KR101821637B1 - Luminescence microscope - Google Patents

Abstract

The present invention relates to a luminescence microscope capable of detecting fluorescence of an observation sample by irradiating a plurality of excitation lights with a plurality of excitation light sources, comprising: an LED light source module (100) composed of at least more than two LEDs (110) which generate excitation lights of specific wavelengths different from each other; a detecting unit (200) for detecting emitted light generated in the sample; an optical filter module (300) including two or more optical filters (310) provided at a front end of the detecting unit (200) and capable of blocking the excitation light among lights incident on the detecting unit (200) and corresponding to the LED (110); and one beam splitter (400) in which the excitation light generated from the LED (110) is directly irradiated and reflected in the direction of a sample (2), and in which the emitted light generated in the sample (2) is transmitted.

Description

Luminescence microscope

The present invention relates to a light emitting microscope having a plurality of excitation light sources and capable of detecting light emission of an observation sample by irradiation of a plurality of excitation lights.

The electrons in the material can be transited from the ground state to the excited state by various stimuli. The electrons in this excited state return to the ground state in a stable state again, Which is a phenomenon that emits light corresponding to the light. The luminescence may be a photoexcitation luminescence, an electron excitation luminescence, a cathode luminescence, a fluorescence, a phosphorescence or the like depending on excitation sources here.

Among them, fluorescence microscopy is a technique in which a fluorescent substance (fluorescent pigment) is treated on a sample using the principle that a fluorescent substance emits fluorescence when the light absorbs a specific wavelength of light, and then the sample is irradiated with light having an absorption wavelength of the fluorescent substance, Is a device that observes a sample through the emitted radiation. Fluorescence microscopy is widely used when observing a sample such as a biochip because a clear image can be obtained compared with a general optical microscope.

1 is a schematic view showing a general fluorescence microscope.

1, the fluorescence microscope comprises a light source 10, a first optical filter 21 for selectively transmitting monochromatic light of a specific wavelength among the light emitted from the light source 10 and transmitting exciting light, An objective lens 22 for irradiating excitation light to the sample 2 placed on the plate 1, a detector 30 for detecting emitted light generated from the sample 2, a detector 30 A second optical filter 23 provided at the front end for blocking the excitation light from the light incident on the detector 30 and a second optical filter 23 disposed on the optical path between the objective lens 22 and the detector 30, And a dichroic mirror (DM) 24 through which the excitation light having passed through the objective lens 21 is reflected by the objective lens 22 and the light emission signal generated from the sample 2 is transmitted.

The first optical filter 21 (hereinafter also referred to as an 'excitation light filter') is used as a continuous light source (or a white light source) in a light source unit 10 Only the monochromatic light (excitation light) coinciding with the absorption wavelength of the phosphor attached to the sample 2 is selectively transmitted through the white light source generated in the sample 2 to be irradiated onto the sample 2.

The dichroic mirror 24 reflects the excitation light transmitted through the first optical filter 21 in the direction of the sample 2 and the emitted light generated in the sample 2 is reflected by the dichroic mirror 24 And is detected by the detector 30.

On the other hand, the ideal dichroic mirror 24 reflects all of the incident excitation light, and all of the emitted light generated from the sample 2 must be transmitted. However, since the excitation light is not completely blocked, The excitation light is cut off by the two-light filter 23.

In general, the wavelength lambda 1 of the emitted light in the sample is longer than the wavelength lambda 2 of the excitation light (lambda 2 <lambda 1), and thus the long-pass filter LP is used for the second optical filter.

Such a fluorescence microscope is used to observe the image of a component having a luminescence property in an observation sample with high resolution, and is widely used in biological, medical, and semiconductor fields.

However, in a conventional fluorescence microscope, a mercury (Hg) lamp or a xenon (Xe) lamp used as a light source for exciting a sample is very harmful to the environment and its use is gradually restricted internationally, It has the possibility that its use can be prohibited.

Such a light source uses a characteristic of spectra emitted from a specific atomic species due to a discharge in a gaseous state at a high pressure. It is difficult to maintain a constant state when used for a long time in the manufacturing principle, and a very short lifetime And there is an inconvenience in maintenance and management due to frequent replacement.

In the conventional fluorescence microscope, only the monochromatic light of a specific wavelength is selected by using the excitation light filter among the light generated from the continuous light source, and excitation light not completely removed from the DM is removed The LP filter is indispensable, and therefore, the cube having the combination of the three optical components consisting of the excitation light filter, the dichroic mirror, and the long wavelength transmission filter must be used according to the number of the light sources to be used.

In particular, it is necessary to observe the precise shape of the sample 2 by obtaining a fluorescence image by irradiating various light to compare them, and a multi-fluorescence microscope including a plurality of light sources capable of irradiating various excitation light to the sample Has been developed.

In the present multi-fluorescence microscope, a combination of two or more cube is sold in combination of two to eight. If the wavelength of the light source is increased, the number of the cube optical system is increased, .

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Japanese Patent Application Laid-Open No. 10-2009-0118774 (published on November 18, 2009) Published Patent Publication No. 10-2010-0087141 (Published Date: Aug. 03, 2010)

SUMMARY OF THE INVENTION It is an object of the present invention to provide a light emitting microscope for detecting light emission of a sample using a plurality of excitation light sources, It is intended to provide a microscope.

According to an aspect of the present invention, there is provided a light emitting microscope comprising: an LED light source module including at least two LEDs for generating excitation light having different wavelengths; A detector for detecting emitted light generated in the sample; An optical filter module provided at a front end of the detection unit and including at least two optical filters for shielding excitation light from light incident on the detection unit and corresponding to the LEDs and blocking excitation light; And a beam splitter through which the excitation light generated from the LED is directly irradiated and reflected in the sample direction and the emitted light generated in the sample is transmitted.

Preferably, the optical filter module includes: a fixing member having the optical filter arranged and fixed in a plane; And a driving unit for driving the fixing member. More preferably, the fixing member includes a rotation axis rotatable by the driving unit.

Preferably, the LED light source module comprises: a fixing member in which the LED is arranged and fixed in a plane; And a driving unit for driving the fixing member.

Preferably, the beam splitter has a reflectance / transmittance ratio of 0.5 or less.

The light emitting microscope according to the present invention comprises an LED light source module constituted by a plurality of LEDs which are monochromatic lights of high output power as an excitation light source and an optical filter module provided at the front end of the detection section and composed of a plurality of optical filters corresponding to the LEDs, And a beam splitter through which the excitation light generated from the LED is directly irradiated and reflected in the sample direction, and the emitted light generated in the sample is transmitted, thereby eliminating a separate excitation light filter for selecting specific excitation light And the fluorescence image of the sample can be detected with only one beam splitter regardless of the wavelength of the excitation light source, thereby simplifying the configuration of the apparatus and facilitating maintenance.

1 is a schematic view showing a general fluorescence microscope,
2 is a configuration diagram of a light emitting microscope according to the present invention,
3 (a) and 3 (b) are a configuration diagram of an LED light source module according to another embodiment,
4 is a planar view of the optical filter module in the light emitting microscope of the present invention.

The specific structure or functional description presented in the embodiment of the present invention is merely illustrative for the purpose of illustrating an embodiment according to the concept of the present invention, and embodiments according to the concept of the present invention can be implemented in various forms. And should not be construed as limited to the embodiments described herein, but should be understood to include all modifications, equivalents, and alternatives falling within the spirit and scope of the invention.

Meanwhile, in the present invention, the terms first and / or second etc. may be used to describe various components, but the components are not limited to the terms. The terms may be referred to as a second element only for the purpose of distinguishing one element from another, for example, to the extent that it does not depart from the scope of the invention in accordance with the concept of the present invention, Similarly, the second component may also be referred to as the first component.

Whenever an element is referred to as being "connected" or "connected" to another element, it may be directly connected or connected to the other element, but it should be understood that other elements may be present in between something to do. On the other hand, when it is mentioned that an element is "directly connected" or "directly contacted" to another element, it should be understood that there are no other elements in between. Other expressions for describing the relationship between components, such as "between" and "between" or "adjacent to" and "directly adjacent to" should also be interpreted.

The terminology used herein is for the purpose of describing particular embodiments only and is not intended to be limiting of the invention. The singular expressions include plural expressions unless the context clearly dictates otherwise. It will be further understood that the terms " comprises &quot;, or "having &quot;, and the like in the specification are intended to specify the presence of stated features, integers, But do not preclude the presence or addition of steps, operations, elements, parts, or combinations thereof.

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

2, the light emitting microscope of the present invention includes an LED light source module 100 including a plurality of LEDs 110, a detecting unit 200 for detecting emitted light generated from the sample 2, An optical filter module 300 including a plurality of optical filters 310 and an optical filter 300 which reflects the excitation light generated from the LED 110 in the direction of the sample and transmits the emission light emitted from the sample in the direction of the detection unit 200 And one beam splitter (400).

In the present invention, a light emitting diode (LED) 110, which is a monochromatic light, is used as an excitation light source, and may be composed of a plurality of LEDs to generate excitation light having a specific wavelength.

Meanwhile, the LED light source module may be an LED that emits blue, green, red, yellow, or light of a specific wavelength, and may further include a white LED. have.

Unlike mercury lamps or xenon lamps, LEDs have a long lifespan, so maintenance is easy in accordance with the replacement cycle, and there is little drop in light quantity according to lighting time, so that a stable image can be obtained. In addition, the LED can easily realize light output of several watts (W) according to the input current.

In particular, a mercury lamp or a xenon lamp requires an excitation light filter to select only the excitation light. In this case, since the excitation light transmitted through the excitation light filter is insufficient, a dichroic mirror is required to concentrate the light with high reflectance, The present invention is characterized in that a separate excitation light filter is not used by using an LED which is a monochromatic light with high output.

Preferably, the LED light source module 100 includes a first fixing member 120 in which the LEDs 110 are arranged and fixed in a planar manner, a first driving unit 130 for driving the first fixing member 120; . &Lt; / RTI &gt;

The first fixing member 120 may include a plurality of LEDs 110 fixed and rotatable and connected to the first driving unit 130 by a first rotating shaft 131 so as to be coupled to the first driving unit 130, The specific LED 110 among the plurality of LEDs 110 may be selected and used as an excitation light source by the rotational driving of the LEDs 110. [ Each of the LEDs 110 is connected to a power source (not shown) so that lighting control can be performed.

3A and 3B are block diagrams of an LED light source module according to another embodiment.

3 (a) and 3 (b), the LED light source module 600 of the present embodiment includes a plurality of LEDs 610 and 621 for generating excitation light of different wavelengths, The LEDs 610 and 621 may include a first LED 610 and a plurality of second LEDs 621 arranged to have an optical axis orthogonal to the first LED 610.

Each of the second LEDs 621 is provided with a reflection mirror 622 that totally reflects light in the optical axis direction of the first LED 610 and the reflection mirror 622 is provided with a drive arm 623 connected to a drive unit It is possible to move forward and backward.

For example, each reflective mirror 622 may be turned off (off) on the optical axis C of the first LED 610. For example, each reflective mirror 622 may be turned off on the optical axis C of the first LED 610, The first LED 610 emits light and excitation light is generated in the first LED 610 or the specific reflection mirror 622 is positioned on the optical axis C so that the corresponding second LED 621 emits light So that excitation light can be generated.

Referring again to FIG. 2, the detection unit 200 is for detecting emitted light of the sample 2, and can be provided by a well-known CCD image element or a CMOS image element to obtain a fluorescent image of the sample.

The optical filter module 300 is disposed at the front end of the detection unit 200 to block the excitation light from the LED 110 in the incident light of the detection unit 200. Preferably, And at least two optical filters (310) capable of blocking light.

The optical filter module 300 includes a second fixing member 320 in which the optical filter 310 is arranged and fixed in a plane and a second driving unit 330 for driving the second fixing member 320 can do.

The second fixing member 320 is connected to the second driving unit 330 by the second rotating shaft 331 as a rotatable member that is fixed and rotatable by the plurality of optical filters 310 so that the second driving unit 330 The specific optical filter 310 among the plurality of optical filters 310 can be selected to block excitation light.

FIG. 4 is a plan view of the optical filter module according to the present invention. The optical filter module may include a plurality of optical filters capable of blocking excitation light according to the configuration of the LED, (B), green (G), red (R), yellow (Y), or a specific wavelength (?) Corresponding to the LED light source by a Stokes shift Can be provided by a long-wavelength transmission filter in which filtering is performed on the signal. On the other hand, the second fixing member 320 may further include a window through which the white light W can be transmitted, and this window may be used for reflection image observation when a white LED is used as the excitation light.

2, the beam splitter 400 is positioned at the intersection of the LED light source module 100 and the detecting unit 200 arranged at right angles to each other, and excitation light generated from the LED 110 is directed toward the sample 2 And the emitted light generated in the sample 2 is transmitted in the direction of the detection unit 200.

Particularly, in the present invention, the beam splitter 400 is provided as one with a certain reflectance / transmittance ratio and is characterized in that the excitation light is reflected by the sample to a plurality of LED light sources and a plurality of optical filters, and the emitted light of the sample is transmitted do.

Preferably, in the present invention, the beam splitter 400 has a reflectance / transmittance ratio of 0.5 or less, thereby lowering the reflectance of the excitation light generated from the LED 110 and increasing the transmittance of the emitted light generated from the sample 2 The amount of emitted light of the sample incident on the detection unit 200 can be increased. As described above, since sufficient light output can be obtained in accordance with the applied current, the LED for monochromatic light can transmit sufficient excitation light to the sample 2 even if the reflectance of the excitation light is low in the beam splitter 400, The beam splitter 400 preferably has a reflectance / transmittance ratio of 0.5 or less.

Reference numeral 500 denotes an optical mechanism for collecting and irradiating excitation light reflected by the beam splitter 400 onto a specimen 2 and guiding emission light emitted from the specimen 2 to the detection unit 200, It is to be understood that the present invention can be also applied to an optical path incident on the detector 200 as needed.

In the light emitting microscope of the present invention configured as described above, the specific LED 110 selected from the LED light source module 100 is used as the excitation light according to the sample 2 to be irradiated. At this time, in the optical filter module 300, The fluorescence image of the sample can be obtained by detecting the emitted light generated from the sample 2 in the detection unit 200. In this case,

Particularly, in the present invention, it is possible to exclude a separate excitation light filter for selecting specific excitation light by using an LED, which is monochromatic light with high output power as an excitation light source, and to use only one beam splitter 400 It is possible to detect fluorescence image of sample and simplify equipment configuration and maintenance is convenient.

It will be apparent to those skilled in the art that various modifications and variations can be made in the present invention without departing from the spirit or scope of the inventions. It will be apparent to those of ordinary skill in the art.

100: LED light source module 110: LED
120: first fixing member 130: first driving part
200: Detector 300: Optical filter module
310: optical filter 320: second fixing member
330: second driving part 400: beam splitter

Claims (6)

An LED light source module (100) including at least two LEDs (110) generating excitation light of different wavelengths;
A detector (200) for detecting emitted light generated in the sample;
Two or more optical filters 310 provided at the front end of the detecting unit 200 to block the excitation light from the light incident on the detecting unit 200 and to block excitation light corresponding to the LEDs 110 An optical filter module 300;
And a beam splitter (400) having a reflectance / transmittance of 0.5 or less, wherein the excitation light generated from the LED (110) is directly irradiated and reflected in the sample direction, and the emitted light generated in the sample is transmitted,
The LED light source module (100)
A first LED 610;
A plurality of second LEDs 621 arranged to have an optical axis orthogonal to an optical axis C of the first LED 610;
A plurality of reflection mirrors 622 provided to each of the second LEDs 621 so as to totally reflect light in the direction of the optical axis C;
And a first driving unit that selectively drives the driving arm 623 provided in each of the reflection mirrors 622 in a direction perpendicular to the optical axis C,
The optical filter module (300)
A fixing member 320 rotatable with the rotation axis 331, the optical filter 310 being arranged and fixed in a plane;
And a second driving part (330) coupled to the rotation axis (331) of the fixing member (320) to rotationally drive the fixing member (320). delete delete delete delete delete

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