Source: http://www.google.com/patents/US7195699?ie=ISO-8859-1
Timestamp: 2014-07-24 11:58:13
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Patent US7195699 - Used for analyzing polymers, such as DNA, RNA and proteins, and particularly ... - Google PatentsSearch Images Maps Play YouTube News Gmail Drive More »Sign in<nobr>Advanced Patent Search</nobr>PatentsAn electrophoresis member is produced by laying a plurality of capillaries on an adhesive layer born on a support layer to form a capillary layer, laminating thereon a second support layer, and partially removing the first support layer, the first adhesive layer and the second support layer to partially...http://www.google.com/patents/US7195699?utm_source=gb-gplus-sharePatent US7195699 - Used for analyzing polymers, such as DNA, RNA and proteins, and particularly, used in DNA sequencersAdvanced Patent SearchPublication numberUS7195699 B2Publication typeGrantApplication numberUS 10/628,274Publication dateMar 27, 2007Filing dateJul 29, 2003Priority dateAug 26, 2002Fee statusPaidAlso published asUS7736482, US20040035702, US20070205105Publication number10628274, 628274, US 7195699 B2, US 7195699B2, US-B2-7195699, US7195699 B2, US7195699B2InventorsHiroshi Kawazoe, Tomoyuki Kamata, Atsushi Takahashi, Tsuyoshi Sonehara, Tatemi Ido, Kunio HaradaOriginal AssigneeHitachi Chemical Co., Ltd., Hitachi, Ltd.Export CitationBiBTeX, EndNote, RefManPatent Citations (6), Referenced by (2), Classifications (11), Legal Events (2) External Links: USPTO, USPTO Assignment, EspacenetUsed for analyzing polymers, such as DNA, RNA and proteins, and particularly, used in DNA sequencersUS 7195699 B2Abstract An electrophoresis member is produced by laying a plurality of capillaries on an adhesive layer born on a support layer to form a capillary layer, laminating thereon a second support layer, and partially removing the first support layer, the first adhesive layer and the second support layer to partially expose the capillaries to form a window portion for irradiation and detection and a sample injection portion for injecting a sample.
SUMMARY OF THE INVENTION An object of the present invention is to provide an electrophoresis member that has an accurate, compact capillary array, is easy to handle and has high heat releasability. Another object of the invention is to provide an efficient method of producing the electrophoresis member.
the first adhesive layer is positioned on the first support layer, the capillaries lie on the first adhesive layer to form a capillary layer, and the second support layer is positioned on the capillary layer; the capillary layer has a window portion and a sample injection portion including a terminating end of each capillary, the capillaries, at the sample injection portion, being exposed by partially removing one end portion of the first supporter and, at the window portion, being exposed by partially removing another portion of the first supporter; the capillaries, at each of the window portion and the sample injection portion, are arranged so that axes of the capillaries have a parallel, coplanar relationship; the capillaries have, in the window portion, detection parts defined as intersections of the capillaries and a plane intersecting perpendicularly to the axes of the capillaries; and the capillaries have equal lengths between the detection parts and the terminating ends included in the sample injection portion. In a further aspect of the invention, the capillaries are glass capillaries.
the capillary layer has a window portion and a sample injection portion including a terminating end of each capillary, the capillaries, at each of the window portion and the sample injection portion, being arranged so that axes of the capillaries have a parallel, coplanar relationship; the capillaries have, in the window portion, detection parts defined as intersections of the capillaries and a plane intersecting perpendicularly to the axes of the capillaries; and the capillaries have equal lengths between the detection parts and the terminating ends included in the sample injection portion; (2) a step of laminating a second support layer on the capillary layer; and
BRIEF DESCRIPTION OF THE DRAWING FIG. 1 is a side view illustrating a production process of an embodiment according to the invention.
FIG. 5 is an X�X section of FIG. 3 illustrating an optical system for exciting and detecting fluorescence using the electrophoresis apparatus.
DESCRIPTION OF THE PREFERRED EMBODIMENTS FIG. 1 shows an illustration of the production process and layer structure of the electrophoresis member of the invention, FIG. 2 shows an elevation of an electrophoresis member of an embodiment according to the invention, and FIG. 7 shows an elevation of another embodiment. In FIGS. 2, 3 and 7, to show the array of capillaries, a part or all of the unexposed portions of the capillaries are also shown with full lines.
The specifications of the sample injection portion 7 and the window portion 11 including sizes should be designed to fit the objective apparatus. At the sample injection portion 7 and the window portion 11, the first supporter A (that is, the first support layer 1, the first adhesive layer 2, the second adhesive layer 5 and the second support layer 6) is removed to expose the capillaries 4. The capillaries 4 exposed at the sample injection portion 7 and the window portion 11 should align in parallel at equal intervals. The capillaries should have equal distances between the terminating ends 8 in the sample injection portion 7 and the detection parts 9 defined as intersections of the capillaries 4 and a plane intersecting perpendicularly to the axes of the capillaries 4 in the window portion 11. The tolerance on the intervals and distances may be balanced with the measuring accuracy. Tolerance of �0.5% or less is desirable for highly accurate measurements. The allowances may be larger when high accuracy is not required.
EXAMPLES 1 AND 2 Example 1 FIG. 1 shows the production process employed in this example. CAPTON 300H (75 μm thick, trade name, produced by E. I. Du Pont de Nemours and Company) was used as the first support layer 1, and a silicone adhesive sheet S9009 (100 μm thick, trade name, produced by Dow Corning Asia Co., Ltd.) was used as the first adhesive layer 2 on one surface of the first support layer 1. A laying apparatus 3 (produced by Hitachi Chemical Co., Ltd.) that numerically controls applied load and the movement of an X-Y table was used to lay sixteen glass capillaries 4 (4-1�4-16) coated with polyimide resin (Trade name: TSP050150, produced by Polymicro Technologies Co., Ltd., outside diameter: 150 μm, inside diameter: 50 μm) on the first adhesive layer 2 so that the detection parts 9 were 20 cm distant from the terminating ends 8 of the sample injection portion 7. Load of 100 g was applied on the capillaries being laid. CAPTON 300H (75 μm thick, trade name, produced by E. I. Du Pont de Nemours and Company) was used as the second support layer 6, and a silicone adhesive sheet S9009 (100 μm thick, trade name, produced by Dow Corning Asia Co., Ltd.) was used as the second adhesive layer 5 on one surface of the second support layer 6. The second adhesive layer 5 and second support layer 6 were laminated on the laid capillaries 4 using a vacuum laminator. A laser perforator for perforating printed wiring boards with small-diameter holes were used for outside machining in the way that φ0.2 mm holes were made successively at intervals of 0.1 mm with four shots (pulse width: 5 ms) per hole. To expose the capillaries 4 at the window portion 11 including the detection parts 9 and at the sample injection portion 7 (FIG. 2), the first supporter A was cut without damaging the capillaries 4, and then the first support layer 1, the first adhesive layer 2, the second adhesive layer 5 and the second support layer 6 were peeled off by hands. The opening 11-2 measured 10 mm�20 mm in sizes. At a part 11-1 of the window portion including the detection parts 9, the polyimide coating layer covering the capillaries 4 was removed by a ozone asher.
In the product electrophoresis member, all the distances from the detection parts 9 and the terminating ends 8 in the sample injection portion 7 measured 20 cm�0.5 mm. The glass capillaries, at the window portion 11 including the detection parts 9, were aligned at intervals of 150 μm�10 μm, and at the sample injection portion 7, at intervals of 4.5 mm�0.1 mm. The capillaries were free of damage all over the laid area.
As shown in FIG. 2, the first supporter A has the opening 11-2 including the window portion 11 of the capillaries 4. Holes 10 a and 10 b for setting up are made on both sides of the opening 11-2. In this example, the polyimide coating covering the capillaries 4 for protection was removed only at a part of the window portion 11-1 including the detection parts 9, but may be removed all over the window portion 11. At the detection parts 9 in the window portion 11, exciting light is irradiated to the sample drifting in the capillaries, and the resulting excited fluorescence is detected. In the window portion 11 and sample injection portion 7, the capillaries 4 (4-1�4-16) are arranged in the way their axes have a coplanar parallel relationship. One end portion of the first supporter A (the first support layer 1, the first adhesive layer 2, the second adhesive layer 5 and the second support layer 6) was removed to expose under the new edge the capillaries 4 to form the sample injection portion 7 of 10 mm length. The terminating ends 8 aligning on one line were made by cutting the capillaries 4-1�4-16. The sample in the microtiter plate 39 is injected into the capillaries 4-1�4-16 through the terminating ends 8. The upper edge of the first supporter A has a projection 12 only at the part where the capillaries 4 lie, to use the projection 12 for feeding a separation medium.
FIG. 5 is an X�X section of FIG. 3, illustrating the optical system assembled behind the temperature control plate 14 to excite and detect fluorescence. The light source 34 is a helium neon laser of 594 nm wavelength and 8 mW. The laser ray emitted from the laser is diverted into two by the beam splitter 35 and irradiated to both sides of the detection parts 9 by mirrors 33-1, 33-2, 33-3 and prisms 26-1-26-4. The laser ray transmitted through the beam splitter 35 is reflected by the prism 26-1 arranged in the opening 11-2 and irradiated to the detection parts 9 perpendicularly to the axes of the capillaries 4 from a direction parallel to the plane on which the axes are arranged in parallel. The laser ray reflected by the beam splitter 35 is irradiated through the prism 26-2 to the detection parts 9 of the capillaries from a direction opposed to the former laser ray. The laser rays irradiated to the outermost capillaries 4-1 and 4-16 are, as described in Japanese Patent Application Unexamined Publication No. H09-96623 (1997), irradiated to the adjacent capillaries successively by the lens effects of the capillaries without being made divergent. Irradiating from two opposed directions by diverting a laser ray into two cancels the loss due to reflection, and decreases the dispersion of the intensity of the laser ray to �20% or less. The light L emitted by the sample from the capillaries 4 is made into a parallel beam by a camera lens 36 a, and after the removal of background light other than the fluorescence of the sample by a band-pass filter 37, imaged in 1:1 by the second camera lens 36 b on the photoelectric surface of an image pickup element 38. Because the image pickup element 38 pixel is a CCD of 25 μm in pixel pitch, it can separate and detect the fluorescence from each capillary 4.
Example 2 FIG. 7 shows an electrophoresis member of a second example according to the invention. In this example, the glass capillaries are supported before and behind the detection part 9 by a second supporter 40 (made of a plastic) having finely V-grooves for keeping the parallel capillary alignment. This made the intensity of the light irradiated to the capillaries more uniform to decrease its dispersion within �10%. Further, the structure of the measuring system was simplified by removing the first support layer 1, the first adhesive layer 2, the second adhesive layer 5 and the second support layer 6 at all of the part opposing to the sample injection portion 7 of the capillaries 4 and including the detection parts 9. 41 is a cylindrical ferrule having a penetrating hole. The capillaries are bundled and inserted into the hole, and gaps are filled with an adhesive. The cylindrically bundled ends of the capillary array allow easy sealing, enabling filling the electrophoresis medium in a shorter time by applying higher pressure. FIG. 8 is an enlargement of the second supporter 40, wherein (A) is an elevation, (B) is a side view and (C) is a bottom view. As shown in FIG. 8, the second supporter 40 has sixteen V-grooves, and a square groove is made at the part where the detection parts 9 are placed, not to intercept the irradiation.
Patent CitationsCited PatentFiling datePublication dateApplicantTitleUS6048444 *Nov 26, 1997Apr 11, 2000Hitachi, Ltd.Capillary electrophoresis apparatusUS6063251 *May 30, 1997May 16, 2000Spectrumedix CorporationElectrically insulated capillary arrays for electrophoretic applicationsUS6977034 *Dec 4, 2000Dec 20, 2005Hitachi, Ltd.Capillary array and electrophoresis apparatusJP2000131278A * Title not availableJP2001264293A Title not availableJP2001324475A Title not available* Cited by examinerReferenced byCiting PatentFiling datePublication dateApplicantTitleUS8480970Nov 30, 2005Jul 9, 2013Hitachi Chemical Co., Ltd.Analytical pretreatment deviceUS8480971Apr 15, 2011Jul 9, 2013Hitachi Chemical Co., Ltd.Analytical pretreatment deviceClassifications U.S. Classification204/603, 264/154, 204/452, 204/453, 264/139, 204/604International ClassificationG01N21/64, G01N27/447, G01N27/453Cooperative ClassificationG01N27/44782European ClassificationG01N27/447C5Legal EventsDateCodeEventDescriptionAug 26, 2010FPAYFee paymentYear of fee payment: 4Jul 29, 2003ASAssignmentOwner name: HITACHI CHEMICAL CO., LTD., JAPANOwner name: HITACHI, LTD., JAPANFree format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:KAWAZOE, HIROSHI;KAMATA, TOMOYUKI;TAKAHASHI, ATSUSHI;ANDOTHERS;REEL/FRAME:014336/0146Effective date: 20030620RotateOriginal ImageGoogle Home - Sitemap - USPTO Bulk Downloads - Privacy Policy - Terms of Service - About Google Patents - Send FeedbackData provided by IFI CLAIMS Patent Services©2012 Google