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US 4,375,401 AGranted
Electrophoresis system
OwnerNicholas Catsimpoolasinventor
Life of this patent
Estimating the term…
Abstract
An improved electrophoresis system for simple, rapid and reproducible separations of biological materials such as proteins, nucleic acids, cells, bacteria, cell organelles, etc. is described. The system consists of several subsystems including a first dimensional system having a multi-column rotatable carousel with automatic density gradient maker and sample deposition, a unique fraction collector cassette which can serve as a sample applicator for the second dimension electrophoresis and a slab-gel second-dimensional electrophoresis system. The total system operation is integrated by microprocessor control. Flexibility in the system design and operation allows for running in an analytical or preparative mode at the discretion of the system operator.
Claims18
An integrated electrophoresis system which comprises in combination
- A. a first dimensional electrophoresis sub-system comprising in combination
- (i) carousel means containing a multiplicity of electrophoresis columns rotatable circularly through a multiplicity of stations wherein one or more of said stations is associated with functions consisting of electrophoresis medium deposition, sample deposition, electrophoresis, electrophoresis medium evacuation, and column washing and drying;
- (ii) electrophoresis medium deposition means, said means being in operative fluid flow relationship with each of said electrophoresis column which is positioned in said electrophoresis medium deposition station whereby electrophoresis medium is provided to each such column;
- (iii) sample deposition means, said means being in operative fluid flow relationship with each of said electrophoresis columns which is positioned in said sample deposition station whereby individual samples are provided to the top of the electrophoresis medium in each said column;
- (iv) electrophoresis means comprising power supply means, electrode means and electrical connector means whereby said electrode means are in ionic contact with said electrophoresis medium in each column and said electrical connector means provides continuous electrical connection between said power supply means, and said electrode means while said column is passing between said electrophoresis stations so as to provide a voltage potential across said electrophoresis medium suitable for carrying out electrophoretic separations;
- (v) electrophoresis medium monitoring means, said means being in operative fluid flow relationship with said electrophoresis medium evacuation station whereby the electrophoresis medium after completion of the electrophoresis is removed from said carousel and is monitored for location of desired sample by measurement of one or more selected parameters;
- (vi) column washing and drying means, said means being in fluid flow relationship with said column washing and drying station whereby each column in such station after evacuation of electrophoresis medium is cleaned and prepared for electrophoresis medium deposition at the initial station of the next cycle;
- (vii) fraction collection means, said means being in operative fluid flow relationship with said electrophoresis medium monitoring means whereby said monitored electrophoresis medium is divided into multiple selected discrete fractions and said fraction collection means containing a sample holder means so arranged and constructed as to serve as a sample holder and applicator means for the second dimensional electrophoresis;
- and
- (viii) cooling fluid means, said means being in operative fluid flow relationship with cooling columns which surround each of said electrophoresis columns in said carousel whereby cooling fluid is provided to said cooling columns throughout the entire rotational cycle of said carousel to thereby remove the heat generated by said electrophoresis;
- B. a second dimensional electrophoresis sub-system comprising in combination;
- (i) slab gel holder tank means so arranged and constructed as to be capable of supporting a multiplicity of slab gels suitable for gel electrophoresis, said slab gels being maintained in a position allowing for sample administration employing said sample holder means from the said first dimensional fraction collector means whereby the discrete fractions collected from said first dimensional electrophoresis are applied to said slab gels;
- (ii) power supply means, said means being in electrical contact with said slab gel holder tank means whereby a potential suitable for carrying out gel electrophoresis is established across said gels;
- and
- (iii) cooling fluid means, said means being in operative fluid flow relationship with said slab gel holder tank means whereby heat generated by said gel electrophoresis is removed;
- C. microprocessor control means, said means being in operative electrical contact with the components of each said sub-system so as to provide operative process control over such sub-systems.
13 more elements
A carousel comprising in combination:
- A. a centrally disposed rotatable shaft having an upper end and a lower end, said upper end being attached to an upper collector assembly comprising a lower cylindrical body and an attached upper cylindrical body of smaller circumference than said lower body, said upper body containing a multiplicity of vertical channels passing through said body, said vertical channels being arranged in three concentric rings, each ring containing an equal number of channels and said lower body containing an equivalent number of horizontal channels said horizontal channels being so arranged and constructed as to intersect perpendicularly with a corresponding vertical channel so as to form a multiplicity of fluid pathways from the top of said upper body to the sides of said lower body, and said lower end of said shaft being attached to a lower collector assembly of identical construction as said upper collector assembly but mounted in an inverse manner on said shaft;
- B. a number of double concentric columns attached circumferentially about and outwardly from said shaft, said number of double columns being equal to the number of channels in any one ring in said collector assemblies, each of said double columns consisting of an inner electrophoresis column and an outer coolant column, the upper section of said double columns being in operative attachment with an upper section ring assembly consisting of a lower ring engaging the upper part of said outer column and having a centrally disposed cylindrical cavity of a circumference greater than said inner column and a channel extending from one side of said ring to the bottom of said ring so as to be in fluid flow relationship to the internal portion of said outer column, a middle ring engaging the upper part of said inner column and having a centrally disposed cylindrical chamber, the bottom of said chamber being in open fluid flow contact with the top of the inside of said inner column and a channel extending from one side of said ring to said cylindrical chamber, and an upper ring engaging the top of such middle ring and having a centrally disposed chamber and a channel extending from one side of said ring to said chamber and the lower section of said double columns being in operative attachment with a lower ring assembly consisting of an upper ring engaging the lower part of said outer column and having a centrally disposed cylindrical of a circumference greater than said inner column and a channel extending one side of said ring to the top of said ring so as to be in fluid flow relationship to the internal portion of said outer column, a middle ring engaging the lower part of said inner column and having a centrally disposed cylindrical chamber, the top of said chamber being in open fluid flow contact with the inside bottom of said inner column and the bottom of said chamber being sealed by a cylindrical semi-permeable membrane means which prevents fluid flow but allows passage of ions and a channel extending from one side of said ring to said chamber and a lower ring engaging the bottom of said middle ring and having a centrally disposed chamber, a first channel extending from one side of said ring to the upper portion of such chamber and a second channel extending from said side of said ring to the lower portion of such chamber;
- C. fluid conduit means extending spoke like from each of the channels opening on the sides around each of said upper and lower collector assemblies to the channels in each of the said upper section rings and lower section rings to provide fluid flow connection between such channels;
- D. fluid flow control means engaging each of said upper and lower collector assemblies comprising a non-rotatable port means having a number of vertical channels so arranged and constructed so that at least one of said vertical channels is in a position opposing the channel rings contained in said collector assemblies, stationary disk means associated with said non-rotatable port means said disk having grooves or holes arranged around concentric rings said grooves or holes being of pre-selected patterns and being so arranged and constructed to oppose the channel rings on said port means, rotating disk means associated with said collector assembly and being slidably engaged with said stationary disk means said rotating disk means having a multiplicity of holes arranged in concentric rings, the number of holes in each such ring being equal to the number of inner columns and the number of channels in each ring of said collector assembly, whereby as said collector assembly and associated disk rotate, their holes and directly opposed channels are either in operative fluid flow relation with holes or groves in said stationary disk means and the opposing channels in said port means or said fluid flow is blocked thus providing pre-selected fluid flow patterns in said channels.
1 more element
Dependencies are inferred from the claim text.