serves to irradiate light for photographing a subject, and the irradiating means 130 may include only the LED light source 131 or may include the white light source 132 together. . In addition, in the case of including the LED light source unit 131 and the white light source unit 132, each light source may be used individually or combined.

The LED light source 131 may generally be composed of a blue LED and is preferably provided with an excitation filter having a special cutoff frequency that allows for an appropriate excitation wavelength, for example, an excitation frequency of 400 to 650 nm. In the present invention, a plurality of excitation filters may be provided in the form of a wheel. In particular, when an excitation filter of 435 nm, 468 nm, 488 nm, 544 nm, and 546 nm, each of which has a peak value of 5, has one more space, that is, 6 Wheels with two mounts. As such, when a space is present in the wheel, a light source that does not pass through the excitation filter can be irradiated to the subject. The beam width can also be determined by the size of the part to be observed.

The white light source unit 132 is configured to adjust brightness, and the white light source unit 132 also includes a wheel having a plurality of excitation filters and spaces that pass only a specific wavelength in the same way as the LED light source unit 131. As shown in FIG. 2, the present invention can be configured to connect the white light source unit 132 to enter the device from the outside by using fiber as an example. In particular, the white light source unit 132 and the LED light source unit 131 are light. The optical filter wheel 133 may be implemented to be connected by the connection portion 132a installed.

Next, the plurality of photographing means 140 serves to photograph the fluorescent material emitted from the subject as one or more whole monitoring photographing means 141 and one or more magnification precision photographing means 142.

The whole monitoring photographing means 141 is for photographing the entire experimental animal as a subject, and a photographing means having a known configuration including an image sensor such as a CCD or a CMOS may be used. And a fluorescence detection filter wheel 143 equipped with a fluorescence detection filter or a plurality of fluorescence detection filters. The entire monitoring photographing means 141 collects the image of the fluorescent material emitted from the experimental small animal, the collected image can be displayed on the monitor and stored digitally. In addition, image processing can be performed through standard software embedded in an external device, and can be implemented to continuously capture a high resolution image or acquire a video.

As shown in FIG. 2, the enlarged precision photographing means 142 includes an upright microscope including a lens module 142a and a barrel 142c, a photographing module 142b mounted on the top of the microscope, and a fluorescence detection filter wheel 143. It includes.

 The upright microscope including the lens module 142a and the barrel 142c performs the same function as a known microscope to allow a magnified observation of a specific part of the subject. In particular, since the lens module 142a serves as an objective lens of the microscope, the optical image of the subject plays a role in which the magnification is adjusted and is detachably implemented. For example, if a lens having a 2x to 10x magnification is attached and used according to a situation, the subject may be observed at different magnifications of the focal point.

The fluorescence detection filter wheel 143 is for observing the emission of the fluorescent material in the body of the experimental animal. In particular, in the present invention, as shown in FIG. 2, 540 nm, 544 nm, 549 nm, 610 nm, and 615 nm are respectively mounted and one blank is provided. Further, it is provided in the form of a wheel having six mounting parts, it is implemented to detect a light source that does not pass through the fluorescence detection filter.

The photographing module 142b is mounted on the other end of the barrel 142c, that is, the upper end of the microscope, and photographs an optical image incident on the lens module 142a. The photographing module 142b includes an image sensor such as a CCD or a CMOS. Images captured and collected by the photographing module 142b may be displayed on the monitor and stored digitally in the same manner as the entire monitoring photographing means 141. In addition, image processing can be performed through standard software embedded in an external device, and can be implemented to continuously capture a high resolution image or acquire a video.

Through such a configuration, the image captured by the entire monitoring photographing means 141 and the image photographed by the magnification precision photographing means 142 may be simultaneously displayed on a monitor to simultaneously observe in real time.

In addition, when the user selects a plurality of specific positions while viewing the images of all the experimental animals displayed on the monitor captured by the entire monitoring photographing means 141, the specific positions of the subjects are stored in the selected order and then the specific positions of the subjects are enlarged. It may be controlled to photograph by means 142. In this case, the driving of the stage moving means 122 may be controlled to move the table 121 on which the subject animal is placed to a position suitable for capturing the enlarged precision photographing means 142.

Simultaneous observation of small animal fluorescence images according to a preferred embodiment of the present invention configured as described above simplifies the process of observing a fluorescence image using a small animal into which a fluorescent substance such as a fluorescence protein is injected into a body In detail, first, a small experimental animal to be observed is placed in a live state under anesthesia, placed on a table 121 in the optical molecular imaging apparatus 100, and the height of the movable stage 120 is appropriately adjusted using the stage height adjusting means 122b. Adjust

After that, the light emitted from the LED light source unit 131 and / or the white light source unit 132 passes through the optical filter and is continuously exposed to the experimental animal lying on the table 121 to enter the photographing means for capturing an image. In this case, the fluorescent detection filter wheel 143 is used to capture the fluorescent material emitted from the experimental animal as an image having a desired wavelength, and the stage moving unit 122 is used to capture the desired location to select a desired position of the subject. In particular, the portion to be viewed can be enlarged and photographed by the lens module 142a of the magnification precision photographing means 142. The captured images are digitized and transferred to another external device for use, such as displayed on a monitor or processed for data storage or image processing via standard software built into the external device, continuous capture as high resolution images, or acquisition of video. Can be implemented.

As described above, according to the present invention, the image of the fluorescent substance emitted from the small animal can be obtained not only in the entire experimental animal but also at the same time in a more accurate manner, and the fluorescence can be observed in the small animal using LabVIEW. At the same time, magnified observation can be made at a selective position, and one object can be observed simultaneously in two images, thereby reducing the user's operation and maximizing user convenience.

As described above, the preferred embodiments have been illustrated and described, but are not limited to the above-described embodiments, and various modifications may be made by those skilled in the art without departing from the spirit of the present invention. And modifications will be possible.

1 is a perspective view of a small animal fluorescence image simultaneous observation optical molecular imaging device according to an embodiment of the present invention,

FIG. 2 is a detailed view according to an embodiment of a magnification precision photographing unit and a white light source unit of the optical molecular imaging apparatus shown in FIG. 1;

3 is a detailed view according to an embodiment of a movable stage of the optical molecular imaging device shown in FIG.

<Description of the symbols for the main parts of the drawings>

100: Simultaneous observation of small animal fluorescence images

110: housing 120: movable stage

121: table 122: stage moving means

122a: fine moving means 122b: stage height adjusting means

130: irradiation means 131: LED light source

132: white light source 133: optical filter wheel

140: recording means 141: total monitoring recording means

142: precision photographing means for magnification 142a: lens module

142b: camera module 142c: barrel

143: Detection filter wheel

Claims (8)

A housing defining an enclosed space; A movable stage installed under the housing and including a table on which a subject is located and a stage moving means for moving the table; Irradiation means for irradiating light for photographing the subject; And It includes a plurality of photographing means for photographing the fluorescent material emitted from the subject as an image, The plurality of photographing means includes total monitoring photographing means for photographing the fluorescence of the entire subject and magnification precision photographing means for photographing the fluorescence of the selected specific magnified portion. The magnification precision photographing means includes a lens module for controlling the magnification of the optical image of the subject; A photographing module for photographing an optical image incident on the lens module; A detection filter wheel for detecting a fluorescence signal emitted from the subject; And a barrel in which the lens module is mounted at one end and a photographing module is mounted at the other end. The lens module is detachably configured to use a lens having a desired magnification according to a photographing condition, and the detection filter wheel has a plurality of filters and spaces for passing only a specific wavelength for observation of emission of fluorescent material in the subject. Small animal fluorescence imaging simultaneous observation optical molecular imaging device, characterized in that. delete The method of claim 1, Small stage fluorescence image simultaneous observation optical molecular image, characterized in that the stage movement means is installed adjacent to the lower portion of the table to control the table to automatically adjust finely up, down, front, left and right Device. The method of claim 3, wherein And said stage moving means further comprises a stage height adjusting means installed under said micro moving means. The method of claim 1, And said movable stage further comprises a temperature control means capable of adjusting the temperature of said table. delete The method of claim 1, The irradiation means is a small animal fluorescent image simultaneous observation optical molecular imaging device, characterized in that it comprises only a LED light source or a white light source. The method of claim 1, When a plurality of specific positions are selected from the images taken by the whole monitoring photographing means, the stages are stored in the selected order and the specific positions of the subjects are located in a position where they can be photographed by the enlarged precision photographing means in the stored order. Small animal fluorescent imaging simultaneous observation optical molecular imaging device, characterized in that the movement means for controlling the table on which the subject is located.

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