JP2006258776A - Particle classifier - Google Patents

Abstract

<P>PROBLEM TO BE SOLVED: To provide a particle classifier capable of attaining properly and efficiently analysis as to fluorescence of a measuring objective particle. <P>SOLUTION: In this particle classifier for detecting scattered light and the fluorescence generated from the measuring objective particle 15, and for analyzing the measuring objective particle, based on detected signals detected therein, by irradiating a flow of a solution containing the the measuring objective particle 15 such as a cell, a chromosome, a biopolymer contained therein, and the like in a flow cell 14, with laser beams from laser beam sources 10a, a light emitting diode (LED) 30 capable of emitting a blue range of light beam as an excitation beam is provided as the light source for fluorescence excitation for the measuring objective particle 15, and fluorescence detectors are provided to obtain respective required fluorescence signals, as to the light beams of the scattered light and the fluorescence excited by irradiating the measuring objective particle with the respective light beam sources, via a filter 32 for removing a scattered light wavelength. <P>COPYRIGHT: (C)2006,JPO&NCIPI

Description

  The present invention irradiates a flow of an aqueous solution containing cells, chromosomes and biopolymers contained therein with laser light, detects fluorescence and scattered light emitted from the cells, etc. The present invention relates to a particle classification device that performs counting.

  The particle classification device is (1) as a means for measuring the relative amount of intracellular DNA, RNA, enzyme, protein, etc., (2) as a means for examining functions such as cell activity, antibody productivity, enzyme activity, and (3) Widely used as a means of automatically classifying cell, chromosome, lymphocyte and other types.

  Conventionally, as this kind of particle classification device, for example, a blood sample is diluted and stained to obtain a sample liquid, and this sample liquid is made to flow in the center of the flow cell in a trickle, and the narrowly focused light from the light source is applied to the trickle part. A detection unit is formed by irradiation, and a scattered light and a change in fluorescence generated each time a blood cell passes through the detection unit one by one are detected by a photodetector, and the scattered light intensity and fluorescence are detected from the detected signal. A configuration is known in which a two-dimensional distribution map with two axes of intensity is created, and a classification line is set in the two-dimensional distribution map, thereby classifying and counting each particle.

  The applicant has previously developed a particle classification device that can classify white blood cells containing small granules or particles by detecting forward scattered light, side scattered light and back scattered light scattered from white blood cells. As a result, we succeeded in developing an apparatus that can accurately classify cells with a relatively low content, and obtained a patent (see Patent Document 1).

  The particle classification apparatus in Patent Document 1 is (1) a particle classification apparatus that classifies particles by irradiating light to the particles and detects scattered light from the particles, and (2) detects forward small angle scattered light. Forward small angle scattered light detection means, and (3) forward large angle scattered light detection means that is disposed between the forward small angle scattered light detection means and the side scattered light detection means and detects forward large angle scattered light, and (4a) The detection signals corresponding to the forward small angle scattered light, the side scattered light and the forward large angle scattered light output from the forward small angle, forward large angle and side scattered light detection means are input to calculate the respective scattered light intensity data. (4b) Two-dimensional distribution data of the forward small angle scattered light intensity and the forward large angle scattered light intensity from the forward small angle, forward large angle and side scattered light intensity data, and the forward small angle scattered light intensity and the side scattered light. A hand to calculate 2D intensity distribution data And (4c) an analysis means comprising: a means for determining a specific white blood cell region from the two-dimensional distribution data of the forward small angle scattered light intensity and the forward large angle scattered light intensity; and (5) the calculated forward small angle scattering. Two-dimensional distribution data of light intensity and forward large-angle scattered light intensity, two-dimensional distribution data of forward small-angle scattered light intensity and side-scattered light intensity, and the above-mentioned specified white blood cell data are converted into the forward small-angle scattered light intensity and side-scattered data. And display means for displaying on the two-dimensional coordinates of the light intensity.

  Moreover, in the conventional particle classification apparatus, an argon laser is used as a light source for fluorescence excitation with respect to the measurement target particle. This is because in order to generate fluorescence in the measurement target particles, it is necessary to use light in a blue region having a relatively short wavelength as excitation light. However, the argon laser is expensive, and not only does the laser body itself occupy a large volume, but also the peripheral devices associated with the laser, such as the required power source for driving the laser, are large. In addition, there is a problem that overall power consumption is large.

  From such a viewpoint, conventionally, an optical particle analyzer using both a laser light source and a lamp light source as an irradiation light source has been proposed (see Patent Document 2). This optical particle analyzer uses lamp light for the excitation of sample particle fluorescence, and uses a semiconductor laser for scattered light, and the laser light and lamp light from these compact and low-cost light sources are in the particle trickle region of the flow cell. By being configured so as to cross and irradiate, scattered light and fluorescence of one particle can be detected at the same place and at the same timing. Also, since the laser light is narrowed down and has a single peak, the scattered light signal also becomes a peak signal, and the intensity distribution of the lamp light is detected by detecting the value of the fluorescent signal using this scattered light signal as a timing signal. Even if there is unevenness, a fluorescence signal correlated with the amount of fluorescence emitted from the particles can be detected.

  Furthermore, in the conventional optical particle analyzer, the laser light source is used for a detector that detects the intensity of the forward small-angle scattered light having a small scattering angle and a detector that detects the intensity of the forward large-small angle scattered light having a large scattering angle. As the optical system for receiving light, a collimator lens, a mirror with a hole, and a condensing lens are required, so the number of parts increases and the adjustment of the optical system becomes sophisticated and complicated. There is a problem that the area becomes large and it is difficult to reduce the size of the entire apparatus.

  In order to solve such problems, the present applicants can separate light of different scattering angles with a simple configuration, can easily adjust the optical system, have high light utilization, and further divide. Even if the number of areas increases, the number of parts of the optical system that separates the light does not increase, and the occupied area can be stopped only by increasing the space with the increase in the number of detectors without causing an increase in size. A compound lens that constitutes an optical system for receiving light to the detector, which can obtain a highly accurate particle analyzer, was successfully developed, and a patent application was filed (see Patent Document 3).

  That is, the particle analyzer described in Patent Document 3 (1) classifies the particles by separating and detecting the scattered light having different scattering angles from the scattered light generated by irradiating the particles with light. In the particle analyzer configured to be performed, (2) a plurality of ring-shaped lens elements having different focal positions arranged so as to block the optical path of the scattered light are integrated in a concentric circle state. A compound lens; and (3) a detector for detecting each scattering angle disposed at each of the scattered light imaging positions corresponding to the focal positions of the plurality of lens elements, and (4) detection by the plurality of detectors. Analyzing means for classifying the particles based on the output is provided.

Japanese Patent No. 3350775 JP-A-3-233344 Japanese Patent Laid-Open No. 11-23447

  However, conventionally, as a particle classification device capable of detecting scattered light and fluorescence and analyzing sample particles, one having the configuration shown in FIG. 3 is known. That is, in FIG. 3, reference numeral 10 b indicates a laser light source (scattered light excitation light source or fluorescence excitation light source), and laser light emitted from the laser light source 10 b passes through the light beam shaping lens 12 in the flow cell 14. Irradiated to the measurement target particle 15. In this way, the laser light applied to the measurement target particle 15 excites forward scattered light, side scattered light, and fluorescence, respectively. The laser light irradiated to the flow cell 14 is configured such that a direct light beam incident in front of the laser light is blocked by the light beam blocker 16.

  Thus, the forward scattered light excited about the measurement target particle 15 is collected by the scattered light condensing lens 18a, input to the forward scattered light detector 20a, and converted into an electrical signal. Further, the side scattered light and the fluorescence excited with respect to the measurement target particle 15 are collected by the scattered light and the fluorescent light collecting lens 19a and sequentially applied to the light beam splitters 22a, 22b, 22c, and 22d arranged in series. Incident.

  In the light beam splitter 22a, the side scattered light is reflected, and this reflected light beam is collected by the scattered light collecting lens 18b, input to the side scattered light detector 20b, and converted into an electric signal. On the other hand, the light beam transmitted through the light beam splitter 22a is subjected to the scattered light wavelength removal by the scattered light excitation light source or the scattered light wavelength removal from the fluorescence excitation light source via the scattered light wavelength removal filter 33, The light beams are sequentially incident on the light beam splitters 22b, 22c, and 22d. Then, the preset first fluorescence is reflected by the light beam splitter 22b, and this reflected light beam is condensed by the fluorescence condenser lens 19b via the wavelength selection filter 24a and input to the fluorescence detector 26a. And converted into an electrical signal. Similarly, the preset second fluorescence is reflected by the light beam splitter 22c, and this reflected light beam is condensed by the fluorescence condenser lens 19c via the wavelength selection filter 24b, and is applied to the fluorescence detector 26b. It is input and converted into an electrical signal. Further, the preset third fluorescence is reflected by the light beam splitter 22d, and this reflected light beam is condensed by the fluorescence condenser lens 19d via the wavelength selection filter 24c and input to the fluorescence detector 26c. Converted into an electrical signal. On the other hand, the transmitted light beam composed of the fourth fluorescence transmitted through the light beam splitter 22d is condensed by the fluorescence condenser lens 19e through the wavelength selection filter 24d, and input to the fluorescence detector 26d to be converted into an electrical signal. Is done.

  According to the particle classification apparatus configured as described above, the measurement target particles are classified into the fluorescence types as described above and measured, thereby analyzing the specificity of the various measurement target particles as described above. be able to.

  However, in the particle classification apparatus having the above configuration, when the measurement target particles are different, the type of fluorescence to be analyzed is different. For example, the setting of the wavelength selection filter is changed, and the sensitivity and threshold value of each detector are changed. The settings need to be optimized. The task of making such a change becomes extremely complicated. Therefore, in order to analyze many types of measurement target particles, the measurement target particles are specified in advance, and the sensitivity and threshold value of the detector for each wavelength selection filter are set. It is easy to install a plurality of optimized particle classification devices. However, since the particle classification device is expensive in terms of the light source and its peripheral devices, installing a particle classification device for each type of particle to be measured requires great equipment costs and installation space, and is economical. There are disadvantages.

  Therefore, the present inventor uses an argon laser as a light source for excitation of fluorescence with respect to a measurement target particle in the particle classification apparatus having the above-described configuration, and it is necessary to use light in a blue region having a relatively short wavelength as excitation light. Therefore, a light source consisting of a light emitting diode (LED) that can emit light in the blue region of a relatively short wavelength as excitation light is newly set as a light source for fluorescence excitation compared to conventional laser light sources. As a result, it has been found that a conventional laser light source can be used as a light source for exciting scattered light with respect to particles to be measured, so that a relatively low-cost apparatus configuration can be obtained. Moreover, in this case, it has been found that the analysis of the scattered light and fluorescence of the particles to be measured can be achieved more appropriately and efficiently.

  In the particle classification apparatus having the above configuration, since fluorescence is extremely weak light compared to scattered light, it may be important to increase sensitivity and S / N ratio. That is, direct light from the light source for fluorescence excitation and the light source for excitation of scattered light is reflected, refracted, and scattered by various optical elements and flow cells. Such light has an extremely large intensity compared to the fluorescence to be detected (this is called stray light). Such stray light enters the detector from other than the intended optical path. Therefore, in order to remove such stray light and increase the S / N ratio and sensitivity of fluorescence detection, a method of removing stray light using a pinhole is generally employed.

  Furthermore, it is important that the wavelength selection filter provided to obtain the fluorescence of the required selected wavelength is set so as to avoid oblique incidence since it is best when the light beam is incident perpendicularly. . In this case, the wavelength selective filter can be used with the wavelength selection performance in the best state by allowing the collimator to input parallel light to the wavelength selective filter.

  Then, as described in Patent Document 3 described above, the present inventor arranged a plurality of lens elements that are arranged so as to block the optical path of the scattered light and have different focal positions as concentric circles or the same circle. Using a combination of a compound lens integrated in a state and a plurality of detectors respectively arranged at the scattered light and fluorescence imaging positions according to the focal positions of the plurality of lens elements, It has been found that a particle classification device that can easily improve the sensitivity of fluorescence detection and the S / N ratio can be obtained by configuring to perform detection.

  Accordingly, an object of the present invention is to provide a light source for scattered light excitation and a light source for fluorescence excitation separately, and to scatter light and fluorescent light beams excited by irradiating each light source to the measurement target particle, A filter for removing the scattered light wavelength is provided, and each filter is configured to be incident on a fluorescence detector for obtaining a required fluorescence signal. In particular, the analysis of the fluorescence of the measurement target particle is performed more appropriately and efficiently. It is to provide a particle classification device that can be achieved.

In order to achieve the above object, a particle classification apparatus according to claim 1 of the present invention is a laser light source for a flow of an aqueous solution containing particles to be measured such as cells, chromosomes and biopolymers contained therein in a flow cell. In the particle classification apparatus for detecting the scattered light and fluorescence generated by the measurement target particle by irradiating the laser beam from and analyzing the measurement target particle based on these detected signals,
As a fluorescence excitation light source for the measurement target particles, a light emitting diode (LED) capable of emitting blue region light as excitation light is provided,
Fluorescence detectors for obtaining respective required fluorescence signals through a filter for removing scattered light wavelengths with respect to scattered light and fluorescent light beams excited by irradiating the respective light sources to the measurement target particles Is provided.

The particle classification apparatus according to claim 2 of the present invention is configured to irradiate the measurement target particle in the flow cell with the laser light emitted from the laser light source via the light beam shaping lens, and to detect the forward scattered light and the side light. Each of the scattered light is excited at the same time, and the light emitted from the fluorescence excitation light source is irradiated to the measurement target particle in the flow cell to excite the fluorescence,
The forward scattered light is configured to be collected by a scattered light collecting lens and input to a forward scattered light detector to be converted into an electrical signal,
The side scattered light and fluorescent light are collected by the scattered light and fluorescent light collecting lens, reflected through the light beam splitter, and further collected by the scattered light collecting lens and input to the side scattered light detector. A fluorescent condensing lens that converts the fluorescence obtained by passing through the light beam splitter and the light having a selected wavelength by a combination of a plurality of light beam splitters and wavelength selection filters. It is characterized in that the light is condensed by the above-mentioned and input to a fluorescence detector and converted into an electric signal.

  According to a third aspect of the present invention, there is provided the particle sorting apparatus according to the present invention, wherein the plurality of light beam splitters and wavelength selectors are used for a fluorescence detector that converts fluorescence obtained by transmitting the light beam splitter into an electrical signal. A filter for removing the wavelength of scattered light from the scattered light excitation light source is provided at the forefront of the combination of filters.

  According to a fourth aspect of the present invention, there is provided a particle classification apparatus including: a fluorescent light between the scattered light collecting lens and the forward scattered light detector, with respect to the forward scattered light detector that converts the forward scattered light into an electrical signal. A filter for removing the scattered light wavelength from the excitation light source is provided.

The particle classification apparatus according to claim 5 of the present invention is configured to irradiate a flow of an aqueous solution containing particles to be measured such as cells, chromosomes and biopolymers contained therein in a flow cell by irradiating laser light from a laser light source. In the particle classification device for detecting the scattered light and fluorescence generated by the measurement target particles and analyzing the measurement target particles based on these detected signals,
The laser light emitted from the laser light source is irradiated to the measurement target particle in the flow cell via a light beam shaping lens to excite forward scattered light and side scattered light, and at the same time, the fluorescence excitation light source. It is configured to excite fluorescence by irradiating the measurement target particle in the flow cell with more emitted light,
The forward scattered light is configured to be collected by a scattered light collecting lens and input to a forward scattered light detector to be converted into an electrical signal,
The side scattered light and fluorescent light are collected by the scattered light and fluorescent light collecting lens, reflected through the light beam splitter, and further collected by the scattered light collecting lens and input to the side scattered light detector. A fluorescent condensing lens that converts the fluorescence obtained by passing through the light beam splitter and the light having a selected wavelength by a combination of a plurality of light beam splitters and wavelength selection filters. Condensed by, input to the fluorescence detector and converted to an electrical signal,
A combination of the condenser lens and the detector that collects the forward scattered light, side scattered light, and fluorescence and converts them into electrical signals is arranged so as to block the optical path of the scattered light, and A compound lens in which a plurality of different lens elements are combined in a concentric circle or the same circle and combined with each other, and the scattered light and fluorescence imaging positions corresponding to the focal positions of the plurality of lens elements, respectively. It is characterized by comprising a combination with a plurality of detectors.

The particle classification apparatus according to claim 6 of the present invention is provided with a light emitting diode (LED) capable of emitting light in a blue region as excitation light as a fluorescence excitation light source for the measurement target particle,
Fluorescence detectors for obtaining respective required fluorescence signals through a filter for removing scattered light wavelengths with respect to scattered light and fluorescent light beams excited by irradiating the respective light sources to the measurement target particles Is provided.

  In the particle classification device according to claim 7 of the present invention, the compound lens that collects the side scattered light and the fluorescence is configured such that a plurality of lens elements each having a circle and a ring having different focal positions are combined and positioned as concentric circles. It is characterized by comprising a structure integrated in a state.

  In the particle classification device according to claim 8 of the present invention, the compound lens that condenses the forward scattered light includes a plurality of lens elements having left and right or upper and lower semicircles having different focal positions and combined positions as the same circle. It is characterized by comprising a structure integrated in a state.

In the particle classification apparatus according to claim 9 of the present invention, the compound lens that collects the forward scattered light has the irradiation with respect to the flow cell for irradiating the measurement target particles with light emitted from the light source for scattered light excitation. Consists of a configuration that is arranged at symmetrical positions on both sides of the light beam, or a configuration that is arranged on one side of the irradiation light beam with respect to the flow cell for irradiating the measurement target particles with light emitted from the light source for scattered light excitation. It is characterized by that.

  In the particle classification apparatus according to claim 10 of the present invention, the compound lens that condenses the side scattered light and the fluorescence is formed by combining a plurality of lens elements having left and right or upper and lower semicircles having different focal positions as the same circle. It is characterized by comprising a structure integrated in a positioned state.

  In the particle classification device according to an eleventh aspect of the present invention, the compound lens for condensing the side scattered light and the fluorescence is configured such that a plurality of lens elements including circles and rings having different focal positions are combined and positioned as concentric circles. It is characterized by comprising a structure integrated in a state.

In the particle classification apparatus according to claim 12 of the present invention, the compound lens that condenses the side scattered light and the fluorescence irradiates the measurement target particle with light emitted from the scattered light excitation light source or the fluorescence excitation light source. For the flow cell for irradiating the measurement target particle with the light emitted from the light source for scattering light excitation or the light source for fluorescence excitation. It is arranged on one side of the beam.

  The particle classification device according to claim 13 of the present invention is characterized in that the compound lens is provided with a pinhole for removing stray light in the center.

  The particle classification apparatus according to claim 14 of the present invention is characterized in that a collimator, a condenser lens, and a stray light removing pinhole are combined and arranged with respect to the compound lens.

  A particle classification apparatus according to a fifteenth aspect of the present invention includes an absorbance measurement unit that measures absorbance.

  According to the particle classification apparatus of the first aspect of the present invention, a light source composed of a light emitting diode (LED) capable of emitting light of a relatively short wavelength blue region as excitation light is used as a fluorescence excitation light source. By setting a new laser light source, the conventional laser light source can be made to function as a light source for exciting the scattered light for the measurement target particles, and a low-cost apparatus configuration can be obtained. The analysis of the fluorescence can be achieved easily and simply, and in particular, the analysis of the fluorescence of the particles to be measured can be achieved more appropriately and efficiently.

  According to the particle classification apparatus of the second aspect of the present invention, the laser light source is a light source for exciting scattered light with respect to the measurement target particle, and the light emitting diode capable of emitting light in the blue region as the excitation light is fluorescent with respect to the measurement target particle. As excitation light sources, signals related to forward scattered light, side scattered light, and fluorescence of the measurement target particles can be detected simultaneously and reliably, and their analysis can be achieved appropriately and efficiently.

  According to the particle classification apparatus of the third aspect of the present invention, when detecting the wavelength related to fluorescence, the wavelength related to scattered light from the scattered light excitation light source can be removed to detect a more appropriate signal related to fluorescence. it can.

  According to the particle classification device of the fourth aspect of the present invention, when detecting the wavelength related to the forward scattered light, the wavelength related to the scattered light from the fluorescence excitation light source is removed, and a more appropriate signal related to the forward scattered light is obtained. Can be detected.

  According to the particle classification apparatus of the fifth aspect of the present invention, as with the particle classification apparatus of the first aspect, the analysis of the scattered light and the fluorescence of the measurement target particle can be easily and easily achieved. In addition, the analysis of the fluorescence of the particles to be measured can be achieved more appropriately and efficiently.

  According to the particle classification device of the sixth to twelfth aspects of the present invention, a composite lens including a plurality of ring-shaped lens elements having different focal positions, and a combination of scattered light and fluorescence corresponding to the plurality of focal positions. By using a combination of a plurality of detectors respectively arranged at the image position and analyzing the measurement target particle by fluorescence detection, it is possible to easily achieve improvement in fluorescence detection sensitivity and S / N ratio. it can.

  According to the particle classification apparatus of the thirteenth and fourteenth aspects of the present invention, it is possible to effectively achieve the stray light removal by the compound lens and further improve the sensitivity and S / N ratio of fluorescence detection with respect to the measurement target particles. it can.

  According to the particle classification apparatus of claim 15 of the present invention, in addition to being able to classify and count particles, it is also possible to measure the amount of light-absorbing substance in cells (particles) by being able to measure absorbance. Can do.

  Next, embodiments of the particle classification apparatus according to the present invention will be described in detail below with reference to the accompanying drawings. For convenience of explanation, the same components as those in the particle classification apparatus shown in FIG. 3 described above will be described with the same reference numerals.

  FIG. 1 shows a schematic system configuration of an embodiment of a particle classification apparatus according to the present invention. In FIG. 1, reference numeral 10 a indicates a laser light source. The laser light source 10 a serves as a scattered light excitation light source, and emits laser light to the measurement target particle 15 in the flow cell 14 via the light beam shaping lens 12. Irradiate. In this way, the laser light applied to the measurement target particle 15 excites forward scattered light and side scattered light, respectively. The laser light irradiated to the flow cell 14 is configured such that a direct light beam incident in front of the laser light is blocked by the light beam blocker 16.

  In the particle classification apparatus of the present embodiment, a light emitting diode (LED) 30 capable of emitting light in a blue region having a relatively short wavelength as excitation light is provided as a fluorescence excitation light source for the measurement target particle. The light beam emitted from the light emitting diode 30 is applied to the measurement target particle 15 in the flow cell 14 to excite fluorescence.

  Thus, the forward scattered light excited about the measurement target particle 15 is collected by the scattered light condensing lens 18a, and then the scattered light wavelength from the fluorescence excitation light source composed of the light emitting diode 30 is removed. The light is input to the forward scattered light detector 20a via the filter 34 and converted into an electric signal. Further, the side scattered light and the fluorescence excited with respect to the measurement target particle 15 are sequentially incident on the light beam splitters 22a, 22b, 22c, and 22d that are collected by the scattered light and the fluorescence condenser lens 19a and arranged in series. Is done.

  Therefore, the side scattered light is first reflected by the light beam splitter 22a, and this reflected light beam passes through the filter 34 for removing the scattered light wavelength from the fluorescence excitation light source, and the scattered light collecting lens 18b. And is input to the side scattered light detector 20b and converted into an electric signal. In contrast, the fluorescence transmitted through the light beam splitter 22a passes through a filter 32 for removing the scattered light wavelength from the scattered light excitation light source and a filter 34 for removing the scattered light wavelength from the fluorescence excitation light source. Then, the first fluorescence set in advance is reflected by the light beam splitter 22b, and this reflected light beam is condensed by the fluorescence condenser lens 19b via the wavelength selection filter 24a and input to the fluorescence detector 26a. It is converted into an electrical signal.

  The fluorescence transmitted through the light beam splitter 22b is reflected by the second fluorescence preset by the light beam splitter 22c, and this reflected light beam is condensed by the fluorescence condenser lens 19c via the wavelength selection filter 24b. Is input to the fluorescence detector 26b and converted into an electrical signal.

  Similarly, the fluorescence that has passed through the light beam splitter 22c is reflected by the third fluorescence preset by the light beam splitter 22d, and this reflected light beam passes through the wavelength selection filter 24c and becomes a fluorescent condensing lens. The light is collected by 19d, input to the fluorescence detector 26c, and converted into an electrical signal.

  Then, the transmitted light beam composed of the fourth fluorescence transmitted through the light beam splitter 22d is condensed by the fluorescence condenser lens 19e through the wavelength selection filter 24d, and input to the fluorescence detector 26d to be converted into an electrical signal. Is done.

  With the configuration described above, according to the particle classification apparatus of the present embodiment, the scattered light excitation light source (10) and the fluorescence excitation light source (30) are individually provided for the measurement target particle 15, and excited by these light sources. Forward scattered light, side scattered light, and fluorescence can be selected and detected appropriately and reliably, and the overall configuration of the device is simplified, manufacturing costs can be reduced, and various measurement objects can be measured. Advantages that can be applied universally to particle analysis can be obtained.

  FIG. 2 shows a modification of the particle classification apparatus of the first embodiment shown in FIG. That is, in the particle classification apparatus shown in FIG. 2, when converting the forward scattered light excited about the measurement target particle 15 into an electrical signal, between the forward scattered light detector 20a and the scattered light collecting lens 18a, Between the light beam splitter 22a and the scattered light condensing lens 18b of the side scattered light detector 20b, and between the light beam splitter 22a and the filter 32 for removing the scattered light wavelength by the scattered light excitation light source, The filter 34 (see FIG. 1) for removing the scattered light wavelength from the respective fluorescence excitation light sources provided is omitted. Other configurations are the same as those of the particle classification apparatus of the embodiment shown in FIG.

  Accordingly, the same components are denoted by the same reference numerals, and detailed description thereof is omitted. Also with the particle classification device of the second embodiment configured as described above, the same function as that of the particle classification device of the first embodiment can be maintained.

  Next, FIG. 4 to FIG. 9 show the respective light condensings for condensing the forward scattered light, the side scattered light and the fluorescence, respectively, into electric signals in the particle classification apparatus shown in FIGS. A combination of lenses 18a, 18b, 19a-19c, beam splitters 22a-22c and detectors 22a, 22b, 26a, 26b are arranged so as to block the optical path of scattered light, and a plurality of lens elements having different focal positions are arranged. Compound images 40, 42, 42 ′, 44, 44 ′ that are integrated in a concentric circle or a combined position in the same circle, and imaging of the scattered light and fluorescence according to the focal positions of the plurality of lens elements It is composed of a combination of a plurality of detectors 20a, 20b, 26a to 26c arranged at respective positions, and analysis by fluorescence detection of the measurement target particles is performed, thereby detecting fluorescence. Indicating each Example in degrees and the S / N ratio improved so the particle sorting apparatus.

  FIG. 4 shows the above-described compound lens 40, 42a, 42 ′ with respect to the forward scattered light detector 20a, the side scattered light detector 20b, and the fluorescence detectors 26a-26c in the particle classification apparatus shown in FIG. The applied Example is shown. In this embodiment, the scattered light condensing lens 18a for forward scattered light is arranged so as to block the optical path of the scattered light, and a plurality of lens elements having different focal positions are integrated in a concatenated circle. The compound lens 40 (see FIGS. 10A to 10C) is used, and this compound lens 40 is forwardly moved to each focal position through a filter 34 for removing the scattered light wavelength from the fluorescence excitation light source. The scattered light detectors 20a and 20a are provided.

  In the present embodiment, the condensing lens 19a, the beam splitter 22a, and the side scattered light condensing lens 18b for the side scattered light and fluorescence are arranged so as to block the optical path of the scattered light, A compound lens 42 (see FIGS. 11 (a) to 11 (c)) in which a plurality of different lens elements are integrated together in the same circle, and a fluorescent condensing lens 19b, 19c and a beam splitter 22b, A compound lens 42 ′ with a pinhole is used as 22c, which is arranged so as to block the optical path of scattered light, and is formed by integrating a plurality of lens elements having different focal positions in a combined position in the same circle. Then, the irradiation with respect to the flow cell 14 for irradiating the measurement target particle 15 with the light emitted from the scattered light excitation light source 10a to the fluorescence excitation light source 30 through the compound lens 42 and the compound lens 42 'with a pinhole is performed. It is configured to be provided at symmetrical positions on both sides of the light beam.

  In this case, a light beam is condensed on the compound lens 42 through a filter 34 for removing the scattered light wavelength from the fluorescence excitation light source, and one of the focal positions of the compound lens 42 is focused. The side scattered light detector 20b is disposed, and the fluorescence detector 26c is disposed through the filter 32 and the wavelength selection filter 24c for removing the scattered light wavelength by the scattered light excitation light source at the other focal position. To do. Further, the compound lens 42 ′ with a pinhole is passed through a filter 34 for removing the scattered light wavelength from the fluorescence excitation light source and a filter 32 for removing the scattered light wavelength from the scattered light excitation light source. The light beam is condensed, and the fluorescence detectors 26a and 26b are disposed through the wavelength selection filters 24a and 24b, respectively, at the focal positions of the compound lens 42 'with a pinhole. It should be noted that the direct light is not blocked by the light beam blocker 16 during the actual measurement, and instead of the condenser lens or the composite lens 40, the composite lens and the collective lens are placed at an appropriate position between the flow cell and the composite lens 40. If the lens is integrated, and the light detector is directly arranged to detect the output, the absorbance of the particles flowing in the flow cell can be measured.

  According to the particle classification apparatus of the third embodiment configured as described above, it is possible to improve the sensitivity of fluorescence detection and the S / N ratio when analyzing the measurement target particles by fluorescence detection. In this embodiment, instead of the composite lens 42 (see FIGS. 11A to 11C) and its composite lens 42 'with a pinhole, a composite lens 44 (FIGS. 12A to 12C) is used. Reference] or a compound lens 44 ′ with a pinhole (see FIGS. 13A to 13C) can be applied.

  FIG. 5 shows the above-described compound lenses 40, 42, and 42 ′ with respect to the forward scattered light detector 20a, the side scattered light detector 20b, and the fluorescence detectors 26a to 26c in the particle classification apparatus shown in FIG. Another embodiment applied is shown. That is, in the present embodiment, the scattered light condensing lens 18a with respect to the forward scattered light is arranged so as to block the optical path of the scattered light, and a plurality of lens elements having different focal positions are combined and concentrically positioned. An integrated composite lens 40 (see FIGS. 10A to 10C) is used, and forward scattered light detectors 20a and 20a are provided at respective focal positions of the composite lens 40.

  Further, in the present embodiment, as in the third embodiment, as the condensing lens 19a, the beam splitter 22a, and the side scattered light condensing lens 18b for side scattered light and fluorescence, the optical path of the scattered light is blocked. A compound lens 42 (see FIGS. 11 (a) to 11 (c)) in which a plurality of lens elements arranged in different positions and integrated as a single circle are combined together and a fluorescent condensing lens 19b. 19c and beam splitters 22b and 22c are arranged so as to block the optical path of the scattered light, and a plurality of lens elements having different focal positions are integrated in a state where they are combined as the same circle and integrated with a pinhole A lens 42 'is used. Then, the irradiation with respect to the flow cell 14 for irradiating the measurement target particle 15 with the light emitted from the scattered light excitation light source 10a to the fluorescence excitation light source 30 through the compound lens 42 and the compound lens 42 'with a pinhole is performed. It is configured to be provided at symmetrical positions on both sides of the light beam.

  In this case, the side scattered light detector 20b is arranged for one focal position of the compound lens 42, and the scattered light wavelength by the scattered light excitation light source is removed for the other focal position. The fluorescence detector 26c is arranged through the filter 32 and the wavelength selection filter 24c. Further, a light beam is condensed on the compound lens 42 'with a pinhole through a filter 32 for removing the scattered light wavelength by the scattered light excitation light source, and each of the compound lenses 42' with the pinhole is collected. Fluorescence detectors 26a and 26b are arranged with respect to the focal position via wavelength selection filters 24a and 24b, respectively. It should be noted that the direct light is not blocked by the light beam blocker 16 during the actual measurement, and instead of the condenser lens or the composite lens 40, the composite lens and the collective lens are placed at an appropriate position between the flow cell and the composite lens 40. If the lens is integrated, and the light detector is directly arranged to detect the output, the absorbance of the particles flowing in the flow cell can be measured.

  According to the particle sorting apparatus of the fourth embodiment configured as described above, it is possible to improve the sensitivity of fluorescence detection and the S / N ratio when analyzing the measurement target particles by fluorescence detection. In this embodiment, instead of the compound lens 42 (see FIGS. 11 (a) to 11 (c)) and the compound lens 42 'with a pinhole, a compound lens 44 (see FIGS. 12 (a) to (c)). Or a compound lens 44 ′ with a pinhole (see FIGS. 13A to 13C) can be applied.

  6 and 7 (a) and 7 (b) show the forward scattered light detector 20a, the side scattered light detector 20b, and the fluorescence detectors 26a to 26c in the particle classification apparatus shown in FIG. An embodiment to which the compound lenses 40 and 44 'are applied is shown. That is, in the present embodiment, the scattered light condensing lens 18a with respect to the forward scattered light is arranged so as to block the optical path of the scattered light, and a plurality of lens elements having different focal positions are combined and concentrically positioned. An integrated composite lens 40 (see FIGS. 10A to 10C) is used, and forward scattered light detectors 20a and 20a are provided at respective focal positions of the composite lens 40.

  In the present embodiment, the condensing lens 19a, the beam splitter 22a, and the side scattered light condensing lens 18b for the side scattered light and fluorescence are arranged so as to block the optical path of the scattered light, Using a compound lens 44 ′ with a pinhole (see FIGS. 13A to 13C) in which a plurality of different lens elements are integrated in a state where they are coupled as the same circle, the compound lens 44 with a pinhole is used. ′ Is arranged on one side of the irradiation light beam with respect to the flow cell 14 for irradiating the measurement target particle 15 with light emitted from the scattered light excitation light source 10a to the fluorescence excitation light source 30 (FIG. 6).

  In this case, the side scattered light detector 20b is arranged for one focal position of the compound lens 44 'with a pinhole, and the scattered light wavelength by the light source for scattered light excitation is arranged for the other focal position. The fluorescence detector 26c is arranged through a filter 32 for removing [see FIG. 7 (a)]. It should be noted that the direct light is not blocked by the light beam blocker 16 during the actual measurement, and instead of the condenser lens or the composite lens 40, the composite lens and the collective lens are placed at an appropriate position between the flow cell and the composite lens 40. If the lens is integrated, and the light detector is directly arranged to detect the output, the absorbance of the particles flowing in the flow cell can be measured.

  According to the particle sorting apparatus of the fifth embodiment configured as described above, as shown in FIG. 7B, the fluorescence excitation light source 30, the side scattered light detector 20b, and the fluorescence are formed on one substrate 46. The detector 26c can be disposed, and the entire apparatus can be easily reduced in size and cost. Further, when analyzing the measurement target particle by fluorescence detection, it is possible to improve the sensitivity and S / N ratio of fluorescence detection.

  FIG. 8 shows an embodiment in which the compound lenses 40 and 42 are applied to the forward scattered light detector 20a, the side scattered light detector 20b, and the fluorescence detectors 26a to 26c in the particle classification apparatus shown in FIG. An example is given. That is, in the present embodiment, the scattered light condensing lens 18a with respect to the forward scattered light is arranged so as to block the optical path of the scattered light, and a plurality of lens elements having different focal positions are combined and concentrically positioned. An integrated composite lens 40 (see FIGS. 10A to 10C) is used, and forward scattered light detectors 20a and 20a are provided at respective focal positions of the composite lens 40.

  In the present embodiment, the condensing lens 19a, the beam splitter 22a, and the side scattered light condensing lens 18b for the side scattered light and fluorescence are arranged so as to block the optical path of the scattered light, A first compound lens 42 (see FIGS. 11 (a) to 11 (c)) formed by integrating a plurality of different lens elements in a combined state in the same circle, the fluorescent condensing lenses 19b and 19c, and beam splitting As the devices 22b and 22c, a second compound lens 42 is used which is arranged so as to block the optical path of the scattered light, and is formed by integrating a plurality of lens elements having different focal positions in a combined position in the same circle. . The first compound lens 42 and the second compound lens 42 are applied to the flow cell 14 for irradiating the measurement target particle 15 with light emitted from the scattered light excitation light source 10a to the fluorescence excitation light source 30. It is configured to be provided at symmetrical positions on both sides of the irradiation light beam.

  In this case, the side scattered light detector 20b is arranged for one focal position of the first compound lens 42, and the scattered light wavelength by the scattered light excitation light source is set for the other focal position. The fluorescence detector 26c is arranged through the filter 32 and the wavelength selection filter 24c for removal. Further, a light beam is condensed on the second compound lens 42 through a filter 32 for removing the scattered light wavelength by the scattered light excitation light source, and each focal position of the compound lens 42 is collected. , Fluorescence detectors 26a and 26b are arranged through wavelength selection filters 24a and 24b, respectively. It should be noted that the direct light is not blocked by the light beam blocker 16 during the actual measurement, and instead of the condenser lens or the composite lens 40, the composite lens and the collective lens are placed at an appropriate position between the flow cell and the composite lens 40. If the lens is integrated, and the light detector is directly arranged to detect the output, the absorbance of the particles flowing in the flow cell can be measured.

  According to the particle classification apparatus of the sixth embodiment configured as described above, it is possible to improve the sensitivity of fluorescence detection and the S / N ratio when analyzing the measurement target particles by fluorescence detection. In this embodiment, instead of the compound lens 42 (see FIGS. 11 (a) to 11 (c)), the compound lens 42 'with a pinhole or the compound lens 44 (see FIGS. 12 (a) to (c)). Or a compound lens 44 ′ with a pinhole (see FIGS. 13A to 13C) can be applied.

  FIG. 9 shows an embodiment in which the compound lenses 40 and 42 are applied to the forward scattered light detector 20a, the side scattered light detector 20b, and the fluorescence detectors 26a to 26c in the particle classification apparatus shown in FIG. An example is given. That is, in the present embodiment, the scattered light condensing lens 18a with respect to the forward scattered light is arranged so as to block the optical path of the scattered light, and a plurality of lens elements having different focal positions are combined and concentrically positioned. An integrated composite lens 40 (see FIGS. 10A to 10C) is used, and forward scattered light detectors 20a and 20a are provided at respective focal positions of the composite lens 40.

  In the present embodiment, the condensing lens 19a, the beam splitter 22a, and the side scattered light condensing lens 18b for the side scattered light and fluorescence are arranged so as to block the optical path of the scattered light, A first compound lens 42 (see FIGS. 11 (a) to 11 (c)) formed by integrating a plurality of different lens elements in a combined state in the same circle, the fluorescent condensing lenses 19b and 19c, and beam splitting As the devices 22b and 22c, a second compound lens 42 is used which is arranged so as to block the optical path of the scattered light, and is formed by integrating a plurality of lens elements having different focal positions in a combined position in the same circle. . The first compound lens 42 and the second compound lens 42 are applied to the flow cell 14 for irradiating the measurement target particle 15 with light emitted from the scattered light excitation light source 10a to the fluorescence excitation light source 30. It is configured to be provided at symmetrical positions on both sides of the irradiation light beam.

  In this case, the side scattered light detector 20b is arranged for one focal position of the first compound lens 42, and the scattered light wavelength by the scattered light excitation light source is set for the other focal position. The fluorescence detector 26c is arranged through the filter 32 and the wavelength selection filter 24c for removal. For the second compound lens 42, a combination of a collimator 50a, a condensing lens 52, a pinhole plate 54, and a collimator 50b, and a filter 32 for removing the scattered light wavelength from the scattered light excitation light source The light beam is condensed via the wavelength selection filters 24 a and 24 b, and the fluorescence detectors 26 a and 26 b are arranged for the respective focal positions of the compound lens 42. It should be noted that the direct light is not blocked by the light beam blocker 16 during the actual measurement, and instead of the condenser lens or the composite lens 40, the composite lens and the collective lens are placed at an appropriate position between the flow cell and the composite lens 40. If the lens is integrated, and the light detector is directly arranged to detect the output, the absorbance of the particles flowing in the flow cell can be measured.

  According to the particle classification apparatus of the seventh embodiment configured as described above, the sensitivity of fluorescence detection and the S / N ratio can be improved in the analysis by the fluorescence detection of the measurement target particles. In this embodiment, instead of the compound lens 42 (see FIGS. 11A to 11C), a compound lens 42 'with a pinhole or a compound lens 44 (FIGS. 12A to 12C) is used. Reference] and a compound lens 44 ′ with a pinhole (see FIGS. 13A to 13C) can be applied.

  Next, characteristics and manufacturing methods of the compound lenses 40, 42, 44, and 44 'used in the particle classification apparatuses of the third to seventh embodiments will be described.

  FIGS. 10A to 10C are compound lenses which are arranged so as to block the optical path of scattered light, and are integrated in a state where a plurality of lens elements having different focal positions are combined and positioned as concentric circles or the same circle. 40 characteristics and a manufacturing method thereof are shown.

  That is, the compound lens 40 shown in FIGS. 10A to 10C is cut or cut into a circular shape from a mold A that can mold the Fresnel lens A, and a mold A ′ (mold A ′) as a first lens element. [See FIG. 10 (a)]. Further, a mold B ′ (mold B ′) as a ring-shaped second lens element provided with a hole capable of incorporating the mold A ′ is cut or cut out from the mold B (mold B) capable of molding the mold B ′. [See FIG. 10 (b)]. By incorporating the mold A ′ as the first lens element thus obtained into the mold B ′ as the second lens element, the compound lens 40 having the mold surface as a concentric circle can be molded. Yes (see FIG. 10C). In this case, the mold A (moldA) and the mold B (moldB) may be the same, or may be molds for molding Fresnel lenses having different focal lengths or different principal axes. In addition, the compound lens 40 thus molded can be set so as to obtain a focal position corresponding to each lens element as shown in the figure.

  11A to 11C show a composite lens 42 that is arranged so as to block the optical path of scattered light, and is formed by integrating a plurality of lens elements having different focal positions in a combined position in the same circle. The characteristics and the manufacturing method are shown.

  That is, the compound lens 42 shown in FIGS. 11A to 11C is molded from a mold A (mold A) capable of molding the Fresnel lens A into a left semicircular shape or cut into a left semicircular shape, and the mold A as the first lens element. ′ (Mold A ′) is molded (see FIG. 11A). Further, a mold B ′ as a second lens element is molded by cutting or cutting from the mold B to a right semicircular shape from a circular shape (see FIG. 11B). By combining the mold A ′ as the first lens element and the mold B ′ as the second lens element obtained in this manner, the compound lens 42 composed of the mold surface as the same circle is obtained. It can be molded (see FIG. 11C). In this case, the mold A (moldA) and the mold B (moldB) may be the same, or may be molds for molding Fresnel lenses having different focal lengths or different principal axes. Further, the compound lens 42 thus molded can be set so as to obtain a focal position corresponding to each lens element as shown in the figure.

  FIGS. 12A to 12C show another compound lens that is arranged so as to block the optical path of scattered light, and is formed by integrating a plurality of lens elements having different focal positions in a combined position in the same circle. The characteristics of 44 and the manufacturing method thereof are shown.

  That is, the compound lens 44 shown in FIGS. 12A to 12C is formed by cutting or cutting a mold A (mold A) capable of molding the Fresnel lens A into a semicircular shape that is higher than a circle and cutting it as a first lens element. ′ (Mold A ′) is molded (see FIG. 12A). Further, a mold B ′ (mold B ′) as a second lens element is formed by cutting or cutting from the mold B (mold B) into a lower semicircular shape rather than a circular shape (see FIG. 12B). By combining the mold A ′ as the first lens element and the mold B ′ as the second lens element obtained in this manner, the compound lens 44 composed of the mold surface as the same circle is obtained. It can be molded (see FIG. 12C). In this case, the mold A (moldA) and the mold B (moldB) may be the same, or may be molds for molding Fresnel lenses having different focal lengths or different principal axes. Further, the compound lens 44 molded in this way can be set so as to obtain a focal position corresponding to each lens element as shown in the figure.

  FIGS. 13A to 13C show characteristics of a compound lens 44 ′ provided with a pinhole p in the compound lens 44 shown in FIGS. 12A to 12C and a manufacturing method thereof.

That is, the compound lens 44 ′ shown in FIGS. 13A to 13C is basically the same as the method of manufacturing the compound lens 44 shown in FIGS. 12A to 12C, and the first lens element is used. When the mold A ′ is coupled to the mold B ′ as the second lens element, a pinhole p is provided at the coupling center.
Accordingly, although not shown in the drawings, the above-described compound lens 42 shown in FIGS. 11A to 11C also uses the mold A ′ as the first lens element as the mold as the second lens element, as described above. When coupled to B ′, a compound lens 42 ′ having a pinhole p at the coupling center can be molded.

  The preferred embodiments of the present invention have been described above, but the present invention is not limited to the above-described embodiments, and many design changes can be made without departing from the spirit of the present invention. .

1 is a schematic system configuration diagram showing a first embodiment of a particle classification device according to the present invention. It is a schematic system block diagram which shows 2nd Example of the particle classification apparatus which concerns on this invention. It is a schematic system block diagram which shows the structural example of the conventional particle classification apparatus. It is a schematic system block diagram which shows the 3rd Example of the particle classification device based on this invention which applied the combination of the compound lens and the some detector to the particle classification device shown in FIG. It is a schematic system block diagram which shows the 4th Example of the particle classification device based on this invention which applied the combination of the compound lens and the some detector to the particle classification device shown in FIG. It is a schematic system block diagram which shows 5th Example of the particle classification device based on this invention which applied the combination of the compound lens and the some detector to the particle classification device shown in FIG. (A) And (b) is a schematic system block diagram which shows the example of each structure arrangement | positioning of the fluorescence detector in 5th Example of the particle classification device based on this invention shown in FIG. It is a schematic system block diagram which shows the 6th Example of the particle classification device based on this invention which applied the combination of the compound lens and the some detector to the particle classification device shown in FIG. It is a schematic system block diagram which shows the 7th Example of the particle classification device based on this invention which applied the combination of the compound lens and the some detector to the particle classification device shown in FIG. (A), (b) and (c) is explanatory drawing which shows the characteristic of the compound lens [40] applied to the particle classification apparatus based on this invention, and its manufacturing process, respectively. (A), (b) and (c) is explanatory drawing which shows the characteristic of another compound lens [42a-c] applied to the particle classification device based on this invention, and its manufacturing process, respectively. (A), (b) and (c) is explanatory drawing which shows the characteristic of another compound lens [44a-c] applied to the particle classification device based on this invention, and its manufacturing process, respectively. (A), (b), and (c) are explanatory views showing the characteristics of still another compound lens [44a ′ to c ′] applied to the particle classification apparatus of the present invention and the manufacturing process thereof.

Explanation of symbols

10a Laser light source of the present invention (for scattered light excitation)
10b Conventional laser light source (for scattered light detection and fluorescence excitation)
12 Light beam shaping lens 14 Flow cell 15 Particle 16 to be measured Light beam blocker 18a, 18b Scattered light condensing lens 19a Scattered light and fluorescent condensing lenses 19b-19e Fluorescent condensing lens 20a Forward scattered light detector 20b Side scattered light Detectors 22a to 22d Light beam splitters 24a to 24d Wavelength selection filters 26a to 26d Fluorescence detector 30 Light emitting diode (fluorescence excitation light source)
32 filter (scattered light wavelength removal by light source for excitation of scattered light)
33 filter (scattered light wavelength removal by scattered light excitation light source or scattered light wavelength removal from fluorescence excitation light source)
34 Filter (Removes scattered light wavelength from light source for fluorescence excitation)
40 Compound Lens (Concentric Fresnel Lens)
42 Compound lens (horizontal division type Fresnel lens)
Compound lens with 42 'pinhole (horizontal division type Fresnel lens)
44 Compound lens (Vertical division type Fresnel lens)
Compound lens with 44 'pinhole (Vertical division type Fresnel lens)
46 Substrate 50a, 50b Collimator 52 Condensing lens 54 Pinhole plate p Pinhole

Claims (15)

The scattered light and fluorescence generated by the measurement target particles are detected by irradiating the flow of an aqueous solution containing the measurement target particles such as cells, chromosomes, and biopolymers contained in the flow cell with a laser light source. Then, in the particle classification device for analyzing the measurement target particles based on these detected signals,
As a fluorescence excitation light source for the measurement target particles, a light emitting diode (LED) capable of emitting blue region light as excitation light is provided,
Fluorescence detectors for obtaining respective required fluorescence signals through a filter for removing scattered light wavelengths with respect to scattered light and fluorescent light beams excited by irradiating the respective light sources to the measurement target particles A particle classification apparatus characterized by comprising: The laser light emitted from the laser light source is irradiated to the measurement target particle in the flow cell via a light beam shaping lens to excite forward scattered light and side scattered light, and at the same time, the fluorescence excitation light source. It is configured to excite fluorescence by irradiating the measurement target particle in the flow cell with more emitted light,
The forward scattered light is configured to be collected by a scattered light collecting lens and input to a forward scattered light detector to be converted into an electrical signal,
The side scattered light and fluorescent light are collected by the scattered light and fluorescent light collecting lens, reflected through the light beam splitter, and further collected by the scattered light collecting lens and input to the side scattered light detector. A fluorescent condensing lens that converts the fluorescence obtained by passing through the light beam splitter and the light having a selected wavelength by a combination of a plurality of light beam splitters and wavelength selection filters. The particle classification apparatus according to claim 1, wherein the particle classification apparatus is configured so as to collect the light and to input to a fluorescence detector and convert it into an electrical signal.   Scattering by a light source for exciting scattered light is provided at the forefront of the combination of the plurality of light beam splitters and wavelength selection filters with respect to a fluorescence detector that converts the fluorescence obtained through the light beam splitter into an electrical signal. The particle classification apparatus according to claim 2, further comprising a filter for removing light wavelengths.   A filter for removing the wavelength of the scattered light from the fluorescence excitation light source between the scattered light condensing lens and the forward scattered light detector with respect to the forward scattered light detector that converts the forward scattered light into an electrical signal. The particle classification apparatus according to claim 3, wherein: The scattered light and fluorescence generated by the measurement target particles are detected by irradiating the flow of an aqueous solution containing the measurement target particles such as cells, chromosomes, and biopolymers contained in the flow cell with a laser light source. Then, in the particle classification device for analyzing the measurement target particles based on these detected signals,
The laser light emitted from the laser light source is irradiated to the measurement target particle in the flow cell via a light beam shaping lens to excite forward scattered light and side scattered light, and at the same time, the fluorescence excitation light source. It is configured to excite fluorescence by irradiating the measurement target particle in the flow cell with more emitted light,
The forward scattered light is collected by a scattered light condensing lens and is input to a forward scattered light detector to be converted into an electrical signal,
The side scattered light and fluorescent light are collected by the scattered light and fluorescent light collecting lens, reflected through the light beam splitter, and further collected by the scattered light collecting lens and input to the side scattered light detector. A fluorescent condensing lens that converts the fluorescence obtained by passing through the light beam splitter and the light having a selected wavelength by a combination of a plurality of light beam splitters and wavelength selection filters. Condensed by, input to the fluorescence detector and converted to an electrical signal,
A combination of the condenser lens and the detector that collects the forward scattered light, side scattered light, and fluorescence and converts them into electrical signals is arranged so as to block the optical path of the scattered light, and A compound lens in which a plurality of different lens elements are combined in a concentric circle or the same circle and combined with each other, and the scattered light and fluorescence imaging positions corresponding to the focal positions of the plurality of lens elements, respectively. A particle classification device comprising a combination with a plurality of detectors. As a fluorescence excitation light source for the measurement target particles, a light emitting diode (LED) capable of emitting blue region light as excitation light is provided,
Fluorescence detectors for obtaining respective required fluorescence signals through a filter for removing scattered light wavelengths with respect to scattered light and fluorescent light beams excited by irradiating the respective light sources to the measurement target particles The particle classification apparatus according to claim 5, further comprising:   6. The compound lens for condensing the forward scattered light has a configuration in which a plurality of lens elements composed of a circle and a ring having different focal positions are integrated in a concatenated position. Or the particle classification device of 6.   The compound lens for condensing the forward scattered light has a configuration in which a plurality of lens elements composed of right and left or upper and lower semicircles having different focal positions are integrated in a combined position in the same circle. Item 7. A particle classification apparatus according to Item 5 or 6. The compound lens that condenses the forward scattered light is arranged at symmetrical positions on both sides of the irradiation light beam with respect to the flow cell for irradiating the light to be measured with light emitted from the light source for scattered light excitation. The particle classification according to claim 7 or 8, comprising a configuration in which a flow cell for irradiating measurement target particles with light emitted from a scattered light excitation light source is arranged on one side of the irradiation light beam. apparatus.   The compound lens that condenses the side scattered light and the fluorescence has a structure in which a plurality of lens elements having left and right or upper and lower semicircles having different focal positions are integrated in a combined position in the same circle. The particle classification apparatus according to claim 5 or 6.   The compound lens that condenses the side scattered light and the fluorescence has a structure in which a plurality of lens elements composed of a circle and a ring having different focal positions are integrated in a concatenated circle. The particle classification apparatus according to claim 5 or 6. The compound lens for condensing the side scattered light and the fluorescence is arranged on both sides of the irradiation light beam with respect to the flow cell for irradiating the measurement target particle with the light emitted from the scattered light excitation light source or the fluorescence excitation light source. Each of the irradiation light beams is arranged at one side of the irradiation light beam with respect to the flow cell for irradiating the measurement target particle with light emitted from each of the symmetrical positions or the light emitted from the scattered light excitation light source or the fluorescence excitation light source. Item 10. The particle classification device according to Item 10 or 11.   13. The particle classification apparatus according to claim 7, wherein the compound lens is provided with a stray light removing pinhole at a central portion.   The particle classification apparatus according to claim 9, wherein a collimator, a condenser lens, and a stray light removing pinhole are combined and arranged with respect to the compound lens.   The particle classification apparatus according to claim 5 or 6, further comprising an absorbance measurement unit that measures absorbance based on the forward scattered light.