JPH1123447A - Particle-analyzing device, compound lens integrating lens-element with a plurality of focal point - Google Patents

Abstract

PROBLEM TO BE SOLVED: To enable a particle analysis with high measurement accuracy by connecting and integrating a plurality of lens elements with different focal points 1 to form a compound lens and separating and detecting light with different scattering angles. SOLUTION: The first detector 16 is arranged at the image-forming location of light which has passed through a convex lens element 15a with the shortest focal length, the second detector 17 is arranged at the image forming location of light which has passed through the first lens element 15b with a medium focal length, and the third detector 18 is arranged at the image forming location of light which has passed through the second lens element 15c with the longest focal length. Then the outputs of the detectors 16 to 18 are given to an analyzing device 9 to analyze particles. Further, the first detector 16 is provided with a pinhole so as to match the center of a light receiving plane in order not to detect light displaced from the center. The analyzed results based on the outputs of the second and third detectors 17 and 18 are outputted to an output device 19. In addition, analytical and classifying processes performed on the basis of signals from different scattering angles are performed as before.

Description

DETAILED DESCRIPTION OF THE INVENTION

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a particle analyzing apparatus represented by a flow cytometer and a compound lens in which a lens element having a plurality of focal positions is integrated.
More specifically, the present invention relates to an apparatus that can classify particles (samples) such as cells and blood cells based on scattering angle information of forward scattered light.

[0002]

2. Description of the Related Art Flow cytometers include a cell sorter, a cell analyzer, and a particle analyzer (blood analyzer) incorporating them, and the like. Light is emitted to particles flowing in the flow cell and particles falling from the flow cell. Irradiation, forward scattered light and side scattered light generated at that time are detected, and particles such as cells and blood cells are classified based on the detected light information.

It is known that among forward scattered lights, the scattering angle varies depending on the type of particles. Therefore, there has been proposed an apparatus for classifying particles by dividing forward scattered light into a plurality of regions having different scattering angles and concentrating and measuring the divided regions. One example is disclosed in
71509, which is an apparatus for optically detecting each cell contained in leukocytes and accurately classifying cells having a low content. FIGS. 15 and 1 show a structure for detecting scattered light in the device disclosed in this publication.
6, as shown in FIG.

As shown in the cross-sectional views of the flow cell used in the flow cytometer shown in FIGS. 16 and 17, the inside of the flow cell 2 is a sample in which a sample (particles) flows into a central portion of a sheath liquid 2a flowing in a sheath shape. A tube 2c is located, and a sample flow 2b containing particles is sequentially supplied to the thin tube portion so as to be wrapped in a sheath liquid at the distal end. Since the laser beam a is irradiated on the narrow tube portion, the particles 7 passing therethrough become scattering sources, and forward scattered light b, c, d, side scattered light e, f, g, (Not shown) and backscattered light h, j.

As shown in FIG. 15, a laser beam emitted from a light source 1 passes through a flow cell 2 and is blocked by an immovable obsculator 11. The forward scattered light beams b, c, d, etc. are converted into parallel light beams by a collimator lens 3 installed in front of the flow cell 2 and are irradiated on a front mirror 4. The mirror 4 has a hole 4 in the center.
The light having a small scattering angle among the forward scattered light is formed in the hole 4a, and is collected by the first condenser lens 5 on the first detector 6. Further, of the forward scattered light, the light having a large scattering angle is radiated to the periphery of the mirror 4 and is reflected there to change the optical path by 90 degrees, and is condensed on the second detector 6 'by the second condenser lens 5'. Is done. Each of the first and second detectors 6 and 6 ′ is configured by a photoelectric conversion element such as a photodiode, converts the received light into an electric signal corresponding to the intensity of the received light, and sends the electric signal to the analyzer 9.

That is, the first detector 6 detects forward small-angle scattered light intensity data having a small scattering angle, and the second detector 6 '.
Detects forward large-angle scattered light intensity data having a large scattering angle. Then, the analyzer 9 calculates and stores forward small-angle scattered light intensity data and forward large-angle scattered light intensity data from each input detection signal, and calculates two-dimensional coordinate data of forward small-angle and forward large-angle scattered light intensity. I do. The analyzing device 9 sequentially outputs the calculated forward small-angle scattered light intensity data and forward-large-angle scattered light intensity data to an external display device (not shown), and displays the light intensity scatter distribution of two-dimensional coordinates on a screen. It is displayed as a figure (scattergram).

[0007] The following directions are known for the scattered light scattered at the time of laser beam irradiation. That is, when the size of the particles is large with respect to the wavelength of the laser light, it becomes forward scattered light, and when the size of the particles is approximately the same as the wavelength of the laser light, it becomes forward and side scattered light, and the size of the particles becomes large. It is known that when the wavelength is small with respect to the wavelength of the laser light, the light becomes forward, side, and back scattered light.

Accordingly, in order to classify the particles more accurately in an actual apparatus, as shown in FIGS. 16 and 17, for the forward scattered light b, c, and d, the collimator lens 3, the mirror having the hole, and the like. 4, an optical system 8 having a condenser lens 5
And a detector 6 for detecting each light intensity. Similarly, the optical system 8 and the detector 6 are provided for the side scattered lights e, f, and g, and the optical system 8 and the detector 6 are provided for the back scattered light h and i.

FIG. 18 shows another apparatus. The illustrated device does not separate the light into a plurality of regions having different scattering angles by the above-described mirror 4 having a hole, but separates the light using an optical fiber 10. That is, the forward scattered light emitted from the particles in the flow cell 2 is applied to the light receiving surface of the optical fiber 10. This light receiving surface is, as shown in FIG. 2B, an optical fiber group 10 for receiving small forward angled scattered light at the center.
a and the surrounding optical fiber group 1 for receiving large angle forward scattered light
0b is arranged. Then, as shown in FIG. 3A, the emission side of the optical fiber group 10a is connected to the first detector 6, and the emission side of the optical fiber group 10b is connected to the second detector 8. Thereby, the scattered light applied to the light receiving surface of the optical fiber 10 is supplied to one of the detectors 6 and 8 by a predetermined optical fiber group according to the scattering angle. The outputs of the first and second detectors 6 and 8 are sent to an analyzer in the same manner as in FIG. 6, and predetermined signal processing is performed.

[0010]

However, the above-described conventional apparatus has the following problems. That is, in the apparatus shown in FIG. 15, a collimator lens 3, a mirror 4 with a hole, and a condenser lens 5 are required as an optical system for allowing each of the detectors 6 and 6 'to receive light. Adjustment is complicated and complicated. Further, the occupied area increases, which hinders miniaturization of the entire apparatus. Further, as shown in FIG. 16 and FIG. 17, the area to be divided is increased, and furthermore, in the device for detecting the side scattered light and the back scattered light, the number of mirrors having holes equal to "the number of scattered light to be detected-1" The same number of condensing lenses as the number of scattered lights is required, which causes an increase in the number of components, and the above-mentioned problem becomes more remarkable.

On the other hand, the apparatus shown in FIG. 18 does not require a collimator lens or a condenser lens, and requires only optical fibers as the number of components. Moreover, even if the area to be divided is increased, only the number of fiber bundles is increased.
6, there is little increase in the number of parts and the occupied area as shown in FIG. However, since the light receiving surface of each optical fiber bundle must be arranged concentrically, and the light exits on the opposite side must be grouped in optical fiber bundle units, the routing of the optical fiber is complicated, and the number of divisions is limited. Also,
In the first place, the loss is large in the case of an optical fiber. In other words, not only the loss while passing through the optical fiber, but also the light applied to the clad portion is not transmitted, and eventually the light applied to the light receiving surface of the optical fiber 10 shown in FIG. The light reaching the vessel is less than 50%, and the light use efficiency is poor.

SUMMARY OF THE INVENTION The present invention has been made in view of the above background, and has as its object to solve the above-mentioned problems and to separate light having different scattering angles with a simple configuration. Can be easily adjusted, the light utilization rate is high,
Moreover, even if the number of regions to be divided increases, the number of components of the optical system for separating light also does not increase, and the occupied area does not increase, so that the space due to the increase in the number of detectors can be reduced. A compound lens which can easily and accurately align the position of a compound lens which is a main part of the invention and has a high measurement accuracy and a lens element having a plurality of focal positions suitable for use therein. It is to provide a lens.

[0013]

In order to achieve the above-mentioned object, a particle analyzer according to the present invention separates light having different scattering angles from scattered light generated by irradiating particles with light. In the particle analyzer that is configured to perform the classification of the particles by performing detection, a plurality of ring-shaped lens elements having different focal positions, which are arranged so as to block the optical path of the scattered light, are positioned on concentric circles. And a detector for detecting each scattering angle (in the embodiment, the first lens in the first embodiment) disposed at a position where the scattered light is imaged in accordance with the focal positions of the plurality of lens elements. To third detectors 36 to 38) and analysis means for classifying the particles based on detection outputs of the plurality of detectors (claim 1). The present invention relates to the fourth and fifth aspects.
This is realized by the embodiment described above.

Further, in the particle analyzer which classifies the particles by separating and detecting those having different scattering angles among the scattered lights generated by irradiating the particles with the light, A plurality of ring-shaped lens elements having different focal positions, which are arranged so as to block an optical path, are positioned concentrically, and a compound lens integrated with a positioning element provided in a central portion; Detectors (in the embodiment, second and third detectors 17 and 18 in the embodiment) each for detecting a scattering angle, which are respectively arranged at the image forming positions of the scattered light according to the focal positions of the lens elements of the above, and the positioning element A position detector (in the embodiment, the first detector 16) capable of detecting light that has passed through, and analysis means for classifying the particles based on detection outputs of the plurality of detectors. The positioning element is located at a predetermined position of the positioning detector installed forward when at least the center of the compound lens coincides with the optical axis and the compound lens is orthogonal to the optical axis. Alternatively, a configuration may be adopted in which a predetermined amount of light can be received (claim 2). The present invention is realized by the first to third embodiments.

Note that the image forming position corresponding to the focal position where each detector is installed is not necessarily the focal position. That is, since the scattered light applied to the composite lens is not a parallel light flux, no image is formed at the focal position. However, the position where the scattered light is generated is a particle flowing in the flow cell or a particle falling from the flow cell, and in any case, since the relative positional relationship with the complex lens is known,
The imaging position is uniquely determined.

Thus, the scattered light is applied to a predetermined position of the compound lens. That is, forward small-angle scattered light having a small scattering angle is applied to a ring-shaped lens element located at the center side of the compound lens, and large forward-angle scattered light having a large scattering angle is applied to lens elements located around the compound lens. Is done. Since each lens element has a different focal position, the light applied to each lens element is imaged at a different position and detected by a detector.

Further, in the invention according to claim 2, since the positioning element is provided at the center of the compound lens, whether the posture of the compound lens has reached a desired state is determined by monitoring the output of the positioning detector. , Can be easily determined. Therefore, positioning and attitude control can be easily performed.

Note that the predetermined light amount is a light amount that is equal to or higher than a predetermined reference, and the predetermined reference is a predetermined value whose reference value itself is larger than 0, such as one using a pinhole as a positioning element. In some cases, as in the case where a photodiode with a pinhole is used as a detector and a convex lens is used as a positioning element, the reference value is 0 and whether or not the reference value is larger than 0 (whether there is light reception or not). ) Is also included.

If the components of each of the above-mentioned particle analyzers are more specifically limited, the following can be achieved. That is, first, as the positioning element, various elements can be used as long as they satisfy the condition of claim 2, and for example, a convex lens or a pinhole can be used.

The complex lens may be an ordinary lens, but it is preferable to use a Fresnel lens because it can be easily designed and manufactured and can be easily handled.

As a structure for making the focal positions different, for example, a plurality of lens elements having different focal lengths are joined, a plurality of lens elements having different main axes are joined, or a combination thereof. In addition, various structures can be adopted.

Furthermore, the boundary between the plurality of lens elements and / or the boundary between the positioning element and the lens element may be covered with an annular mask material. When covered with a mask material in this way, light is condensed based on the light of the lens element portion with a stable composition, so it is difficult for noise components to ride on the signal detected by the detector, enabling more accurate measurement. Become.

Further, a light-shielding member for shielding direct light emitted from a light source is provided at a predetermined position between the flow cell to which the particles are supplied and the compound lens, and the light-shielding member is provided with the positioning element. When the composite lens is adjusted by using it, it may be retractable. That is, at the time of measurement, if direct light and forward scattered light from a light source having a large light amount are received by the detector together, information on the forward scattered light is canceled out. Therefore, in a normal flow cytometer, a light-blocking member is provided at a predetermined position so that direct light does not irradiate the complex lens. On the other hand, when the attitude of the compound lens is being adjusted, light must be applied to the central positioning element of the compound lens. However, if the light blocking member is provided, the positioning element cannot be irradiated with light. Therefore, it is preferable that the light-shielding member can be retracted and that the light-shielding member can be retracted during the adjustment of the complex lens so that the positioning element can be irradiated with light so that both positioning and measurement can be performed reliably.

In this evacuation, for example, a light-blocking member may be attached to a slide rail or the like and moved along the slide rail. In this case, it can be automatically connected to driving means such as a motor.
Moreover, you may make it remove manually from an installation position. As an example of this manual removal, for example, the light shielding member can be removably attached to a holder for mounting the compound lens, and can be removed from the holder at the time of positioning and attached to the holder at the time of measurement. In the present invention, since the positioning element is located on the optical path of the direct light, if the spread of the direct light is smaller than the positioning element, the lens element is not directly irradiated with the direct light and Since the scattered light is not irradiated, the light shielding member is not necessarily required even if the light intensity of the direct light is large.

Further, the compound lens according to the present invention includes:
For example, a plurality of lens elements whose main axes are shifted in parallel can be arranged concentrically and integrated (claim 3).
Further, it is more preferable to provide a positioning element at a central portion where the plurality of lens elements are arranged concentrically (claim 4). Further, it is more preferable to cover a boundary portion between the lens element and the positioning element with a mask material (claim 5).

A plurality of lens elements having different focal lengths may be arranged concentrically, a positioning element may be provided at the center, and a boundary between the lens element and the positioning element may be covered with a mask material. (Claim 6).

Then, based on any of the above inventions,
The boundary between the plurality of lens elements may be covered with a mask material. Further, a plurality of lens elements having different focal lengths may be arranged concentrically, a positioning element may be provided at a central portion, and a boundary between the plurality of lens elements may be covered with a mask material.

Further, the positioning element can be realized by a convex lens (Claim 9), a pinhole (Claim 10) and the like. Furthermore, the lens element can be realized by a Fresnel lens (claim 11).

[0029]

FIG. 1 shows an example of a compound lens 15 according to the present invention. As shown in the figure, in this example, a convex lens 15a as a positioning element is provided at the center, and the first and second concentric circles on the two rings are arranged around the convex lens 15a.
The composite lens 15 is formed by joining and integrating the lens elements 15b and 15c in a state where they are positioned.

In this embodiment, both lens elements 15b, 15c
Are formed by cutting Fresnel lenses into appropriate shapes. Then, the composite lens 15 is formed by fitting the convex lens 15a and the two lens elements 15b and 15c together and bonding and bonding the bonding surfaces. Further, the convex lens 15a and the first
The second lens elements 15b and 15c have different focal positions. As a result, the light emitted from the same object point is collected by the convex lens 15a and the first and second lens elements 15b and 15c, but the image formation positions are different. As a specific structure for changing the focal position, the lens elements 15a to 15c can be realized by using different focal lengths or by changing the main axes.

FIG. 2 shows an embodiment relating to forward scattered light of a particle analyzer (flow cytometer) according to the present invention using the above-described compound lens. As shown in the figure, a flow cell 2 and a compound lens 15 are arranged in this order on the optical path of the light emitted from the laser light source 1. In addition,
Although not shown, a condensing lens is arranged on the emission side of the laser light source 1, and converges at the installation position of the flow cell 2. When the particles in the flow cell 2 are irradiated with laser light, forward scattered light is generated. And
The center (optical axis) of the path of the forward scattered light and the composite lens 15
Match with the center. Moreover, the optical axis and the composite lens 15 are arranged so as to be orthogonal to each other.

Further, each of the components 15a to 15c constituting the compound lens 15 has a convex lens 15 having a central focal length.
a is the shortest, and the outermost second lens element 15c is the longest. However, the principal axes of the elements 15a to 15c are made to coincide. As a result, light emitted from the same object point is condensed by passing through each of the elements 15a to 15c, but the imaging position is different. More specifically, all the images are formed on the center line (optical axis) of the compound lens 15, but the image forming position of the convex lens element 15a is closest to the compound lens 15 side, and the second lens element 15c is The image is formed at the position farthest from the camera.

On the other hand, in front of the compound lens 15, a first detector 16, a second detector 17, and a third detector 18 are arranged in this order. At this time, the centers of the detectors 16 to 18 and the compound lens 15 are located on the same line.
That is, the detectors 16 to 18 are arranged on the central axis of the optical path of the light emitted from the laser light source 1 so that the optical axes of the light receiving surfaces of the detectors 16 to 18 coincide with each other. Then, the first detector 16 is arranged at an image forming position of light that has passed through the convex lens element 15a having the shortest focal length. The second detector 17 is disposed at an image forming position of light that has passed through the first lens element 15b having an intermediate focal length. The third detector 18 has a second longest focal length.
It is arranged at the image forming position of the light passing through the lens element 15c.

Further, each of the detectors 16 to 18 is constituted by a photodiode (PD) and other photoelectric conversion elements. The outputs of the detectors 16 to 18 are supplied to the analyzer 9.
Where the analysis of the particles takes place. Further, the first detector 16 is provided with a pinhole so as to coincide with the center of the light receiving surface, so that light deviated from the center is not detected. Further, an analysis result based on the outputs of the second and third detectors 17 and 18 is output to the output device 19. Note that the analysis / classification processing performed based on signals from different scattering angles can be performed in the same manner as in the related art, and a detailed description thereof will be omitted.

Next, a measurement procedure using the above-described flow cytometer will be described. Prior to the measurement, there are positioning processing for adjusting the attitude of the compound lens 15 and actual analysis / classification processing performed after the positioning processing is completed. The positioning process may be performed each time the measurement is performed. Also,
The adjustment may be performed periodically or irregularly at a rate of once every several times.

* Positioning Processing Positioning processing is performed without supplying a sample serving as a scattering source to the flow cell 2. Since there is no scattering source, the laser light emitted from the laser light source 1 passes through the flow cell 2 as it is, and most of the laser light is applied to the convex lens 15a, which is the central positioning element of the composite lens 15, and forms an image at a predetermined position. If the composite lens 15 is located at a position shifted in the front-rear direction with respect to the flow cell 2 and the laser light source 1, no image is formed on the light receiving surface of the first detector 16. Also,
Even if there is a position in the front-back direction, the light
If the light is not irradiated to the center of the (convex lens 15a), an image is formed at the installation position of the first detector 16, but the composite lens 15
An image is formed at a position shifted from the center of the light receiving surface of the first detector 16 in accordance with the direction and distance of the shift. Then, in the present embodiment, a pinhole is provided on the front surface of the light receiving surface of the first detector 16 and only light that matches the central axis can pass therethrough. Cannot pass through the pinhole and does not reach the light receiving surface at the first detector 16. Similarly, light is emitted from the compound lens 15 (convex lens 15).
Even if the light is applied to the center of a), if the compound lens 15 is inclined, the light is applied obliquely to the center of the convex lens 15a, where it is refracted and the first detector 16
Is formed at a position deviated from the center of the light receiving surface. Therefore, the light is blocked by the pinhole and is not irradiated on the light receiving surface of the first detector 16.

Therefore, when the complex lens 15 is installed so that its center is on the optical axis and is orthogonal to the composite lens 15, and if it is installed so that the position in the front-back direction is in a desired state, the convex lens The light formed at 15a is applied to the light receiving surface of the first detector 16. As a result, the first detector 16
The posture of the compound lens 15 is adjusted while monitoring the output of. When the output of the first detector 16 is ON (light is received), it means that the complex lens 15 has been installed in a correct state.
The positioning processing ends.

Although the diameter of the convex lens 15a is shown relatively large for the convenience of writing the drawing, in order to perform the positioning process, it is necessary to determine whether or not light forms an image on the central axis of the first detector 16. Since it is only necessary to be able to judge this, the diameter of the convex lens 15a can also be reduced. Then, in the actual measurement to be described later, the light irradiated to the convex lens 15a is not used except for the case where the absorbance of the sample is measured, so that the convex lens 15a is also used from the viewpoint of effective use of the light at the time of measurement.
The smaller the diameter of a is, the better.

Since the composite lens 15 is actually fixed to a holder or the like installed at a predetermined position in the flow cytometer, the composite lens 15 has an approximate position and posture, and has at least one direction (for example, Positioning in the front-rear direction with respect to the optical axis) may be completed.
In such a case, adjustment in the remaining direction is performed.

In the above-described embodiment, an example in which a photodiode having a pinhole is used as the first detector 16 has been described. However, the present invention is not limited to this.
For example, a position detection photodiode (PSD: posi)
Whether or not an image is formed at the center of the light receiving surface may be determined by using, for example, a time sensitive detector.

In this embodiment, since the positioning can be performed without supplying the sample, the positioning is performed without supplying the sample. However, the positioning may be performed while supplying the sample. In addition, using the method described above,
Since there is no scattering, more light can be used for positioning, which is preferable.

* Analysis and Classification Processing The analysis processing is performed using the outputs of the second and third detectors 17 and 18. In other words, as is clear from the figure,
The forward small-angle scattered light having a small scattering angle (for example, 1.5 to 10 degrees) is applied to the first lens element 15b, and is condensed there to the second detector 17 located second from the complex lens 15. Received. Further, the forward large-angle scattered light having a large scattering angle (for example, 10 to 15 degrees) is applied to the outermost second lens element 15c, where it is collected and transmitted to the third detector 18 farthest from the compound lens 15. Received.

As described above, a single compound lens 15 can separate and condense a plurality of regions having different scattering angles. That is, in the related art, an optical system including a mirror with a hole, a collimator lens, and a plurality of condenser lenses can be configured with one compound lens, and the configuration is simplified, The occupation area is small,
Moreover, the adjustment becomes easy. Moreover, even if the number of regions to be separated increases, only the number of lens elements needs to be increased.
There is no increase in size. Furthermore, since only the light is condensed by the lens, almost all of the forward scattered light can be received by each detector, the light utilization rate is high, and even minute light can be reliably detected. .

FIG. 3 shows a second embodiment of the present invention in the case of forward scattered light. In the present embodiment,
Different from the first embodiment described above, each element 15a to 15a
15c has a different main axis. Thereby, each element 1
Light condensed by passing through 5a to 15c is imaged at a predetermined position on each main axis, and thus can be separated for each region having different scattering angles.

Of course, also in this case, the center of the compound lens 15 is a convex lens 15a as a positioning element, and the main axis A
Coincides with the optical axis of the light emitted from the laser light source 1, and the first detector 16 is installed on an extension of the optical axis.

Further, the forward small-angle scattered light having a small scattering angle is radiated to the first lens element 15b, where it is condensed and the main axis B is shifted downward with respect to the main axis A in the figure.
The light is received by the second detector 17 arranged at the upper predetermined position. Further, the forward large-angle scattered light having a large scattering angle is radiated to the outermost second lens element 15c, where it is condensed and the main axis C is displaced upward with respect to the main axis A in the figure.
The light is received by the third detector 18 disposed at the upper predetermined position.

Although not shown, each of the detectors 16 to 1
The output of 8 is given to the analyzer as in FIG. Then, the positioning of the compound lens 15 is performed based on the output of the first detector 16. In addition, the second and third detectors 17, 1
The analysis and classification of the particles in the flow cell 2 are performed based on the output of 8.

In the above example, each of the elements 15a to 15a
Since the focal lengths 5c are equal, the detectors 16 to 18 are installed at the same distance from the compound lens 15 as shown in the figure, but the present invention is not limited to this. Of course, the distances may be different. Note that the other configuration and operation and effect are the same as those of the above-described first embodiment, and a detailed description thereof will be omitted.

FIG. 4 shows an embodiment in which the present invention is applied to a flow cytometer using side scattered light and back scattered light in addition to forward scattered light. In FIG. 1, irradiation light a emitted from a laser light source 1 passes through a central portion of a compound lens for separating back scattered light, irradiates particles 7, which are scattering sources of a flow cell 2, and forward scattered light b, c, d,
Side scattered light e, f, g and back scattered light h, i are obtained. Each scattered light is separated by the compound lens 15 arranged one by one in each direction, and the intensity of light is detected by each detector 6. The direct light passing through the flow cell 2 from the laser light source 1 is configured to be detected by a detector 6a for positioning. The present embodiment is an example in which the complex lens 15 of the present invention is applied to the conventional example of the flow cytometer shown in FIGS. In the conventional example, as the number of scattered lights to be detected increases, the number of optical systems 8 including the collimator lens 3, the mirror 4, and the condenser lens 5 increases, which is a factor that hinders miniaturization of the apparatus. If the compound lens 15 of the present invention is used, the optical system 8 can be replaced with one compound lens, and a small and inexpensive flow cytometer can be supplied. Further, conventionally, it was necessary to adjust the optical axis of each of the collimator lens 3, the mirror 4, and the condenser lens 5. However, if only the composite lens 15 is adjusted, the measurement using the apparatus becomes extremely easy. Has merits. Further, when the compound lens is manufactured by a mold, it is possible to essentially eliminate the variation of the focal position for each lens element in one compound lens.

FIG. 5 shows an example of a method of manufacturing the compound lens 15 used in the second embodiment.
In the second embodiment, the first and second lens elements 15b,
The focal lengths of the lenses 15c are the same, and only the main axis is different. Therefore, as shown by a two-dot chain line in the figure, a mother lens 20 composed of a large Fresnel lens is prepared, and the mother lens 20 is cut out at a position shifted from the main axis of the mother lens 20 by a predetermined distance to form a ring having a medium diameter. The lens element 15b is formed. Since the main axis of the first lens element 15b is the main axis of the mother lens 20, it is shifted to the left side in the figure with respect to the center O1 of the ring. The second lens element 15c is formed by punching out a large-diameter ring at a position shifted by a predetermined distance from the main axis of the mother lens 20 to the side opposite to the first lens element 15b. This second lens element 15c
Is the main axis of the mother lens 20, and thus is shifted to the right in the figure with respect to the center O2 of the ring. Therefore,
The principal axes of the first lens element 15b and the second lens element 15c are located at positions separated from the center by a predetermined distance, and the directions of displacement of the two lens elements 15b and 15c are reversed.
A plurality of lens elements can be manufactured by appropriately punching out one large Fresnel lens as described above, and a composite lens can be easily manufactured by joining them. Such a manufacturing process becomes more effective in producing lens elements having the same focal length as the number of lens elements to be joined increases. Needless to say, it is not always necessary to manufacture each lens element from one Fresnel lens.

That is, since this lens is intended to have a function of focusing light irradiated on a plurality of regions having a predetermined area for each of the regions, it is suitable for the design of a device such as a Fresnel lens. A plurality of lenses having a focal length are prepared, and a plurality of lenses are cut or cut out from the lenses so as to have a predetermined shape and area to form lens elements, and the lens elements are integrally formed into a planar shape. It can be manufactured by joining.
The focal lengths of the plurality of lenses prepared may be different from each other, and the principal axes of the lens elements may be designed to be coincident or shifted. By such molding, the optical system unit can be manufactured compactly in a limited space.

As a method of manufacturing the compound lens 15, in addition to the above-described method, a surface of a mold is prepared in advance by integrating each lens element or by combining them.
Using the mold, it can be manufactured by a technique such as compression molding, ICM molding (injection compression molding), or injection molding. An example of the production will be described below with reference to FIGS.

That is, the mold A ′ is molded by cutting or hollowing out the mold A from which the Fresnel lens A can be molded (see FIG. 6). Further, an annular mold B ′ provided with a hole capable of incorporating the mold A ′ is replaced with a mold B capable of molding the Fresnel lens B.
And cut out or cut out from it (see FIG. 7). Further, the mold surface can be formed by incorporating the mold A ′ into the mold B ′ (see FIG. 8). The mold A and the mold B may be the same, or may be molds for molding Fresnel lenses having different focal lengths or different main axes.

FIG. 9 shows another embodiment of the present invention.
This is an improvement of the compound lens 15. That is,
Also in this embodiment, 2 of the first and second lens elements 15b and 15c
The two areas are separated and detected. Then, the joining region between the adjacent lens elements and the joining region between the convex lens 15a and the first lens element 15b are formed into a ring-shaped mask material 2.
It is covered with 2. The mask member 22 is formed by joining the components 15a to 15c, and then attaching an optically opaque material to the joint. Specifically, it can be realized by patterning after depositing and sputtering a predetermined thin film, applying predetermined ink or the like, or attaching a tape material or the like.

That is, if the mask material 22 is not provided, light may be irregularly reflected at the joint portion and may not be correctly focused on a predetermined detector, and may become a noise component. However, by providing the mask material 22 as in the present embodiment, light passes only through the normal lens element portion, so that the detector can receive light in a desired state. And
The compound lens 15 provided with such a mask member 22 can be applied to any of the embodiments described above.

In each of the embodiments described above, the example in which the convex lens 15a is used as the positioning element has been described.
The present invention is not limited to this.
A pinhole may be provided at the center of the frame, and the pinhole may be used as a positioning element.

That is, if a pinhole is formed, for example, if the center of the compound lens 15 does not coincide with the optical axis of the light emitted from the laser light source 1, the light cannot pass through the pinhole and the first detector receives the light. Not illuminated on the surface. Further, when the compound lens 15 is disposed obliquely, the amount of light passing through the pinhole decreases, and when the output of the first detector 16 decreases or the inclination angle increases, the composite lens 15 can pass through the pinhole. Disappears. Therefore, the posture of the compound lens 15 can be controlled by monitoring the output of the first detector.
Specifically, it can be determined that the positioning has been performed when the light amount becomes equal to or more than a certain value or when the light amount becomes maximum. However, when this pinhole is used, it is not possible to cope with a positional shift in the front-back direction along the optical axis direction. Therefore, when the positioning in the front-rear direction is unnecessary, there is an advantage that the lens can be formed with a simple configuration such as providing a pinhole at the center of the compound lens.

In each of the above embodiments, the compound lens is formed by combining two lens elements, but three or more lens elements may be used. Further, in each of the embodiments described above, the lens element, and thus the compound lens, is configured using a Fresnel lens, but the present invention is not limited to this, and can be configured using a normal lens such as a convex lens. .

Further, although not shown, in each of the above-described embodiments, an obsculator (having the same function as the reference numeral 11 shown in FIG. 15) as a light shielding member is provided at a predetermined position between the flow cell and the compound lens. May be installed, and the direct light emitted from the laser light source 1 may be shielded there. However, the obscurator is movable, and is located on the optical path of direct light during normal measurement, but when adjusting the posture of the compound lens,
It is retracted from the optical path so that direct light from the laser light source 1 can enter the positioning element 15a. Also,
The light-shielding member is not limited to a conventional obsculator, but may be a member that is directly or indirectly attached to the compound lens 15, and its structure is arbitrary.

In contrast to the above-described embodiment, even during the actual measurement after the attitude of the compound lens is adjusted, the light between the flow cell and the compound lens can be appropriately adjusted without directly blocking the light with the obsculator. If a condenser lens is arranged at the position and the output of the first detector 16 is detected, the absorbance of the particles flowing in the flow cell can be measured.

FIG. 10 shows another embodiment of the compound lens 35 according to the present invention. The difference from the embodiment shown in FIG. 1 is that no positioning mechanism is provided. And, specifically, it is as follows. As shown in the figure,
In this example, three lens elements 35a, 35 are concentrically formed.
The composite lens 35 is formed by joining and integrating the members while b and 35c are positioned. That is, the first lens element 35a at the center is disk-shaped, and two ring-shaped second and third lens elements 35b, 35 are formed around the first lens element 35a.
c is located.

In this example, each of the lens elements 35a to 35c
Are formed by cutting Fresnel lenses into appropriate shapes. Then, the lens elements 35a to 35c are fitted together, and the bonding surfaces are bonded and integrated to form one composite lens 35.
Further, the first to third lens elements 35a to 35c have different focal positions. Thereby, the light emitted from the same object point is transmitted to each of the lens elements 35a to 35a.
The light is condensed at 5c, but the image formation position is different. A specific structure for changing the focal position can be realized by using each of the lens elements 35a to 35c having a different focal length or having a different main axis.

FIG. 11 shows a fourth example of the particle analyzer (flow cytometer) according to the present invention using the above-described compound lens.
Is shown. As shown in the figure, a flow cell 2 and a compound lens 15 are arranged in this order on the optical path of the light emitted from the laser light source 1. When the particles in the flow cell 2 are irradiated with laser light, forward scattered light is generated. Then, the center of the path of the forward scattered light and the center of the compound lens 35 are matched.

Further, each of the lens elements 35a to 35c constituting the compound lens 35 has the shortest first lens element 35a having a central focal length and the third lens element 3
5c is the longest. However, the principal axes of the lens elements 35a to 35c are made to coincide. As a result, light emitted from the same object point is
The light is condensed by passing through a to 35c, but its imaging position is different. More specifically, each of the composite lenses 35
Is formed on the center line of the first lens element 35a, the image forming position of the first lens element 35a is closest to the compound lens 35 side, and the third lens element 35c is formed at the position farthest from the compound lens 35.

On the other hand, a first detector 36, a second detector 37, and a third detector 38 are arranged in front of the compound lens 35 in this order. At this time, the centers of the detectors 36 to 38 and the compound lens 35 are located on the same line. Then, the first detector 36 is disposed at an image forming position of light that has passed through the first lens element 35a having the shortest focal length. The second detector 37 is disposed at an image forming position of light that has passed through the second lens element 35b having an intermediate focal length. The third detector 38 is a third lens element 35c having the longest focal length.
Is arranged at the image forming position of the light that has passed through.

Further, each of the detectors 36 to 38 is constituted by a photoelectric conversion element. Then, the output of each of the detectors 36 to 38 is given to the analyzer 9, where the particles are analyzed. Then, the analysis result is output to the output device 39. Note that the analysis / classification processing performed based on signals from different scattering angles can be performed in the same manner as in the related art, and a detailed description thereof will be omitted.

In the above-described configuration, as is apparent from the drawing, the forward small-angle scattered light having a small scattering angle is radiated to the central first lens element 35a, where it is condensed and the first small-angle scattered light is closest to the compound lens 35. The light is received by one detector 36.
Further, the forward mid-angle scattered light having a middle scattering angle is applied to the second lens element 35b, collected there, and received by the second detector 37 located second from the compound lens 35. Further, the forward large-angle scattered light having a large scattering angle is radiated to the outermost third lens element 35c, collected there, and received by the third detector 38 farthest from the compound lens 35.

As described above, a plurality of regions having different scattering angles can be separated and condensed by one compound lens 35. That is, in the related art, an optical system including a mirror with a hole, a collimator lens, and a plurality of condenser lenses can be configured with one compound lens, and the configuration is simplified, The occupation area is small,
Moreover, the adjustment becomes easy. Moreover, even if the number of regions to be separated increases, only the number of lens elements needs to be increased.
There is no increase in size. Furthermore, since only the light is condensed by the lens, almost all of the forward scattered light can be received by each detector, the light utilization rate is high, and even minute light can be reliably detected. .

FIG. 12 shows a fifth embodiment of the present invention. In the present embodiment, unlike the above-described fourth embodiment, the principal axes of the lens elements 35a to 35c are different. Thereby, each lens element 35a-35
Light condensed after passing through c is imaged at a predetermined position on each principal axis, so that the light can be separated into regions having different scattering angles.

Specifically, the forward small-angle scattered light having a small scattering angle is applied to the central first lens element 35a, where it is collected and focused on the first detector 36 disposed at a predetermined position on the main axis A. Received. Further, middle forward scattered light having a middle scattering angle is applied to the second lens element 35b, collected there, and arranged at a predetermined position on the main axis B, which is displaced downward with respect to the main axis A in the figure. The received second detector 37 receives the light. Further, the forward large-angle scattered light having a large scattering angle is applied to the third lens element 35c on the outermost periphery, is condensed there, and is disposed at a predetermined position on the main axis C which is displaced upward with respect to the main axis A in the figure. The received third detector 38 receives the light.

Although not shown, each of the detectors 36 to 3
The output of 8 is given to the analyzer in the same manner as in FIG. 11, and the particles in the flow cell 2 are separated. In the above example, since the focal lengths of the lens elements 35a to 35c are equal, the detectors 36 to 38, as illustrated,
Although installed at a position separated by an equal distance from the compound lens 35, the present invention is not limited to this, and it is needless to say that the focal length may be different. Note that the other configuration and operation and effect are the same as those of the above-described fourth embodiment, and a detailed description thereof will be omitted.

FIG. 13 shows an example of a method of manufacturing the compound lens 35 used in the fifth embodiment. In the fifth embodiment, each of the lens elements 35a to 35a
The focal lengths of c are equal, and only the main axis is different. Therefore, as shown by a two-dot chain line in the figure, a first lens element 35a is formed by preparing a mother lens 40 composed of a large Fresnel lens and punching it out into a small disk shape around the main axis of the mother lens 40. . The second lens element 35b is formed by punching out a medium-diameter ring at a position shifted from the main axis of the mother lens 40 by a predetermined distance. The main axis of the second lens element 35b is
, The position is shifted leftward in the figure with respect to the center O1 of the ring. The third lens element 35c is formed by punching out a large-diameter ring at a position shifted by a predetermined distance from the main axis of the mother lens 40 to the side opposite to the second lens element 35b. Since the main axis of the third lens element 35c is the main axis of the mother lens 40, the center O2 of the ring
Is shifted to the right in the figure. Therefore, the main axis of the first lens element 35a is located at the center of the disk, and
The principal axes of the lens element 35b and the third lens element 35c are located at positions separated from the center by a predetermined distance, and the directions of displacement of the two lens elements 35b and 35c are reversed. A plurality of lens elements can be manufactured by appropriately punching out one large Fresnel lens as described above, and a composite lens can be easily manufactured by joining them. Needless to say, it is not always necessary to manufacture each lens element from one Fresnel lens.

FIG. 14 shows a sixth embodiment of the present invention, in which the composite lens 35 is improved. That is, also in this example, three areas of the first to third lens elements 35a to 35c are separately detected. Then, the bonding region between the adjacent lens elements is formed into a ring-shaped mask material 42.
To cover it. The mask material 42 is formed by joining the lens elements 35a to 35c and then attaching an optically opaque material to the joint. Specifically, it can be realized by patterning after depositing and sputtering a predetermined thin film, applying predetermined ink or the like, or attaching a tape material or the like. In this case, at the joint portion, light may not be collected properly on a predetermined detector due to irregular reflection or the like, and may become a noise component. However, by providing the mask material 42, only the normal lens element portion is provided. Is transmitted, so that the detector can receive light in a desired state. The composite lens 35 provided with the mask member 42 can be applied to any of the above-described embodiments.

In each of the above embodiments, the compound lens is formed by combining three lens elements. However, two or four or more lens elements may be used. Further, in each of the embodiments described above, the lens element, and thus the compound lens, is configured using a Fresnel lens, but the present invention is not limited to this, and can be configured using a normal lens such as a convex lens. .

[0075]

As described above in detail, according to the present invention, a plurality of lens elements having different focal positions are joined and integrated to produce one compound lens, and the light having different scattering angles is separated by the compound lens. And can be assigned to each detector. In addition, since one compound lens can be used alone, the number of components can be reduced and the optical system can be easily adjusted. Furthermore, since it is only focused by the lens,
Light utilization also increases. Furthermore, even if the number of regions to be divided is increased, only the number of lens elements is increased, and the number of parts is one.
A single compound lens may be used, and the number of components of the optical system for separating light does not increase, and the occupied area does not increase, so that the space due to the increase in detectors can be limited.

In addition, when a positioning element such as a convex lens or a pinhole is provided at the center of the compound lens, the correct position and posture of the compound lens is determined depending on whether or not light normally passes through the positioning element. Can be determined, and the positioning process can be performed easily and accurately.

[Brief description of the drawings]

FIG. 1 is a diagram showing an example of a compound lens according to the present invention.

FIG. 2 is a diagram showing a first embodiment of a particle analyzer according to the present invention.

FIG. 3 is a view showing a second embodiment of the particle analyzer according to the present invention.

FIG. 4 is a diagram showing a third embodiment of the particle analyzer according to the present invention.

FIG. 5 is a diagram illustrating an example of a method of manufacturing a compound lens.

FIG. 6 is a diagram (part 1) illustrating another example of the method for manufacturing a compound lens.

FIG. 7 is a diagram (part 2) illustrating another example of the method for manufacturing a compound lens.

FIG. 8 is a diagram (part 3) illustrating another example of the method for manufacturing a compound lens.

FIG. 9 is a diagram showing another example of the compound lens according to the present invention.

FIG. 10 is a diagram showing an example of a compound lens according to the present invention.

FIG. 12 is a view showing a fourth embodiment of the particle analyzer according to the present invention.

FIG. 13 is a view showing a fifth embodiment of the particle analyzer according to the present invention.

FIG. 14 is a diagram illustrating an example of a method for manufacturing a compound lens.

FIG. 15 is a diagram showing a conventional example.

FIG. 16 is a diagram showing another conventional example.

FIG. 17 is a diagram showing another conventional example.

FIG. 18 is a diagram showing another conventional example.

[Explanation of symbols]

 6, 6 'detector 9 analyzer 15 compound lens 15a convex lens element 15b first lens element 15c second lens element 16 first detector 17 second detector 18 third detector 22 mask material 35 composite lens 35a first lens Element 35b Second lens element 35c Third lens element 36 First detector 37 Second detector 38 Third detector 42 Mask material

[Procedure amendment]

[Submission date] May 27, 1998

[Procedure amendment 1]

[Document name to be amended] Statement

[Correction target item name] Brief description of drawings

[Correction method] Change

[Correction contents]

[Brief description of the drawings]

FIG. 1 is a diagram showing an example of a compound lens according to the present invention.

FIG. 2 is a diagram showing a first embodiment of a particle analyzer according to the present invention.

FIG. 3 is a view showing a second embodiment of the particle analyzer according to the present invention.

FIG. 4 is a diagram showing a third embodiment of the particle analyzer according to the present invention.

FIG. 5 is a diagram illustrating an example of a method of manufacturing a compound lens.

FIG. 6 is a diagram (part 1) illustrating another example of the method for manufacturing a compound lens.

FIG. 7 is a diagram (part 2) illustrating another example of the method for manufacturing a compound lens.

FIG. 8 is a diagram (part 3) illustrating another example of the method for manufacturing a compound lens.

FIG. 9 is a diagram showing another example of the compound lens according to the present invention.

FIG. 10 is a diagram showing an example of a compound lens according to the present invention.

FIG. 11 is a diagram showing a fourth embodiment of the particle analyzer according to the present invention.

FIG. 12 is a diagram showing a fifth embodiment of the particle analyzer according to the present invention.

FIG. 13 is a diagram illustrating an example of a method for manufacturing a compound lens.

FIG. 14 is a diagram showing a main part of a particle analyzer according to a sixth embodiment of the present invention.

FIG. 15 is a diagram showing a conventional example.

FIG. 16 is a diagram showing another conventional example.

FIG. 17 is a diagram showing another conventional example.

FIG. 18 is a diagram showing another conventional example.

[Explanation of Signs] 6, 6 'detector 9 analyzer 15 compound lens 15a convex lens element 15b first lens element 15c second lens element 16 first detector 17 second detector 18 third detector 22 mask material 35 composite Lens 35a First lens element 35b Second lens element 35c Third lens element 36 First detector 37 Second detector 38 Third detector 42 Mask material

 ──────────────────────────────────────────────────続 き Continuing on the front page (72) Katsuhiro Tsuchiya, Inventor, Katsuhiro Tsuchiya 1-31-4, Nishi-Ochiai, Shinjuku-ku, Tokyo Japan Photovoltaic Industry Co., Ltd. (72) Yoshiyuki Takahara, 1-31-1, Nishi-Ochiai, Shinjuku-ku, Tokyo No. Japan photoelectric company

Claims (11)

[Claims] 1. A particle analyzer which classifies the particles by separating and detecting those having different scattering angles among scattered lights generated by irradiating the particles with light, wherein the scattered light is A plurality of ring-shaped lens elements having different focal positions, which are arranged so as to block the optical path, are integrated in a state where they are located on concentric circles; and A detector for detecting each scattering angle arranged at an image forming position of the scattered light, and an analysis unit for classifying the particles based on detection outputs of the plurality of detectors, Particle analyzer. 2. A particle analyzer which classifies the particles by separating and detecting those having different scattering angles among scattered lights generated by irradiating the particles with light, wherein the scattered light is A plurality of ring-shaped lens elements having different focal positions, which are arranged so as to block the optical path, are located on concentric circles, and a compound lens integrated with a positioning element provided in a central portion; A detector for detecting each scattering angle arranged at an image forming position of the scattered light according to the focal position of the lens element; a positioning detector capable of detecting light passing through the positioning element; Analyzing means for classifying the particles based on the detection output of the detector, and wherein the positioning element has at least the center of the compound lens and the optical axis coincide with each other. When the compound lens is orthogonal to the optical axis, a predetermined amount of light can be received at a predetermined position of the positioning detector installed in front. Particle analyzer. 3. A compound lens wherein a plurality of lens elements whose principal axes are shifted in parallel are concentrically arranged and integrated. 4. The compound lens according to claim 3, wherein a positioning element is provided at a central portion where the plurality of lens elements are arranged concentrically. 5. The compound lens according to claim 3, wherein a boundary between the lens element and the positioning element is covered with a mask material. 6. A plurality of lens elements having different focal lengths are arranged concentrically, a positioning element is provided at a central portion, and a boundary between the lens element and the positioning element is covered with a mask material. Complex lens. 7. The compound lens according to claim 3, wherein a boundary between the plurality of lens elements is covered with a mask material. 8. A composite, wherein a plurality of lens elements having different focal lengths are arranged concentrically, a positioning element is provided at a central portion, and a boundary portion between the plurality of lens elements is covered with a mask material. lens. 9. The compound lens according to claim 4, wherein the positioning element is a convex lens. 10. The compound lens according to claim 4, wherein said positioning element is a pinhole. 11. The compound lens according to claim 3, wherein the lens element is a Fresnel lens.