This application claims priority from Korean Patent Application No. 10-2011-0055819, filed on Jun. 9, 2011, in the Korean Intellectual Property Office, the disclosure of which is incorporated herein in its entirety by reference.
1. Field
The present invention relates to a fluorescence detector.
2. Description of the Related Art
In amplifying nucleic acid, devices to check whether nucleic acid amplification is effectively performed after every cycle are required. In general, these devices detect fluorescence emitted from a sample to determine whether the nucleic acid amplification is effectively performed. That is, when a complementary probe is attached to a nucleic acid, and then, an excitation light is applied thereto, the excited probe emits fluorescence, and the devices detect the fluorescence.
However, the devices to detect fluorescence according to the related art are huge in size and heavy in weight, and when fluorescence or an excitation light passes through an optical system, a wavelength shift occurs such that interference occurs between several lights, and thus, a detection accuracy deteriorates.
One or more exemplary embodiments provide a fluorescence detector that detects fluorescence from a sample.
According to an aspect of an exemplary embodiment, there is provided a fluorescence detector for detecting fluorescence emitted from at least one sample contained in at least one sample unit, the fluorescence detector including at least one irradiating module which irradiates an excitation light to the sample; a fluorescence selecting unit which selectively transmits fluorescence emitted from the sample; a light-receiving unit which detects the fluorescence; and a telecentric lens positioned between the fluorescence selecting unit and the light-receiving unit, wherein each of the at least one irradiating module comprises: at least one light source which emits a light; an excitation light selecting unit which converts the light emitted from the at least one light source into the excitation light; and a beamsplitter which controls the excitation light to travel to the sample, and transmits the fluorescence emitted from the sample.
The irradiating module may further include a collimation lens which is disposed between the at least one light source and the excitation light selecting unit, and collimates the light emitted from the at least one light source.
The irradiating module may be movable in a first direction.
The number of the at least one irradiating module is two or more, and the two or more irradiating modules are disposed in parallel along one direction, and wherein respective excitation light selecting units of the two or more irradiating modules convert respective lights emitted from respective at least one light source into respective excitation lights having different wavelengths.
The at least one light source may include at least one light-emitting diode (LED) lighting.
The number of the at least LED lighting may be equal to the number of the at least one sample unit.
The fluorescence selecting unit may include an emission filter.
The emission filter may be formed of a plurality of the emission filters, and the plurality of emission filters may be changeable with respect to one another.
The light-receiving unit may include a charge-coupled device (CCD) camera.
The beamsplitter may include a dichroic filter.
An incident angle of the excitation light that is incident on the dichroic filter may be between 40 degrees and 50 degrees.
The excitation light selecting unit may include an excitation filter.
An incident angle of light from the irradiating module that is incident on the excitation filter may be between −5 degrees and 5 degrees.
The excitation light is a selected wavelength light.
The irradiating module may further include a wave guide which is disposed between the at least one light source and the excitation light selecting unit and divides the light emitted from the at least one light source into a plurality of lights that are parallel to one another.
The wave guide may be a glass wave guide which is integrally formed of a glass material.
The wave guide may include a plurality of optical fibers, and side ends of the plurality of optical fibers toward the at least one light source contact one another, and side ends of the plurality of optical fibers toward the excitation light selecting unit are separate from one another.
According to an aspect of another exemplary embodiment, there is provided a fluorescence detector for detecting fluorescence emitted from at least one sample contained in at least one sample unit, the fluorescence detector including an irradiating module which irradiates an excitation light to the sample; a fluorescence selecting unit which selectively transmits fluorescence emitted from the sample; a light-receiving unit which detects the fluorescence; and a telecentric lens positioned between the fluorescence selecting unit and the light-receiving unit, wherein the irradiating module includes: a light source comprising at least one laser diode which emits a light; and a beamsplitter which controls the light emitted from the at least one laser diode of the light source to travel to the sample, and transmits the fluorescence emitted from the sample.
A wavelength of the light emitted from the at least one laser diode may be materially the same as a wavelength of light capable of exciting a fluorescent material comprised in the sample.
The irradiating module may further include an excitation light selecting unit which is disposed between the light source and the beamsplitter and converts the light emitted from the light source into the excitation light.
The above and other aspects will become more apparent by describing in detail exemplary embodiments thereof with reference to the attached drawings, in which:
Hereinafter, the inventive concept will be described in detail by explaining exemplary embodiments with reference to the attached drawings.
The fluorescence detector 100 may include an irradiating module 110, a fluorescence selecting unit 120, a telecentric lens 130, and a light-receiving unit 140, and may further include a housing 150 that accommodates at least some of the aforementioned elements.
The fluorescence detector 100 is a device which determines an existence or an amount of a target included in the sample T by exciting the target and then detecting the fluorescence (FL) emitted from the target.
In the present embodiment, the target included in the sample T is a nucleic acid, and a probe complementary to a particular nucleic acid is attached to the particular nucleic acid, and then the excitation light EL is irradiated thereto so as to excite the probe. Afterward, by detecting the fluorescence emitted from the excited probe, it is possible to determine an existence or an amount of the probe, and based on the determination, an existence or an amount of the particular nucleic acid may be determined. This procedure will be described below in detail.
In general, the fluorescence detector 100 may be used in a polymerase chain reaction (PCR). That is, the fluorescence detector 100 may monitor an amplified amount of a nucleic acid after the PCR is completed at every cycle.
The sample T is included in a sample group 160. The sample group 160 includes one or more sample units 161, and the sample group 160 of
The irradiating module 110 irradiates the excitation light EL to the sample T, and includes a light source 111, a collimation lens 112, an excitation light selecting unit 113, and a beamsplitter 114.
The irradiating module 110 may be positioned above the sample group 160 and may move along a first direction D1. Also, the irradiating module 110 may include a plurality of the irradiating modules 110, and the plurality of irradiating modules 110 may be disposed in parallel to each other along one direction (in the present embodiment, they are disposed along the first direction D0. Here, each of the irradiating modules 110 may be regarded as a channel, and the irradiating modules 110 of the channels may be used to detect different types of nucleic acids. To do so, the irradiating modules 110 of the channels may emit lights having different wavelengths so as to excite probes to be attached to the different types of nucleic acids, and to do so, the irradiating modules 110 of the channels may include different types of light sources 111 or different types of excitation light selecting units 113. In the irradiating module 110 of
The light source 111 includes one or more light-emitting diode (LED) lightings. In order to observe various samples T, the LED lightings may include various types of LEDs according to wavelengths required to excite the various samples T. In the present embodiment, the LED lightings may have wavelengths of 470 nm, 520 nm, 590 nm, 640 nm, or the like but one or more embodiments are not limited thereto. Alighting with an appropriate wavelength may be used according to a target sample T, and the lighting is not limited to LED lighting.
The number of LED lightings included in the light source 111 may match with the number of sample units 161 to be observed. Referring to
The collimation lens 112 is positioned in a rear side of the light source 111 on an optical path, and the collimation lenses 112 are arranged to correspond to the LED lightings included in the light source 111, respectively. The collimation lens 112 collimates a travel path of light emitted from the light source 111.
The excitation light selecting unit 113 may be positioned in a rear side of the collimation lens 112 on the optical path and may include an excitation filter. The excitation light selecting unit 113 receives the light emitted from the collimation lens 112 and then outputs the excitation light EL that has a particular wavelength so as to excite the sample T. In more detail, the light emitted from the LED lighting has a Gaussian distribution with respect to a wavelength, and in this regard, when the light passes through the excitation filter, other portions of the light except for a portion of the light with a particular band wavelength do not pass through the excitation filter, and only the portion of the light with the particular band wavelength passes through the excitation filter.
In the present embodiment, the excitation light selecting unit 113 has one excitation filter with respect to lights emitted from the three LED lightings, but one or more embodiments are not limited thereto and the excitation light selecting unit 113 may include excitation filters that correspond to LED lightings, respectively.
The light that collimates after passing through the collimation lens 112 is incident on the excitation filter, and in this regard, an incident angle of the light with respect to the excitation filter may be between about −5 degrees and about 5 degrees, and it is optimal when the incident angle is 0 degrees. With the aforementioned range of incident angle, a wavelength shift that may occur when the light passes through the excitation filter may be effectively prevented and a wavelength of the light passing through the excitation filter may be accurately controlled.
The beamsplitter 114 is positioned in a rear side of the excitation light selecting unit 113 on the optical path, and functions to reflect the excitation light EL and to transmit fluorescence. The beamsplitter 114 may include a dichroic filter. The dichroic filter has a property of transmitting a light having a particular wavelength and of reflecting lights having the rest of the wavelengths. The dichroic filter used in the present embodiment may transmit a light having a wavelength of fluorescence emitted from the sample T and may reflect lights having the rest of the wavelengths including a wavelength of the excitation light EL.
The excitation light EL emitted from the excitation light selecting unit 113 is incident on the beamsplitter 114. Here, an incident angle of the excitation light EL with respect to the beamsplitter 114 may be between about 40 degrees and about 50 degrees, and it is optimal when the incident angle is 45 degrees. When a value of the incident angle is limited to the aforementioned range, a wavelength shift that may occur when a light is reflected by the dichroic filter may be effectively reduced, so that the wavelengths may be accurately controlled and detection of the fluorescence may be further accurately performed.
Moving units 115 are formed on a top surface of an irradiating module cover 116 and move the irradiating module 110 in the first direction D1. In the present embodiment, the first direction D1 is parallel to a direction in which 12 sample units 161 are arrayed in a single row in the sample group 160. However, one or more embodiments are not limited thereto and thus the first direction D1 may be changed according to a designer's objective.
The moving unit 115 includes a rail arranged on the irradiating module 110 and a rail arranged at a lower part of the housing 150. Both the rails engage with each other and are movable relatively to the other. The movement of the rails may be driven by a motor (not shown). However, one or more embodiments are not limited thereto, and thus, a way that the moving units 115 move the irradiating module 110 in one direction may be other ways for moving objects according to the related art.
A fluorescence selecting unit 120 is a portion to which the fluorescence emitted from the sample T is incident after passing through the beamsplitter 114. The fluorescence selecting unit 120 may include an emission filter.
The fluorescence selecting unit 120 transmits only a light having a wavelength of the fluorescence among incident lights. That is, a light emitted from the LED lightings is highly intensive, compared to the fluorescence emitted from the sample T. Thus, if the light from the LED lighting is directly incident to the light-receiving unit 140 for detection, a detection accuracy result considerably deteriorates. The fluorescence selecting unit 120 may be arranged to prevent the deterioration and may be formed in a path on which a light emitted from the sample T travels to the light-receiving unit 140. In the present embodiment, the fluorescence selecting unit 120 is formed in the irradiating module 110. However, one or more embodiments are not limited thereto, and thus, the fluorescence selecting unit 120 may be positioned in the housing 150.
Referring to the modified example of
In order to transmit fluorescence lights having different wavelengths which occur via a plurality of channels of the irradiating module 110, the change of the emission filters is performed by the filter changing unit 120a.
For example, when an excitation light EL having a first excitation light wavelength is irradiated to the sample T via a first channel, if it is assumed that fluorescence having a first fluorescence wavelength is emitted from the sample T, a first emission filter for transmitting the fluorescence having the first fluorescence wavelength is mounted by the filter changing unit 120a. Afterward, when an excitation light EL having a second excitation light wavelength is irradiated to the sample T via a second channel, if fluorescence having a second fluorescence wavelength is emitted from the sample T, a second emission filter that corresponds to the fluorescence having the second fluorescence wavelength is mounted by the filter changing unit 120a.
Thus, in correspondence to fluorescence lights having different wavelengths which occur via the channels of the irradiating module 110, the filter changing unit 120a selectively mounts the emission filter for transmitting a corresponding fluorescence light in the fluorescence selecting unit 120. That is, by selectively choosing the emission filter that corresponds to one of the different wavelengths, the fluorescence lights having the different wavelengths may be detected.
The telecentric lens 130 is disposed on a path on which fluorescence emitted from the sample T travels to the light-receiving unit 140, so that the fluorescence passes through the fluorescence selecting unit 120 and then passes through the telecentric lens 130. The telecentric lens 130 may include various optical elements and may be formed by combining the various optical elements. In the present embodiment, the telecentric lens 130 includes only one lens but one or more embodiments are not limited thereto.
The telecentric lens 130 transmits only light with a small allowance incident angle (e.g., between about −2 degrees and about 2 degrees) from among lights that are incident on the telecentric lens 130. That is, when lights are incident on the telecentric lens 130 via various paths, only a light that is approximately parallel to a particular direction passes through the telecentric lens 130. Because the telecentric lens 130 has the small allowance incident angle, when light passes through the telecentric lens 130, a wavelength shift does not occur. Thus, it is possible to accurately control a wavelength of fluorescence, and interference between fluorescence lights of different wavelengths generated via the multiple channels does not occur. Therefore, an accurate detection result may be achieved.
The light-receiving unit 140 may detect fluorescence emitted from the sample T and may include various types of detection sensors. In the present embodiment, the light-receiving unit 140 includes a charge-coupled device (CCD) camera 141 as a sensor for detecting the fluorescence (FL).
Hereinafter, an operation process of the fluorescence detector 100 will be described.
Referring to
First, as illustrated in
Hereinafter, the fluorescence FL detection process will be described with reference to
As the sample T is excited, the sample T emits fluorescence. Here, the fluorescence moves along a third direction D3 and keeps moving after passing through the dichroic filter of the beamsplitter 114. Afterward, the fluorescence passes through the fluorescence selecting unit 120 positioned in the irradiating module 110, and in this process, the excitation light EL of the light incident on the fluorescence selecting unit 120 is filtered, and only the fluorescence passes through the fluorescence selecting unit 120. Afterward, when the fluorescence passes through the telecentric lens 130, only the fluorescence of which an incident angle with respect to the telecentric lens 130 is between about −2 degrees and about 2 degrees passes through the telecentric lens 130, and after this process, the fluorescence becomes parallel, is incident on the light-receiving unit 140 disposed in a top portion, and then is detected.
The irradiating module 110 of
For example, after the first channel of the irradiating module 110a irradiates the excitation light EL to a first sample unit 160a that is first positioned in the first direction (D1), the first channel of the irradiating module 110a moves in the first direction (D1) and then irradiates the excitation light EL to a second sample unit 160b. Afterward, the second channel of the irradiating module 110b slightly moves in the first direction (D1), and irradiates the excitation light EL to the second sample unit 160b. Afterward, the four channels of the irradiating modules 110a, 110b, 110c, and 110d move in the first direction (D1) and sequentially perform the aforementioned processes.
In this regard, when the four channels of the irradiating module 110 irradiate the excitation light EL to each sample unit, an emission filter that corresponds to a wavelength of fluorescence (FL) emitted from each sample T may be changed by the filter changing unit 120a.
In the present embodiment, the irradiating module 110 includes a plurality of the light sources 111 and a plurality of the collimation lenses 112, but a structure of the irradiating module 110 may vary.
Referring to
The light source 111A of the irradiating module 110A of
The wave guide 112A is disposed between the light source 111A and the excitation light selecting unit 113, and divides a light from the light source 111A into a plurality of lights that are parallel to each other. The wave guide 112A may be a glass wave guide that is integrally formed of a glass material. The wave guide 112A of the irradiating module 110A of
Thus, by using the irradiating module 110A of
In addition, the wave guide 112A of the irradiating module 110A of
Referring to
Also, a light source of an irradiating module may include a laser diode.
Referring to
The plurality of laser diodes of the light source 111C are vertically disposed and irradiate a laser toward the beamsplitter 114. The laser diode emits the laser formed of light having a narrow frequency band. Because the laser diode is well known, descriptions of its structure and operational principle are omitted here. Because the laser has an excellent straight-advance characteristic and does not disperse, when the light source 111C has the plurality of laser diodes, a collimation lens is not separately required between the light source 111C and the beamsplitter 114. Thus, an increase in manufacturing costs due to an installation of the collimation lens may be effectively restricted, and the irradiating module 110C may have a simple structure. The laser diode of the light source 111C may emit the laser having materially the same frequency band as an excitation light capable of exciting a fluorescent material of a sample. The excitation light selecting unit 113 may be disposed between the plurality of laser diodes of the light source 111C and the beamsplitter 114 to block a light having another wavelength other than a wavelength corresponding to the excitation light. However, when the plurality of laser diodes of the light source 111C have materially the same frequency band as the excitation light, the excitation light selecting unit 113 may not be separately disposed between the plurality of laser diodes and the beamsplitter 114.
The one or more embodiments may provide a fluorescence detector having an excellent fluorescence detection performance.
While the inventive concept has been particularly shown and described with reference to exemplary embodiments thereof, it will be understood by those of ordinary skill in the art that various changes in form and details may be made therein without departing from the spirit and scope of the inventive concept as defined by the following claims.
Number | Date | Country | Kind |
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10-2011-0055819 | Jun 2011 | KR | national |