CN1318847C - Electrophoresis apparatus - Google Patents
Electrophoresis apparatus Download PDFInfo
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- CN1318847C CN1318847C CNB951011944A CN95101194A CN1318847C CN 1318847 C CN1318847 C CN 1318847C CN B951011944 A CNB951011944 A CN B951011944A CN 95101194 A CN95101194 A CN 95101194A CN 1318847 C CN1318847 C CN 1318847C
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- electrophoresis equipment
- migrating channels
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44756—Apparatus specially adapted therefor
- G01N27/44782—Apparatus specially adapted therefor of a plurality of samples
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- G—PHYSICS
- G01—MEASURING; TESTING
- G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
- G01N27/00—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means
- G01N27/26—Investigating or analysing materials by the use of electric, electrochemical, or magnetic means by investigating electrochemical variables; by using electrolysis or electrophoresis
- G01N27/416—Systems
- G01N27/447—Systems using electrophoresis
- G01N27/44704—Details; Accessories
- G01N27/44717—Arrangements for investigating the separated zones, e.g. localising zones
- G01N27/44721—Arrangements for investigating the separated zones, e.g. localising zones by optical means
Abstract
An electrophoresis apparatus for detecting samples migrating in migration portions includes a plurality of gel separation portions separate from one another and having respective ends disposed along a straight line, a plurality of migration portions equal in number to the gel separation portions and disposed along a straight line following respective ones of the gel separation portions, a light source for generating light which passes through the migration portions, the light propagating along a straight line extending through all of the migration portions, a plurality of optical fibers having respective first ends and respective second ends, the first ends of the optical fibers being disposed facing respective points where the light from the light source passes through respective ones of the migration portions, the optical fibers being equal in number to or greater in number than the migration portions, and an optical detector optically coupled to the second ends of the optical fibers for receiving light from the migration portions via the optical fibers.
Description
The present invention relates to be used for separating and analytical equipment of DNA and other biological substance, and be particularly related to electrophoresis equipment.
In prior art, gel electrophoresis has been used to comprise the DNA analysis of DNA (DNA (deoxyribonucleic acid)) order-checking.Requirement to dna sequencing in recent years constantly increases, as in genome analysis.Simultaneously, can obtain a kind of dna sequencing instrument on market, this dna sequencing instrument is measured dna fragmentation by utilizing the fluorescent label DNA fragment on real-time basis when carrying out the gel electrophoresis separation; This equipment is applied in practice.This dna sequencing instrument uses tabular gel (polyacrylamide gel), and it provides 30 to 40 migrating channels.To laser beam irradiation, and sequenator detects by irradiation place the time emitted fluorescence with fluorescently-labeled dna fragmentation institute to this gel slab from electrophoresis starting point gap in accordance with regulations.
But the throughput of this dna sequencing instrument available on the market is very little, for every day the 10K base to the 20K base; This is one that hinders in the problem of genome analysis.In order to obtain high throughput, improving electrophoretic velocity and very many migrating channels are provided is effectively, can increase the quantity of the sample that the same time detects like this.
Developed a kind of capillary gel electrophoresis device that can realize electrophoretic separation at short notice in recent years.And then, also develop a kind of capillary array electrophoresis device, it allows to measure simultaneously under a large amount of structures capillaceous, thereby reaches high throughput.
Reported the capillary array electrophoresis device of following described type: a kind of its capillary array that is banded in the flat board that installs is fixed on the X-Y platform, and capillary array exposes laser beam from the top so that detect fluorescence (R.A.Mathies etc. according to the direction of laser beam, 359 phases of Nature (Nature Journal), 167 to 169 pages (1992)).In described device, in synchronization, have only a kapillary that laser beam is exposed, and speed is carried out mechanical scanning so that all kapillaries are measured fluorescence signal to capillary gel in accordance with regulations.
This common capillary array electrophoresis device in 359 phases of Nature (Nature Journal), 167 to 169 pages (1992) publication can improve throughput significantly, but the maximum quantity of migrating channels is 20 to 40.This is because if increase quantity capillaceous, will reduce each minute capillaceous, and cause sensitivity low.Because the quick scanning to radiation is impossible, must reduce electrophoretic velocity.
Purpose of the present invention is exactly this problem that solves in the prior art, and a kind of electrophoresis equipment is provided.Although increase quantity capillaceous, this electrophoresis equipment can guarantee under the situation of not sacrificing sensitivity and electrophoretic velocity the detection to sample.
In order to reach this purpose, according to the present invention, by a kind of layer form arrangement kapillary more than two, and the end of the gel-filled kapillary sample of the usefulness of the migratory direction that in the light unit, is arranged along a straight line; Buffer solution is fed to the place of close gel-filled capillary end to form outer surface low (Sheath flow) then.Laser beam shines these positions simultaneously, on these positions with fluorescently-labeled DNA outside each gel-filled end capillaceous is eluted to the surface low, and flow downward.Optical fiber is disposed near illuminated and sentences collection by being directed to line sensor or zone sensors (dimension sensor) with fluorescently-labeled DNA institute's emitted fluorescence and fluorescence.
According to an aspect of the present invention, provide the electrophoresis equipment of a kind of DNA of detection or other biological substance, it comprises:
(1) many migrating channels that are used to handle the electrophoresis of DNA or other biological material, their end is arranged and is isolated mutually along straight line;
(2) light sources that produce light, the light of generation are along described straight-line pass, and are crossing with all migrating channels;
(3) optical receiving fiber, the end of optical receiving fiber be arranged in and the light of light source and corresponding optical axis that the intersection point between the described migrating channels is complementary on, and the quantity of optical receiving fiber is equal to or greater than the quantity of migrating channels; And
(4) optical detection devices, its other end with each bar optical fiber is connected, and is used to receive light.
According to another aspect of the present invention, provide a kind of electrophoresis equipment that is used to detect DNA or other biological substance, it comprises:
(1) many migrating channels that are used to handle the electrophoresis of DNA or other biological material, their end is arranged and is isolated mutually along straight line;
(2) fluorescence detection units that hold an end of described migrating channels thereon;
(3) light sources that produce light, the light of generation are along described straight-line pass, and are crossing with all migrating channels;
(4) optical receiving fiber, the end of optical receiving fiber be arranged in and an end of the light of light source and migrating channels between the corresponding optical axis that is complementary of intersection point on, and the quantity of optical receiving fiber is equal to or greater than the quantity of migrating channels; And
(5) optical detection devices, its other end with each bar optical fiber is connected, and is used to receive light.
In a preferred embodiment, migrating channels comprises the gel capillary portion, and collects the optical fiber reception sample emitted fluorescence of light.
In a preferred embodiment, optical detection device comprises one-dimensional array sensor or two-dimensional array sensor, and the other end that is used for collecting the optical fiber of light optically is connected with a photo-sensitive cell of one-dimensional array sensor or two-dimensional array sensor at least.
In a preferred embodiment, ray-collecting lens are between an end of the optical fiber of described point of irradiation and reception light, and two or more bandpass filter is placed between the other end and optical detection device of optical fiber.
In a preferred embodiment, the optical fiber that forms the light collecting lens at the one end can be used as optical fiber.
In a preferred embodiment, this equipment has a kind of division source light, to shine the device of two or more fluoroscopic examination optical elements.
The present invention allows a laser beam to shine all positions near the kapillary termination simultaneously, and is irrelevant with quantity capillaceous, on these positions with fluorescently-labeled DNA by wash-out.
With to utilize lens to form the method for an images of two or more migration positions inequality, the fluorescence of the DNA that is labeled for detection, ray-collecting lens and optical fiber are placed in the junction of each migration position in the light receiving part; Like this, guaranteed effective detection to fluorescence, and irrelevant with quantity capillaceous.
Authors report of the present invention is crossed this capillary array electrophoresis equipment and is had following characteristics: buffer solution along the gel Capillary Flow in optical element, forming an outer surface low by the gel of straight line end capillaceous, thereby and side by side shine all migrating channels along described straight line emission laser beam near surface low district end outside being somebody's turn to do; So and the fluorescent image of acquisition is measured (H.Kambara etc., 361 phases of Nature (Nature Journal), 565 to 566 pages (1993) by a CCD camera or analog.
These lens of described equipment utilization focus on the fluorescent image that irradiation obtained by near the outer surface low part of detector end the time.When the length in this illuminated district was big, because the length of sensor array has only 24mm so little, the fluorescent image that is used to detect was weakened.Weakening of fluorescent image causes low phosphor collection efficient.Attempting increases the reduction that quantity capillaceous will cause phosphor collection efficient, thereby causes sensitivity to reduce, and therefore is difficult to the iridescent signal is separated from each bar migrating channels.
But,, be by realizing on the photo-sensitive cell that the output end of each optical fiber is seated in line sensor or face sensor to the detection of fluorescence according to the present invention; Like this permission can detect with the as many migrating channels of light receiving element quantity on the fluorescence that sends.According to this mode described above, might improve the detection sensitivity of (Nature Journal Nature361 phase, 565 to 566 pages (1993)) the sort of capillary array electrophoresis equipment of having reported of author of the present invention, thereby reach the performance of enhancing.
The face sensor that has the line sensor of about 1000 photosensitive units (light receiving element) and have 100,000 or a more photosensitive unit (light receiving element) can access.The invention provides a kind of electrophoresis equipment, it in fact uses the countless kapillary that is arranged to array, and making design a kind of is that the dna sequencing instrument of feature becomes possibility with very high throughput.
Fig. 1 is a synoptic diagram, and expression is according to the structure of the electrophoresis equipment in the first embodiment of the present invention;
Fig. 2 is illustrated in according to a kind of method of utilizing optical fiber that the detecting device of phosphor collection part and detection fluorescence is coupled together in the first embodiment of the present invention;
Fig. 3 is a synoptic diagram, is illustrated in the middle according to a second embodiment of the present invention structure that adopts the electrophoresis equipment of two or more fluorescence detection units; And
Fig. 4 is a synoptic diagram, is illustrated in the structure of the electrophoresis equipment in a third embodiment in accordance with the invention.
Below to describing in detail according to various embodiments of the present invention.
First embodiment
The following describes the situation that electrophoresis equipment according to the present invention is applied to dna sequencing:
In utilizing the dna sequencing of enzyme reaction, by adopting archaeal dna polymerase and target DNA being carried out synthetic four the dna fragmentation groups (fragment family) of generation of complementary strand as template DNA.In other words, complementary strand begins to synthesize from having the oligomer that can hybridize the special base sequence on target DNA, have the fragment of different length with generation, the end of these fragments has the analog of gland fat purine (A), thymine (T), cytimidine (C) or the black purine (G) that can not further extend.Its end is called A family, T family, G family and C family for the fragment of A, T, G and C analog.
Distinguishing by spectrographic technique, and dna fragmentation length is separated with gel electrophoresis with different fluorescence institute marks in these families.Four mixed each samples then of family utilize a capillary gel separated.From the shortest dna fragmentation, dna fragmentation passes through irradiation position in order.From can differentiate the type of the base of dna fragmentation end with fluorescently-labeled dna fragmentation institute emitted fluorescence wavelength.Realize dna sequencing according to described method.
In order to discern four types terminal bases, must detect two kinds or more kinds of fluorescence (the best is four kinds) in this case by selection to wavelength of fluorescence.And then the gel electrophoresis analysis of dna gel not only can be applicable to dna sequencing and also can be applicable to gene diagnosis, only uses one type fluorescence promptly enough in latter event.
Fig. 1 is a synoptic diagram, and capillary array equipment is described, it is according to an electrophoresis equipment of the present invention.The various sequencing reaction products that form by different DNA samples by electrophoresis be injected into the top of gel kapillary 1.The upper end of gel kapillary 1 is immersed in the specimen bottle that contains four fragment families, applies the voltage of about 10KV being immersed between electrode in the specimen bottle and this lower end capillaceous, so that dna fragmentation is injected in the kapillary.
After dna fragmentation is injected into kapillary 1; The upper end of gel kapillary 1 is tied and inserts in the buffer solution of 19 li of buffer containers that are deployed in the top position.Between two iontophoretic electrodes 17, apply 10 to 15KV voltage, one of them iontophoretic electrode 17 is immersed in the top buffer solution of 19 li of buffer containers that are deployed in the top position, another iontophoretic electrode is immersed in the buffer solution of 18 li of buffer containers that are deployed in lower position, arranged the lower end of uncovered kapillary 6 in lower position, so that buffer solution flows out from these uncovered kapillaries.Buffer solution mobile causes surface low outside capillaceous, and dna fragmentation will be moved in the outer surface low like this.
The length of gel-filled kapillary 1 is generally 20 to 40cm, but length can be 10 to 15cm when analyzing short DNA or when not requiring high score from performance.When analyzing length dna or when requiring high score, use 50 to 100cm long gel filled capillary pipe 1 possibly from performance.
Be the fluoroscopic examination to fragment, the lower part of gel-filled kapillary 1 is exposed to the laser beam 4 times that lasing light emitter 5 sends.If the kapillary body is directly exposed under the laser beam, light beam will scattering and refraction on the microcapillary tube surface, and the result can not be shone all kapillaries simultaneously.
In order to address this problem, the lower end of gel kapillary 1 is arranged to straight line and is immersed in the buffer solution of light unit 2, is radiated at simultaneously on the position near the lower end of all gel-filled kapillary 1 to guarantee laser beam 4 actually; Thereby the simultaneity that guarantees in fact dna fragmentation that the lower end wash-out from gel-filled kapillary 1 is gone out detects.In order to prevent that after dna fragmentation is by wash-out DNA is with 3 diffusion, the lower end that uncovered kapillary 6 is arranged in each gel-filled kapillary 1 is sentenced the end facing to each gel-filled kapillary 1, buffer solution flows into light unit 2 from the bottle 7 of putting buffer solution like this, and comes out to form gel-filled kapillary outer surface low on every side from uncovered capillary flow.
As shown in amplifying in the circle with lower left quarter among Fig. 1 like that, described buffer solution flows along the outward flange near the end of each gel-filled kapillary 1, forms the outer surface low 40 (buffer solution stream) that flow into facing to 6 li in the uncovered kapillary of gel kapillary 1.Surface low 40 (buffer solution stream) lining outside the dna fragmentation that goes out from the lower part wash-out of gel-filled kapillary 1 is moved to.Should be noted that,, uncovered kapillary 6 is not arranged that facing to each gel kapillary 1 be can be not problematic in actual applications as long as form mobilely at irradiation position.
Its interior diameter is that the quartz ampoule of 0.1mm is used as the gel-filled kapillary 1 according to present embodiment.Can use pipe according to specific requirement with scope different interior diameters from 0.05 to 0.3mm.Polyacrylamide (total concentration be 5% and crosslinking rate be 3%) be used as the gel that is filled in the kapillary 1, its production method for example, is introduced in analytical chemistry (Analytic Chem.) 64 phase 1221-1225 pages or leaves (1992) by Y.Baba etc.
When the flow rate as the buffer solution of outer surface low is every container 50nl/ second or does not more have problems for a long time.The shape of DNA band and the relation of fluorescence intensity flow rate is best is optimized according to fluorescence detection unit.Fluorescence detection unit 2 is connected with the external container 7 of filling as the buffer solution of outer surface low, and filling is adjusted with the flow rate of control as the buffer solution of outer surface low as the height of the external container 7 of the buffer solution of outer surface low.Laser beam 4 passes unit 2 from the limit.The end of this fluorescence detection unit and uncovered kapillary 6 join, and flow out the buffer solution as outer surface low at uncovered kapillary 6 places.
Gel-filled kapillary 1 and fiber array folder 9 are placed in the top of fluorescence detection unit 2 together, and the following end of gel-filled kapillary 1 is placed in the top 0.5mm place of laser radiation part.The optical path of the laser beam 4 that the end below gel kapillary 1 begins-promptly distance-the best to irradiation position should be 2mm or shorter.If this distance is long, dna fragmentation may mix with the dna fragmentation that moves in the adjacency channel, similar problem perhaps occurs.
The optical fiber 11 that presss from both sides 10 ray-collecting lens or have ray-collecting lens by fiber array be installed in by on the perpendicular plane in gel-filled kapillary 1 and uncovered kapillary 6 formed planes.Distance from the migrating channels to optical fiber between 11 light entrance end is 2 to 3mm, and is spaced apart 0.35 to 2mm between the adjacent gel filled capillary pipe 1, and each optical fiber 11 can not be exposed to adjacent migrating channels like this.
In the present embodiment, provide four to be one group optical fiber for each migrating channels.Optically be connected on each light receiving element of optical line sensor 13 or face sensor by 12, four optical fiber of optical filter, each light receiving element has different transmission wavelength bands.For the sake of simplicity, Fig. 1 only represents between light receiving element and the optical fiber 11 along the connection of two ends on the direction of the laser beam irradiation of kapillary 1 array that is filled up by gel.
For light receiving element and optical fiber are coupled together, use the CCD that has the optical fiber window, perhaps light receiving element and optical fiber are coupled together with glue by man-to-man ratio at the microscopically adjusting position.Also may prearrange and have the top that anchor clamps have fiber array, so as with glue the top be fixed to line sensor and or face sensor on.
For example, FAM (emission wavelength) by ABI company (Applied Biosystem Inc.) sale with 519nm, JFO (emission wavelength) with 548nm, TAMRA (emission wavelength with 587nm) and ROX (emission wavelength with 605nm) are used as the fluorescence of each family of mark, and Ar
+Laser (wavelength with 488nm) and YAG laser (wavelength with 532nm) are used as energizable laser source 5.Four bandpass filter of the center on the emission wavelength of each fluorescence that reach with about 20nm transmission wavelength bandwidth are used as optical filter 12.
When quantity capillaceous is about 100, by a line sensor being divided into four parts and might measuring fluorescence intensity by the wavelength that separates various fluorescent emission wavelength.When having many kapillaries or from the light of an optical fiber, detecting, use face sensor or use that to provide a plurality of line sensors (using four line sensors herein altogether) of optical detection respectively to various fluorescence be effective by the several light receiving elements that adopt big fibre bundle.
Photodetector such as line sensor or face sensor is controlled by driver 14.The detection signal of photodetector is subjected to the analysis of data processor 15, to discern terminal base kind.In addition, for the dna sequencing fluorescence intensity over time.The output of data processor 15 is fed to output unit 16, as CRT, register or plotting apparatus.
Fig. 2 represents to utilize optical fiber light detects and light receiving part couples together method and several example, thereby detects the fluorescence with fluorescently-labeled dna fragmentation.
Fig. 2 (a) represents an example, therein from a launching site 31 (intersection point of migrating channels and illuminated laser spot, emitting fluorescence at that point) fluorescence is collected after it changes directional light into by the lens 8 that have four optical fiber 11, is equipping the optical filter 32 with different transmission wavelength frequency bands on the side of four optical fiber 11.Optical filter 32 is to form film by the light upstream end that electrically non-conductive material is deposited on optical fiber 11 to constitute.In order effectively the photosensitive unit of optical fiber and line sensor to be coupled together, the exit of optical fiber 11 is done the narrow conical fiber that has a different area with the two ends of matching with photosensitive unit or makes optical fiber 11 and is used as and is connected optical fiber.
Fig. 2 (b) illustrates this situation, collects and pass an optical fiber 11 from the fluorescence of a launching site 31 by lens 8 therein.In Fig. 2 (b), only shown a launching site 31, but in actual conditions, many launching site and and the optical fiber of many correspondences of lens 8 couplings are arranged.
Shown four connect optical fiber 11 and light collecting lens 8 can optically be made an integral body in Fig. 2 (a).Also may adopt lens arra, arrange a plurality of lenticules and constitute straight line in array, the interval between them equals four arrangement interval that connect optical fiber.
In this example, make fluorescence on face sensor (dimension sensor) 13, form image by lens 20 ' by Optical Fiber Transmission.During this time, lens 20 make the light from optical fiber connector become directional light, and with by having the visual separation prism 21 that is used for the wave filter that four wavelength select, and image lens 20 ' is used to fluorescent image is focused on the detecting device.
Should be noted that optical fiber 11 and light collecting lens 8 among Fig. 2 (b) can optically make an integral body.And may use lens arra, in lens arra, arrange a plurality of lenticules and form straight line that the interval between them equates with the arrangement interval of optical fiber.
The system that Fig. 2 (c) expression is such, four optical fiber that face toward launching site 31 therein directly lead to the light receiving element of line sensor.Lens 20 and 20 ' and wave filter 22 be arranged on the centre of each bar optical fiber 11, and link to each other with photosensitive unit (light receiving element) on the optical line sensor by optical fiber 23.
Should be noted that above-mentioned description is relevant with the example that adopts four optical fiber, but the quantity of optical fiber is not limited to four: the optical fiber that can use six, eight or any other quantity according to actual needs.
Second embodiment
First embodiment uses a fluorescence detection unit; But the quantity of fluorescence detection unit is not limited to one.In other words, electrophoresis equipment according to the present invention comprises a plurality of lasing light emitters, many optical fiber, one or more photodetector and two or more fluorescence detection units 2, connect two or more gel-filled capillary arrays 1 at two or more fluorescence detection units place, and the inside of sample fragment migration place form a plurality of outside surface low, as shown in Figure 1.
Fig. 3 represents an example, in this example by separating laser beam or by the two or more smooth unit of arranged in series, detecting the light (signal) of the fragment that goes out from a plurality of gel-filled kapillary wash-outs in fact simultaneously.The same with the situation among Fig. 1 (not shown in Figure 3), gel top capillaceous is immersed in the buffer container that is arranged in the upper position place, and be immersed in the buffer container that is arranged in lower position, and electrophoretic voltage is applied to gel-filled two ends capillaceous corresponding to the bottom of the uncovered kapillary 6 of the gel-filled kapillary 1 of each bar.And fluorescence detection unit 2 is connected with the container that filling is used for the buffer solution of outer surface low 7.
In a collection of quantity of paying capillaceous is 96 (being 12 * 8 in this example), identical with the hole count of the titer plate of adjusting dna sample.But and sample adjusts aut.eq. and is complementary and can is more effective and is in operation that to adopt 24 multiple can be easily, the 24th, and in the titer plate two of every round number times.Like this, we are assumed to be a unit to 96 arrays capillaceous that have that band together with plane formula, and constitute a fluorescence detection unit that is connected with this capillary array; We increase the quantity of this fluorescence detection unit 2 as required then.
Might tandem ground arrange that two or more fluoroscopic examination elements 2 make and perhaps might separate other the fluoroscopic examination element of laser beam irradiation of the window 24 that passes adjacent fluorescent element laser beam and make the parallel beam ground that is produced shine two or more fluorescence detection units.
According to the example shown in Fig. 3, come the laser beam 4 on self-excitation light source rank to be separated into laser beam 4-1 and 4-2, and the direction of passing the laser beam 4-1 of this half mirror is changed by mirror 26 by half mirror 25.In series arrange two or more fluorescence detection units 2 along the optical path of laser beam 4-1 and 4-2, can obviously improve throughput like this.Be connected with photodetector 13 attached to the optical fiber on the fluorescence detection unit 2 11 such as line sensor or face sensor.
Should be noted that the photodetector and the light collection device that are used to detect from the fluorescence of moving to the dna fragmentation in the outer surface low zone interconnect according to the distinct methods described in the embodiment 1.And, self-evident two or more optical sensors 13 or the laser of using.
The 3rd embodiment
First and second embodiment adopt kapillary, also might adopt the groove on flat board constitutes to form migrating channels, to replace with gel-filled kapillary.
Fig. 4 represents an example, this example utilize dividing plate or the groove that forming on the flat board between two glass plates 28, constituting running gel, and utilize them as migrating channels.Migrating channels is isolated from each other with the iris type glass (form groove this moment on the one side of glass plate, and this glass plate combines with another piece glass plate) that comprises plastic plate (all fluorocarbon polymer like an elephant teflon, and other polymkeric substance).Realize irradiation in all migrating channels by the mobile DNA of irradiation on the position of the terminal predetermined distance that leaves glass plate by the dna fragmentation from the end that is clipped in two migrating channels between the glass plate being eluted to outer surface low neutralization.
In other words, be eluted to fluorescently-labeled dna fragmentation in the buffer solution stream (outer surface low) of 30 li of fluorescence detection units, and flow with buffer solution.Laser beam 4 shines the dna fragmentation in the migration simultaneously.Be similar to the situation of the capillary array in embodiment 1 and 2, buffer solution stream (outer surface low) is formed on the place near the capillary gel end, with the diffusion of the dna fragmentation that prevents to go out from the migrating channels wash-out.The plate 28 that is used for fixing running gel vertically is provided with, but but their also along continuous straight runs placements.In Fig. 4, outside the dna fragmentation that comes out from gel is moved to downwards the surface low.The gap of buffer solution between the inwall of the plate 28 (glass plate) that is used for fixing running gel and fluorescence detection unit 30 flow downward (filling not shown in the figures be used for outside the container 7 of buffer solution of surface low).
30 li of the fluorescence detection units of surface low outside containing buffer solution and generating therein, fluorescence is connected with unlimited kapillary 6 from the bottom of measurement unit 30, buffer solution flows out from this position, and on the plate 28 (glass plate) of this stationkeeping running gel is installed in a fixed position on the fluorescence detection unit 30, and leave some distances facing to uncovered kapillary 6.On the gap between one end of the terminal and uncovered kapillary 6 of laser beam 4 irradiation migrations onboard.Should be noted that as long as locating formation in the irradiation place flows, is not can not produce practical problems to open capillaries 6 of the terminal corresponding layout of the migration of each migrating channels.
In addition, self-evident, according to embodiment, the layout of second embodiment can be by adopting realizing synthesizing of a plurality of units of the migrating channels produce between two glass plates and fluorescence detection unit.
In embodiment 1,2 and 3, just a side from fluorescence detection unit receives fluorescence.By improving light collection efficiency as if the reflecting material of mirror is placed on the another side all.
In addition, in embodiment 1,2 and 3, make an explanation with the example of the fluorescence of fluorescently-labeled DNA by detection.Except that these examples, also might adopt chemiluminescence emission (chemiluminescence) to detect the DNA frequency band.Under that situation, reagent is placed in the outer surface low of solution detecting chemiluminescence, this chemiluminescence be by the reagent in the outer surface low and from gel the reaction between the sample compound of the dna fragmentation of wash-out launch.
The present invention by adopt one or several line sensor or face sensor and allow in addition on a large scale in have in the electrophoresis system of irradiated region and carry out optical detection; It is that the override of very high throughput is learned measuring system for the system with many migrating channels provides a kind of characteristics.
Compared with many photomultiplier cells tubular construction, line sensor and face sensor are compacter and more cheap; These characteristics have guaranteed the effective ways of the more cheap system of a kind of configuration.
In addition, it allows to increase as required capillary array, thereby a kind of easy-to-use system is provided.
Claims (20)
1, a kind of electrophoresis equipment that detects DNA or other biological substance, it comprises:
(1) many migrating channels that are used to locate the electrophoresis of DNA or other biological material, their end is arranged and is isolated mutually along straight line;
(2) light sources that produce light, the light of generation are along described straight-line pass, and are crossing with all migrating channels;
(3) optical receiving fiber, the end of optical receiving fiber be arranged in and an end of the light of light source and described migrating channels between the corresponding optical axis that is complementary of intersection point on, and the quantity of optical receiving fiber is equal to or greater than the quantity of migrating channels; And
(4) optical detection devices, its other end with each bar optical fiber is connected, and is used to receive light.
According to a kind of electrophoresis equipment of claim 1, it is characterized in that 2, described migrating channels comprises gel-filled kapillary.
According to a kind of electrophoresis equipment of claim 1, it is characterized in that 3, described migrating channels is isolated formation between two glass plates.
According to a kind of electrophoresis equipment of claim 1, it is characterized in that 4, the optical fiber of described reception light receives the fluorescence that sends from described DNA or other biological substance.
According to a kind of electrophoresis equipment of claim 1, it is characterized in that 5, described optical detection device is an one dimension optical sensor or two-dimensional light sensor.
6, according to a kind of electrophoresis equipment of claim 1, it is characterized in that, described optical detection device is an one dimension optical sensor or two-dimensional light sensor, and the other end of described optical fiber optically is connected with a photosensitive unit of one dimension sensor or dimension sensor at least.
According to a kind of electrophoresis equipment of claim 1, it is characterized in that 7, light collecting lens is placed between the end of optical fiber of light that described light source sends and intersection point between the described migrating channels and described reception light.
According to a kind of electrophoresis equipment of claim 1, it is characterized in that 8, an end of the optical fiber of described reception light forms an optical receiver lens.
9, according to a kind of electrophoresis equipment of claim 1, it is characterized in that, between the described other end of the optical fiber of described reception light and described optical detection device, two or more bandpass filter are set.
10, a kind of electrophoresis equipment that is used to detect DNA or other biological substance, it comprises:
(1) many migrating channels that are used to handle the electrophoresis of DNA or other biological material, their end is arranged and is isolated mutually along straight line;
(2) fluorescence detection units that hold an end of described migrating channels thereon;
(3) light sources that produce light, the light of generation are along described straight-line pass, and are crossing with all migrating channels;
(4) optical receiving fiber, the end of optical receiving fiber be arranged in and an end of the light of light source and migrating channels between the corresponding optical axis that is complementary of intersection point on, and the quantity of optical receiving fiber is equal to or greater than the quantity of migrating channels; And
(5) optical detection devices, its other end with each bar optical fiber is connected, and is used to receive light.
According to a kind of electrophoresis equipment of claim 10, it is characterized in that 11, described migrating channels comprises gel-filled kapillary.
According to a kind of electrophoresis equipment of claim 10, it is characterized in that 12, described migrating channels forms between two glass plates.
According to a kind of electrophoresis equipment of claim 10, it is characterized in that 13, the optical fiber of described reception light receives the fluorescence that sends from described DNA or other biological substance.
According to a kind of electrophoresis equipment of claim 10, it is characterized in that 14, described optical detection device is an one dimension optical sensor or two-dimensional light sensor.
15, according to a kind of electrophoresis equipment of claim 10, it is characterized in that, described optical detection device is an one dimension optical sensor or two-dimensional light sensor, and the other end of described optical fiber optically is connected with a photosensitive unit of one dimension sensor or dimension sensor at least.
According to a kind of electrophoresis equipment of claim 10, it is characterized in that 16, light collecting lens is placed between the end of optical fiber of the intersection point of a described end of described light that is produced by described light source and migrating channels and described reception light.
According to a kind of electrophoresis equipment of claim 10, it is characterized in that 17, an end of the optical fiber of described reception light forms an optical receiver lens.
According to a kind of electrophoresis equipment of claim 10, it is characterized in that 18, an end of the optical fiber of described reception light forms a light and collects wave filter.
19, according to a kind of electrophoresis equipment of claim 10, it is characterized in that, between the described other end of the optical fiber of described reception light and described optical detection device, two or more bandpass filter are set.
20, according to a kind of electrophoresis equipment of claim 10, it has the device of a kind of separation from the light of described light source, and shines two or more described smooth unit.
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| JP6002804A JPH07209251A (en) | 1994-01-14 | 1994-01-14 | Electrophoretic device |
| JP002804/94 | 1994-01-14 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN1109597A CN1109597A (en) | 1995-10-04 |
| CN1318847C true CN1318847C (en) | 2007-05-30 |
Family
ID=11539574
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CNB951011944A Expired - Lifetime CN1318847C (en) | 1994-01-14 | 1995-01-13 | Electrophoresis apparatus |
Country Status (3)
| Country | Link |
|---|---|
| JP (1) | JPH07209251A (en) |
| CN (1) | CN1318847C (en) |
| DE (1) | DE19500638A1 (en) |
Families Citing this family (19)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP3563140B2 (en) * | 1995-01-19 | 2004-09-08 | 株式会社日立製作所 | Capillary array electrophoresis device |
| WO1997021259A1 (en) * | 1995-12-06 | 1997-06-12 | Hitachi Metals, Ltd. | Laser resonator, laser device, device applying laser, and method for oscillation of laser beam |
| DE19948391A1 (en) * | 1999-10-07 | 2001-04-12 | Europ Lab Molekularbiolog | Electrophoresis device for analyzing labeled molecules, especially biological molecules |
| GB0219248D0 (en) * | 2002-08-17 | 2002-09-25 | Univ York | OPtical assembly and method for detection of light transmission |
| JP5063616B2 (en) | 2006-02-03 | 2012-10-31 | インテジェニックス インコーポレイテッド | Microfluidic device |
| JP2008249663A (en) * | 2007-03-30 | 2008-10-16 | Kyushu Institute Of Technology | Fluorescence measurement apparatus |
| KR20120031218A (en) | 2009-06-05 | 2012-03-30 | 인터젠엑스 인크. | Universal sample preparation system and use in an integrated analysis system |
| US8512538B2 (en) * | 2010-05-28 | 2013-08-20 | Integenx Inc. | Capillary electrophoresis device |
| WO2012024658A2 (en) | 2010-08-20 | 2012-02-23 | IntegenX, Inc. | Integrated analysis system |
| EP2742347B1 (en) * | 2011-08-12 | 2020-03-04 | Life Technologies Corporation | Apparatuses, methods, computer program products, and kits for hi-throughput glycan analysis |
| US20150136604A1 (en) | 2011-10-21 | 2015-05-21 | Integenx Inc. | Sample preparation, processing and analysis systems |
| US10865440B2 (en) | 2011-10-21 | 2020-12-15 | IntegenX, Inc. | Sample preparation, processing and analysis systems |
| WO2015073999A1 (en) | 2013-11-18 | 2015-05-21 | Integenx Inc. | Cartridges and instruments for sample analysis |
| US10208332B2 (en) | 2014-05-21 | 2019-02-19 | Integenx Inc. | Fluidic cartridge with valve mechanism |
| EP3209410A4 (en) | 2014-10-22 | 2018-05-02 | IntegenX Inc. | Systems and methods for sample preparation, processing and analysis |
| CN107003279B (en) * | 2014-12-16 | 2020-01-10 | 生物辐射实验室股份有限公司 | Horizontal electrophoresis separation device without sealing and extraction gel method without opening box |
| JP6456983B2 (en) | 2015-02-03 | 2019-01-23 | 株式会社日立ハイテクノロジーズ | Multicolor detector |
| JP6975704B2 (en) * | 2015-02-03 | 2021-12-01 | 株式会社日立ハイテク | Multicolor detector |
| JP6747672B2 (en) * | 2015-03-31 | 2020-08-26 | 日本電気株式会社 | Spectroscopic measuring device and spectroscopic measuring method |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4870004A (en) * | 1984-06-08 | 1989-09-26 | Autoseq, Inc. | Apparatus and method of analyzing nucleic acid molecules |
| US5194915A (en) * | 1990-09-20 | 1993-03-16 | Millipore Corporation | Photometric apparatus and process |
-
1994
- 1994-01-14 JP JP6002804A patent/JPH07209251A/en active Pending
-
1995
- 1995-01-11 DE DE1995100638 patent/DE19500638A1/en not_active Withdrawn
- 1995-01-13 CN CNB951011944A patent/CN1318847C/en not_active Expired - Lifetime
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US4870004A (en) * | 1984-06-08 | 1989-09-26 | Autoseq, Inc. | Apparatus and method of analyzing nucleic acid molecules |
| US5194915A (en) * | 1990-09-20 | 1993-03-16 | Millipore Corporation | Photometric apparatus and process |
Also Published As
| Publication number | Publication date |
|---|---|
| CN1109597A (en) | 1995-10-04 |
| DE19500638A1 (en) | 1995-07-20 |
| JPH07209251A (en) | 1995-08-11 |
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