US3445360A

US3445360A - Electrophoresis migration chamber and method of using same

Info

Publication number: US3445360A
Application number: US402741A
Authority: US
Inventors: Hugh D Via Jr
Original assignee: HUGH D VIA JR
Current assignee: HUGH D VIA JR
Priority date: 1964-10-09
Filing date: 1964-10-09
Publication date: 1969-05-20
Anticipated expiration: 1986-05-20

Description

y 0, 1969 H. D. VIA, JR 3,445,360

ELECTROPHORESIS MIGRATION CHAMBER AND METHOD OF USING SAME Filed Oct. 9, 1964 Sheet of 2 INVENTOR.

HUGH D. VIA, JR.

May 20, 1969 H. D. VIA, JR

ELECTROPHORESIS MIGRATION CHAMBER AND METHOD OF USING SAME Filed Oct. 9, 1964 FIG. 2

LIQUID LEVEL INVENTOR.

HUGH D. VIA, JR

United States Patent US. Cl. 204-180 4 Claims ABSTRACT OF THE DISCLOSURE An electrolytic apparatus for applying electric current to specimens in open ended tubes in which an open top receptacle is adapted to receive a second open top receptacle thereabove and specimen tubes are mounted in sealing openings in the bottom of the top receptacle with the openings being of substantial vertical axial length with sealing means at each end of the opening so that a specimen tube may be replaced without losing the electrolyte in the top receptacle and the distance between the bottom of the top receptacle and the bottom of the bottom receptacle is such that a specimen tube can be pushed down in the opening but will remain in sealed contact with the bottom of the opening so that a new specimen tube or a plug can be inserted in the upper part of the said opening. The top receptacle may then be removed and the downwardly extending specimen tube removed and thereafter the upwardly extending specimen tube may be pushed down in sealing engagement with both ends of the opening and into contact with the electrolyte in the bottom receptacle.

The present invention relates to electrolytic apparatus and more particularly to electrophoresis apparatus where a proteinaceous material is electrolytically treated to obtain separation of parts thereof and separation of stains applied therein.

Heretofore various types of electrophoresis equipment have been provided but none has been entirely satisfactory in that the manipulation of the equipment was difiicult and the operator frequently would come in contact with the electrolyte with consequent danger from the caustic or corrosive effect of the electrolyte and from electrical currents and also danger of contamination of the electrolyte and of the materials and equipment used in the electrophoresis process.

An object of the present invention is to provide equipment for electrolytic work which avoids the difficulties and inconveniencies of the prior art.

Another object of the present invention is to provide electrophoresis equipment which may use diiferent lengths of specimens and destaining tubes in the disc electrophoresis method.

Another object is to provide positive electrical conductivity to a sample without conductors being placed over the electrolyte.

A further object is to provide electrophoresis equipment which may be used by relatively unskilled persons with assured satisfactory final results.

Another object is to provide a method and equipment for electrolysis whereby a minimum of contamination and danger can occur.

Other and further objects will be apparent as the description proceeds and upon reference to the accompanying drawings wherein:

FIG. 1 is a full scale vertical section taken on broken line 11 of FIG. 2 but including the cover of the electrophoresis equipment showing a first open top cylindrical receptacle resting upon a supporting surface and a second open top cylindrical receptacle resting on the first receptacle with aperture means in the bottom of the second receptacle supporting specimen or destaining tubes extending into both receptacles and electrical connection means for cooperation with electrolytes in the bottom portions of the receptacles for causing current to flow through the conductive material in the specimen or destaining tubes.

FIG. 2 is a plan view of the electrophoresis equipment of FIG. 1 with the cover removed showing the inwardly opening rabbet at the top of the second receptacle.

FIG. 3 is a fragmentary vertical section taken on a broken line similar to line 1-1 of FIG. 2 of the equipment with a spacing ring between the lower and upper receptacles for accommodating longer lengths of destaining tubes.

FIG. 4 is an enlarged fragmentary vertical section of one of the electrodes for conducting current to or from an electrolyte.

FIG. 5 is an enlarged vertical diametrical section through one of the specimen or destaining tube supporting bushings showing a tube being replaced by a plug pushed downwardly with the plug closing the opening in the bushing before the tube is released avoiding loss of electrolyte.

Briefly the present invention comprises a first open top cylindrical receptacle having an inwardly extending rabbet on its upper edge and a second open top receptacle having an outwardly extending rabbet on its bottom edge and an inwardly extending rabbet on its upper edge whereby the second receptacle can be mounted on the first receptacle and a cover closes the top of the second receptacle.

Each receptacle is provided with a centrally positioned electrode extending upwardly from the bottom and elec trically connected to an electrical connection prong on the outer periphery which is adapted to be connected to a source of electric current. The bottom of the second receptacle is provided with tube and plug receiving openings with sealing O-rings whereby a tube having a conductive material therein will carry current from an electrolyte in the first receptacle to an electrolyte in the second receptacle whereby an electrolytic action may take place on the contents of the tube. The tubes may be replaced when the electrolytic action is completed by forcing the tube downwardly with a similar tube or with a plug which similar tube or plug serves to close the opening preventing loss of electrolyte.

The invention is particularly useful in electrophoresis used in determining the characteristics of proteinaceous material for biological research.

Referring more specifically to the drawings, a first open top receptacle 10 includes a bottom 11 supported on three equally spaced supporting feet 12 and a cylindrical wall 13 having an inwardly opening rabbet at its bottom edge cooperating with an upwardly and outwardly opening rabbet in the bottom 11 receiving the lower rabbet of the cylindrical wall with the outer peripheral flange 15 of the rabbet in the bottom projecting beyond the periphery of the cylindrical wall 13, the joint between the bottom 11 and cylindrical Wall 13 is sealed with a suitable adhesive or solvent for the transparent plastic material of which the receptacles are made.

A first center post 16 is mounted in a central aperture in the bottom 11 and carries a 0.013 inch platinum wire 17 having one end of the wire extending into a radial aperture 18 adjacent the top of the post 16 and spiralled around the post 16 for five convolutions passing through an aperture 19 into a hollow cylindrical base 20 of the post 16 and extending downwardly therefrom to beneath the bottom 11 and having its other end electrically connected by a wrapped around and soldered connection 20A to an insulated copper conductor 21 which extends along the bottom surface of bottom.

A seal of plastic material such as epoxy resin B seals the hollow boss 20 and the remaining portion of the aperture through the base 11 and completely encloses the connection 20A.

The conductor 21 passes through an upwardly and outwardly extending aperture 22 in the bottom 11 and the stud projection of the foot 12 extending outwardly of the outer periphery of the cylindrical wall 13 and a resilient banana-type prong 23 is mounted on a wall 24 extending transversely to the axis of a protecting and mounting boss 25 secured to the cylindrical wall in fixed relation for receiving a hollow cooperating tubular connector from a DC source of electric power. A solution of an electrolyte is placed in the bottom of the first receptacle to the liquid level shown and provides a liquid electrode.

As shown in FIG. 1 a second receptacle 26 includes a disc 27 of substantial thickness providing the bottom for the receptacle with said disc being provided with upwardly and downwardly opening rabbets on the periphery forming an intermediate flange 2S therebetween. The upper rabbet receives the lower inwardly opening rabbet of a second cylindrical wall 29 of the upper receptacle being sealed thereto with suitable sealing means. A cover 30 is provided with a downwardly opening rabbet 31 cooperating with the upper inwardly opening rabbet of the cylindrical wall 29 and has a centrally positioned lifting handle or knob 32.

The disc 27 is provided with a central aperture which receives a center post 16A corresponding to center post 16 and has the same electrical connections identified with the postscript A and otherwise corresponding to the electrical connections in the first receptacle thereby providing for a source of opposite polarity electrical energy to a solution of an electrolyte within the second receptacle.

Disc 27 is provided with a plurality of equally spaced apertures with each aperture an equal distance from the center post 16A, each of which apertures receives a flanged bushing 33 in fluid-tight relation with an axial bore and with each flanged bushing being provided with

circumferential grooves

34, 35 on its inner periphery with one groove adjacent the flange at the top of the bushing and the other groove adjacent the bottom of the bushing and each groove receives a sealing O-ring 36 which provides an eifective seal between the periphery of a nonrounded and cylindrical glass tube 37, a rounded end glass tube 38, or a rounded end cylindrical glass plug 39, whereby a liquid seal is maintained between the tubes or plugs and the bushings 33.

The

tubes

37 and 38 are normally filled with a proteinaceous material in a plastic gel or the like which completely fills the tube preventing liquid from passing therethrough. Such proteinaceous material is electrically conductive so that a solution of an electrolyte in the bottom receptacle 10 connected to a source of electrical energy provides electrical contact with the lower end of the contents of the tube while the solution of an eletcrolyte in the top receptacle 26 connected to an opposite polarity source of current provides electrical contact with the contents of the tube with the circuits being complete with the

electrodes

17 and 17A.

In FIG. 3 a cylindrical spacer ring 40 provided with an outwardly opening rabbet at its lower edge and an inwardly opening rabbet at its upper edge and is mounted on the bottom receptacle 10 and carries the top receptacle 26 thereabove, thereby increasing the spacing of the bottom disc 27 of the top receptacle from the bottom 11 of the bottom receptacle so that a greater length specimen or destaining tubes 41 may be used to provide for an increased length of specimen to be electrolytically treated and by the use of additional spacer rings 40, the height can be varied to accommodate any length of tube desired.

In FIG. 1, the tube 37 shown in operative position above the disc 27 a suitable distance and also the bottom end of the tube 37 is above the bottom 11 by a similar distance and this distance is such that when the tube 37 is to be removed, a second tube or a plug 39 may be substituted for the tube 37 in a manner shown in FIG. 5 whereby the tube 37 may be pushed into contact with the bottom 11 and still retain its seal with the O-ring 36 in the bottom groove 34 while the plug 39 or a diiferent filled specimen tube seals the opening by engagement with the O-ring in the top groove 35. Thereafter the upper receptacle 26 is raised and the tube 37 removed by pulling downwardly and placed in a storage location and the plug 39 is pushed downwardly until it engages both O-rings, giving the double seal shown in FIGS. 1 and 3. The change from one tube to another or from one tube to a plug or from a plug to a tube is readily accomplished with practically no loss of the solution of electrolyte and with minimum contact of the operator with the electrolyte since the plug 39 is of sufficient length to have its upper end extend above the liquid level so the operators hands do not get into the electrolyte solution and the O-rings serve to wipe or squeegee off any liquid.

The

cylindrical walls

13 and 29 with cover 30 and ring 40 prevent unintentional contact with the electrolyte solution until the receptacles are separated a sufiicient distance to raise the specimen tubes from the electrolyte solution in the lower receptacle which separation breaks the circuit. Also the length of the tubes with respect to the spacing between bottom 11 and disc 27 prevents accidental removal of a specimen tube or plug as the bottom 11 acts as a stop while the upper end of the tube is still in engagement with the bottom O-ring in groove 34 so that it is practically impossible to unintentionally remove a tube.

It will be apparent that one unit including the upper and lower receptacles can be used for the electrolytic action on the specimen and a second unit can be used to obtain the destaining action thereby increasing the efficiency of the equipment. Also, an extra bottom receptacle or the equivalent may be used to support the top receptacle. The bushings 33 extend below the bottom 27 of the top receptacle and are adapted to support the top receptacle on a flat surface such as a table top serving as feet to support the top receptacle while the top receptacle can be loaded with specimen tubes with the major portion of the tube extending above the bushings 33 and the bottom of the tube lying within the bushing adjacent the bottom thereof. After the top recepacle is placed over the bottom receptacle with or without an intermediate spacing ring 40, the specimen tubes can be pushed down to the proper level by any suitable means such as a plug or the like to avoid contamination of the electrolyte and to avoid contact of the electrolyte with the technicians fingers.

The dimensions are arranged so it is impossible to push out a tube from the bottom when the top receptacle is mounted on the bottom receptacle as the bottom of the bottom receptacle serves as a stop.

It will thus be seen that applicant has provided a new and useful apparatus and method of electrolytically treating material such as proteinaceous specimens in tubes to provide for efiicient operation and ready replacement of one specimen by another.

It will be apparent that changes may be made within the spirit of this invention as defined by the valid scope of the appended claims.

What is claimed is:

1. Electrophoresis equipment comprising a first open top fluid tight cylindrical receptacle, three supporting feet thereunder, said receptacle being provided with an inwardly opening rabbet along its upper edge, a first center post projecting upwardly from said base, a first platinum wire arranged in a helical pattern around said post and extending through said post to below the base of said first receptacle, a first conductor extending from said first platinum electrode to the periphery of said first receptacle, a

banana prong electrode mounted exteriorly of said receptacle and connected to said conductor, a cylindrical spacer ring having an external rabbet at its bottom edge and an internal rabbet at its upper edge for selective mounting on said first receptacle for extending the effective height of said first receptacle, a cylindrical disc having a diameter to be received in the internal rabbet at the top of said first receptacle and to be received in the internal rabbet on the top of said ring, a flange projecting from said cylindrical disc beyond the external diameter of said ring and said first receptacle whereby said disc may be readily applied to and removed from the upper edge of said first receptacle or said ring, a second center post mounted in said disc and projecting upwardly therefrom, a second platinum conductor spiralled around said second post and extending below said disc, a second conductor connected to said second platinum electrode and extending radially outward therefrom, a cylindrical wall having an internal rabbet mounted in fluid tight relation to the periphery and top of said disc and said flange providing with said disc a second receptacle, an electrical prong connection mounted on said cylindrical wall and connected to said second radially extending conductor, a plurality of flanged bushings extending through said disc with their flanges against the upper surface of said disc, said flanges being equally spaced from each other and spaced equal distances from said second post, each bushing being pro vided with a circumferential groove on its inner periphery adjacent each end thereof, and O-ring mounted in each groove, a plurality of tubes and/or plugs mounted in said bushings in fluid tight relation by said O-rings whereby when the tubes are filled with a proteinaceous material, and an electric current is passed through an electrolyte in said first receptacle from said first platinum wire through the material in a tube and through the electrolyte in said second receptacle through said second platinum wire in said second receptacle, an electrolytic action is obtained on the contents in said tubes.

2. Electrophoresis equipment comprising a first open top receptacle, an electrode in said receptacle, a conductor extending from said electrode to the exterior of said receptacle, a spacer having means on its bottom edge for selective mounting on said first receptacle for extending the effective height of said receptacle, a second open top receptacle including a bottom which may be readily applied to and removed from the upper edge of said spacer, a second electrode in said second receptacle, a conductor connected to said second electrode and extending outwardly therefrom, the bottom of said second receptacle having rod and tube receiving openings therein spaced from said second electrode, a plurality of tubes and/or plugs removably mounted in said openings in fluid-tight relation whereby when the tubes are filled with a conductive material, and electric current is passed through conductors and electrolytes in said receptacles, an electrolytic action is obtained on the contents in said tubes.

3. Electrophoresis equipment comprising a first fluidtight receptacle, an electrode in said first receptacle, a first conductor extending from said electrode to the periphery of said first receptacle, a second receptacle mounted on said first receptacle, a second electrode in said second receptacle, a second conductor extending from said second electrode to the periphery of said second receptacle, the bottom of said second receptacle being provided with specimen tube receiving openings, each opening including a circumference groove adjacent an end thereof, a sealing means in said groove, said openings receiving specimen tubes and/or plugs in fluid-tight relation whereby when a conductive liquid is placed in said receptacles to cooperate with the specimens and the tubes are filled with specimens and electric energy applied to said conductors, an electrolytic action is obtained on the contents in said tubes.

4. In a method of using electrophoresis equipment which includes a first fluid-tight open top receptacle, first electrode means in said first receptacle, a. first conductor extending from said first electrode means to the exterior of said first receptacle, a second fluid-tight open top receptacle mounted on said first receptacle, second electrode means in said second receptacle, a second conductor extending from said second electrode means to the exterior of said second receptacle, the bottom of said second receptacle being provided with specimen tube receiving openings of substantial axial length and including sealing means adjacent the top and bottom of each opening for receiving filled specimen tubes and/ or plugs in fluid-tight relation whereby when conductive liquid electrolyte is placed in said receptacles and contacts the filled specimen tubes and electric energy applied to said conductors, an electrolytic action is obtained on the specimens in said tubes, a fluid-tight separation between said receptacles being maintained when replacing the tubes or plugs with other tubes or plugs while preventing passage of the electrolyte from the second receptacle to the first receptacle, said method comprising pushing the top end of the tube or plug being replaced with a replacement tube or plug until the bottom of the replacement tube or plug extends into its specimen tube receiving opening and the tube or plug being replaced strikes the bottom of the first receptacle while the top of said tube being replaced remains in sealed engagement with the bottom sealing means of said opening whereby said opening is maintained closed, removing the said second receptacle from the first receptacle and completely removing the tube or plug being replaced while the replacement tube or plug maintains the fluid-tight sealing with the top sealing means of said opening, moving the replacement tube or plug downwardly into the opening until such tube or plug fully engages both sealing means and then returning the second receptacle to operating position on said first receptacle.

References Cited UNITED STATES PATENTS 3,208,929 9/1965 Raymond et al 204-299 3,290,240 12/ 1966 Neren 204-299 JOHN H. MACK, Primary Examiner.

E. ZAGARELLA, JR., Assistant Examiner.

US. Cl. X.R. 204299