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Claims  |
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What is claimed is:
1. Capillary electrophoresis apparatus comprising a capillary tube of the
type which can be electrically charged, said capillary tube having first
and second ends,
first means at said first end of said capillary tube providing a source of
buffer solution and a source of a sample substance to be analyzed,
second means coupled to said apparatus for applying electrical potential
across said capillary tube whereby a sample flows through said capillary
tube and past a detector,
said first means includes a rotatable table carrying a plurality of sample
cups and a holder for holding an end of said capillary tube in operative
relation with one of the said cups, said cups containing either buffer
solution or a sample to be analyzed, and
a T-shaped section of capillary tube inserted in said capillary tube and
coupled to a source of cleaning fluid usable to clean said capillary tube.
2. The apparatus defined in claim 1 and including a removable coiled
capillary cartridge coupled to said capillary tube.
3. The apparatus defined in claim 1 wherein said capillary tube comprises a
plurality of sections coupled together end to end, the internal surfaces
of said sections coated with different chemicals.
4. The apparatus defined in claim 3 wherein said chemicals serve to modify
the flow of sample material along a capillary tube.
5. The apparatus defined in claim 3 wherein the adjacent ends of said
sections are butted together and are secured together by means of
surrounding a sleeve which is secured to outer surfaces of said capillary
sections.
6. The apparatus defined in claim 1 and including a source of an inert gas
coupled to said rotatable table for addition to sample cups therein.
7. The apparatus defined in claim 1 and including a detector coupled to the
output end of said capillary tube for detecting the passage of sample
therethrough.
8. Capillary electrophoresis apparatus comprising a capillary tube of the
type which can be electrically charged, said capillary tube having first
and second ends,
first means at said first end of said capillary tube providing a source of
buffer solution and a source of a sample substance to be analyzed,
second means coupled to said apparatus for applying electrical potential
across said capillary tube whereby a sample flows through said capillary
tube and past said detector,
said first means includes a rotatable table carrying a plurality of sample
cups and a holder for holding an end of said capillary tube in operative
relation with one of the said cups, said cups containing either buffer
solution or a sample to be analyzed,
said table including an annular array of holes in which sample cups are
seated,
said holder being disposed so that said first end of said capillary tube is
always positioned over one of said sample cups,
said holder including a cylindrical portion which has an aperture which is
adapted to receive said first end of said capillary tube and said holder
further has a hollow tubular portion which extends downwardly from said
cylindrical portion and is adapted to be inserted into a sample cup,
said holder having a second aperture to receive an electrode,
third means at said second end of said capillary tube including a rotatable
table and a plurality of second sample cups carried thereby, a second
holder for holding said second end of said capillary tube in operative
relation with one of said second sample cups,
a first electrode in operative relation with said first end of said
capillary tube and a second electrode in operative relation with said
second end of said capillary tube, and means for applying electrical
potential between said first and second electrodes,
said electrical potential being applied with said first and second ends of
said capillary tube seated in buffer solution,
a vertical post disposed adjacent to said holder and having a slot in its
side wall,
a vertical rod inside said vertical post and adapted to move vertically up
and down,
a horizontal arm extending laterally from said vertical rod through said
slot,
a first electrode on the outer wall of said vertical rod,
a lead extending from said first electrode along said horizontal arm into
said second aperture in said holder and into said hollow tubular portion,
and
a second electrode disposed within said vertical post and including a lead
coupled to a power supply,
said first electrode and said second electrode being positioned so that
when said vertical rod moves a predetermined distance, said first and
second electrodes make contact with each other.
9. The apparatus defined in claim 8 wherein first electrode is a film
electrode.
10. The apparatus defined in claim 8 wherein said capillary tube having an
inside diameter in the range of 25 microns to about 200 microns.
11. The apparatus defined in claim 8 and including table drive means for
rotating said table with respect to said holder.
12. The apparatus defined in claim 11 and including electronic circuit for
automatically synchronizing the operation of said holder drive means and
said table drive means so that in a sequence of operations, said holder is
raised above said table and then said table is rotated to a position where
a selected cup is beneath said holder, and then said holder is lowered so
that said first end of said capillary tube is in said cup.
13. Capillary electrophoresis apparatus comprising a capillary tube of the
type which can be electrically charged, said capillary tube having first
and second ends,
first means at said first end of said capillary tube providing a source of
buffer solution and a source of a sample substance to be analyzed,
second means coupled to said apparatus for applying electrical potential
across said capillary tube whereby a sample flows through said capillary
tube and past said detector,
said first means includes a rotatable table carrying a plurality of sample
cups and a holder for holding an end of said capillary tube in operative
relation with one of the said cups, said cups containing either buffer
solution or a sample to be analyzed, and
said second means includes a second rotatable table carrying a plurality of
sample cups and a second holder for holding the other end of said
capillary tube, and
means for varying the spacing between said first and second holders and the
operating length of the capillary tube therebetween.
14. An automated capillary electrophoresis system including
a computer for controlling the operation of the system as an
electrophoresis system,
a bus connected to said a computer,
a capillary tube for receiving a sample of a substance to be analyzed and
having an input end and an output end, PG,47
a detector adjacent to said capillary tube for detecting the passage of
elements of said sample to be analyzed thereby,
a first electorde at said input end of said capillary tube and in operative
relation therewith and with fluids contained therein,
a second electrode at said output end of said capillary tube and in
operative relation therewith and with fluids contained therein,
a power supply having terminals adapted to provide a plurality of output
voltages, said terminals being coupled to said electrodes for applying
voltages therebetween,
said power supply also being connected to said bus and to said computer
whereby its operation and the application of selected voltages to said
electrodes can be controlled and sequenced,
a timer control coupled to said bus and to said computer for controlling
the time duration of the application of operating voltages to said
electrodes by said power supply,
a first holder for said input end of said capillary tube,
a second holder for said output end of said capillary tube,
first holder drive means coupled mechanically to said first holder for
driving said first holder and said input end of said capillary tube
vertically up and down, said first holder drive means being electrically
coupled to said bus and to said computer whereby the operation thereof can
be automatically controlled,
first holder drive sensor means in operative relation with said first
holder drive means for controlling the movement thereof,
second holder drive means coupled mechanically to said second holder for
driving said second holder and said output end of said capillary tube
vertically up and down, said second holder drive being electrically
coupled to said bus and to said computer whereby the operation thereof can
be automatically controlled,
second holder drive sensor means in operative relation with said second
holder drive means controlling the up and down movement thereof,
a first rotatable table adjacent to said first holder and carrying cups
positioned to receive said input end of said capillary tube,
a second rotatable adjacent to said second holder and carrying cups
positioned to receive said output end of said capillary tube,
a first table drive means coupled mechanically to said first rotary table
and electronically to said bus and to said computer,
first table drive sensor means associated with said first table drive means
to control the operation thereof,
a second table drive means coupled mechanically to said second rotary table
and electrically to said bus and to said computer,
second table drive sensor means assoaciated with said second table drive
means to control the operation thereof,
a clock control means in said computer,
said computer operating to:
1. turn on said first and second holder drive means to raise said first and
second holders a predetermined amount which raises said first and second
holders above said first and second rotary tables respectively,
2. turn on said first and second table drive means to rotate said first and
second tables to a starting position under the control of said first and
second table drive sensor means,
3. turn on said first and second holder drive means to lower said first and
second holders an amount determined by said first and second holder drive
sensor means which places said holders and said input end of said
capillary tube, which is filled with buffer solution, in a first cup
containing a sample substance to be analyzed,
4. cause a sample fluid to be injected into said capillary tube; and
5. cause said sample to flow along said capillary tube.
15. Capillary electrophoresis apparatus comprising a box-like support
member having a top wall, front and rear walls and left and right end
walls,
a first box enclosure at one end of said support member,
a second box enclosure at the opposite end of said support member,
said box enclosures each including a base member, left and right sidewalls,
front and rear walls and a removable cover,
a first rotatable table disposed within said first box enclosure and
rotatably supported on the base member thereof,
a plurality of apertures in said first table and a sample cup seated in
each of said apertures,
a vertically moveable first post disposed adjacent to said first table and
including a first hollow tube extending downwardly therefrom over said
apertures in said first table, said first hollow being adapted to be moved
into and out of a sample cup when said first arm moves down and up,
a first electrode extending into said first hollow tube,
a second rotatable table disposed within said second box enclosure and
rotatably supported on the base member thereof,
a plurality of apertures in said second table and a sample cup seated in
each of said apertures,
a vertically moveable second post disposed adjacent to said second table
and including a second hollow tube extending downwardly therefrom over
said apertures in said second table,
said hollow tube being adapted to be moved into and out of a sample cup
when said second arm moves down and up,
a second electrode extending into said second hollow tube,
said first and second electrodes being adapted to have electrical power
coupled thereto,
a capillary tube extending from said first box enclosure to said second box
enclosure and having one end disposed within said first tube and the other
end of said capillary tube disposed within said second hollow tube,
a detector disposed adjacent to said capillary tube for detecting the
passage of charged particles therethrough, and
a pump coupled to the output end of said capillary tube, said pump
comprising a miniature pump mounted inside said housing.
16. The apparatus defined in claim 15 wherein said rotatable table is
supported on a vertical post which extends below the top wall of said
support member and includes a lower post portion thereat,
a motor coupled to said lower post portion beneath said top wall of said
support member,
a horizontal sensor disk secured to said lower post portion and adapted to
be rotated thereby, and
an optical sensor coupled to said horizontal disk and adapted to operate
said motor in response to the position of said sensor disk with respect to
said sensor.
17. The apparatus defined in claim 16 wherein said motor is supported on
the lower surface of said top wall of said support member.
18. The apparatus defined in claim 15 and including power interlock means
associated with the covers of said boxes for disconnecting electrical
power when a cover is removed.
19. The apparatus defined in claim 15 and including a first hollow, tubular
vertical post having a slot in its wall with said vertically moveable
first post disposed therewithin,
a first horizontal arm extending from said vertically moveable first post
through said slot and carrying said first hollow tube at its outer end
said vertically moveable first post having a lower post portion extending
downwardly through the base member of said first box to beneath said top
wall of said support member,
a first motor coupled to said lower post position to drive it up and down,
a first optical sensor positioned adjacent to said lower post position,
a first means carried on said lower post position and positioned to operate
with said first optical sensor to contorl the operation of said first
motor,
a second hollow tubular vertical post having a slot in its wall with said
vertically moveable second post disposed therewithin,
a second horizontal arm extending from said vertically moveable second post
carrying said second hollow tube at its outer end, said vertically
moveable second post having a lower portion extending downwardly through
the base member of said second box to beneath said top wall of said
support member,
a second motor coupled to said lower post portion of said vertically
moveable second arm to drive it up and down,
a second optical sensor positioned adjacent to said lower post portion of
said vertically moveable second arm, and
a second means carried on said lower post portion of said vertically
moveable second arm and positioned to operate with said second optical
sensor to control the operation of said second motor.
20. Capillary electrophoresis apparatus comprising a box-like support
member having a top wall, front and rear walls and left and right end
walls,
said top wall having a top surface and a bottom surface,
a first box enclosure at one end of said support member,
a second box enclosure at the opposite end of said support member,
said box enclosures each including a base member, left and right sidewalls,
front and rear walls and a removable cover,
an aperture in said top wall of said box-like support member, said aperture
extending along a portion of the length at said top wall,
a first rotatable table disposed within said first box enclosure and
rotatably supported on the base member thereof,
a plurality of apertures in said first table and a sample cup seated in
each of said apertures,
a second rotatable table disposed within said second box enclosure and
rotatably supported on the base member thereof,
a plurality of apertures in said second table and a sample cup seated in
each of said apertures,
a capillary tube extending from said first box enclosure to said second box
enclosure and having one end disposed in operative relation with said
first table and the other end of said capillary tube disposed in operative
relation with said second table, and
a detector disposed adjacent to said capillary tube for detecting the
passage of charged particles therethrough,
said first box being mounted on said top wall,
said second box being slidably mounted on said top wall and secured to
mounting means disposed beneath said top wall and within said box-like
support member.
21. The apparatus defined in claim 20 wherein said last-named means
includes
a first rotatable wheel and a second rotatable wheel spaced from said first
rotatable wheel,
a drive belt coupled to said rotatable wheels,
a drive motor coupled to said first rotatable wheel, and
means coupled to said belt and said second box whereby operation of said
drive motor and movement of said drive belt cause said second box to slide
along said top wall to assure different spacings from said first box.
22. The apparatus defined in claim 21 and including a bellowtype screen
disposed along said aperture in said top wall to prevent foreign objects
from falling into said box-like support member.
23. Capillary electrophoresis apparatus comprising a box-like support
member having a top wall, front and rear walls, and left and right end
walls,
said top wall having a top surface and a bottom surface,
a first box enclosure at one end of said top wall of said support member,
a second box enclosure at the opposite end of said top wall of said support
member,
means supporting a capillary tube between said first box and said second
box, and
means associated with said box-like support member for varying the spacing
between said first box and said second box whereby capillary tubes of
different lengths can be supported therebetween.
24. Capillary electrophoresis apparatus comprising a capillary tube of the
type which can be electrically charged, said capillary tube having first
and second ends,
first means at said first end of said capillary tube providing a source of
buffer solution and a source of a sample substance to be analyzed,
second means coupled to said apparatus for applying electrical potential
across said capillary tube whereby a sample flows through said capillary
tube and past said detector,
said first means includes a rotatable table carrying a plurality of sample
cups and a holder for holding an end of said capillary tube in operative
relation with one of the said cups, said cups containing either buffer
solutions or a sample to be analyzed,
a magnifying glass coupled to said capillary tube for viewing the flow of
fluid through said capillary tube, and
a pump coupled to the output end of said capillary tube.
25. The apparatus defined in claim 24 wherein said magnifying glass permits
observation of the sample meniscus in said capillary tube to determine
electroosmotic flow therein.
26. The apparatus defined in claim 24 wherein said magnifying glass permits
observation of the sample meniscus in said capillary tube to determine
osmotic flow therein.
27. Capillary electrophoresis apparatus comprising a capillary tube of the
type which can be electrically charged, said capillary tube having first
and second ends,
first means at said first end of said capillary tube providing a source of
buffer solution and a source of a sample substance to be analyzed,
second means coupled to said apparatus for applying electrical potential
across said capillary tube whereby a sample flows through said capillary
tube and past said detector,
said first means includes a rotatable table carrying a plurality of sample
cups and a holder for holding an end of said capillary tube in operative
relation with one of the said cups, said cups containing either buffer
solution or a sample to be analyzed, and
a source of an oxygen absorbent gas coupled to said rotatable table for
introduction of said gas into sample cups to remove air bubbles therefrom.
28. The apparatus defined in claim 27 wherein said gas is a gas selected
from the group of gases including helium, nitrogen and argon.
29. Capillary electrophoresis apparatus comprising a capillary tube of the
type which can be electrically charged, said capillary tube having first
and second ends,
first means at said first end of said capillary tube providing a source of
buffer solution and a source of a sample substance to be analyzed,
second means coupled to said apparatus for applying electrical potential
across said capillary tube whereby a sample flows through said capillary
tube and past said detector,
said first means includes a rotatable table carrying a plurality of sample
cups and a holder for holding an end of said capillary tube in operative
relation with one of the said cups, said cups containing either buffer
solution or a sample to be analyzed, and
including means for operating said second means to cause said electrical
potential to be cyclically reversed to cause said sample to move first in
one direction and then in the opposite direction past said detector to
provide a series of detector pulses for identification of the sample.
30. The method of detecting the nature of a sample of a substance by
capillary electrophoresis comprising the steps of
passing a substance to be analyzed in one direction through a detector to
obtain a pulse representative of the optical wavelength of said sample,
passing said substance in the opposite direction through said detector to
obtain a second pulse representat0ve of the optical wavelength of said
sample and,
continuing the process of passing said substance back and forth through
said detector to obtain a series of pulses which combine to provide
accurate identification of the sample.
31. Apparatus for analyzing a sample material comprising
a capillary tube
a detector adjacent to said capillary tube and adapted to provide an output
detector pulse when a sample material passes along said capillary tube and
through said detector,
a source of sample material at one end of said capillary tube to be passed
down said tube to said detector,
electrical means connected across said capillary tube for causing said
sample material to pass in a first direction along said capillary tube
through said detector to provide a first detection pulse,
said electrical means being reversible to cause said sample material to
flow in the reverse direction to said first direction to cause said sample
material to pass through said detector to provide a second detector pulse,
and
means for toggling said electrical means to cause said sample substance to
pass back and forth through said detector and providing thereby multiple
detector pulses.
32. Capillary electrophoresis apparatus comprising a capillary tube of the
type which can be electrically charged, said capillary tube having first
and second ends,
first means at said first end of said capillary tube providing a source of
buffer solution and a source of a sample substance to be analyzed,
second means coupled to said apparatus for applying electrical potential
across said capillary tube whereby a sample flows through said capillary
tube and past said detector,
said first means includes a rotatable table carrying a plurality of sample
cups and a holder for holding an end of said capillary tube in operative
relation with one of the said cups, said cups containing either buffer
solution or a sample to be analyzed, and
said capillary tube is in the form of a coil of glass tubing.
33. Capillary electrophoresis apparatus including a capillary tube having
an inlet end and an outlet end for receiving a sample to be identified and
for passing the sample along the length of the capillary,
a porous glass tube receiving the outlet end of said capillary tube and
an auxiliary tube in said porous glass tube having one end adjacent to the
outlet end of said capillary tube and adapted to receive a sample from
said capillary tube, said auxiliary tube having an outlet end from which
the sample exits to a collection means.
34. The apparatus defined in claim 33 wherein the outlet end of said
auxiliary tube is positioned over a rotatable table, for receiving samples
from said auxiliary tube.
35. The apparatus defined in claim 34 and including a sensor at the outlet
end of said auxiliary tube, an optical glass fiber coupled at one end to
said sensor and at the other end to a detector for detecting the passage
of a sample through said auxiliary tube whereby rotation of said rotatable
table may be effected as each sample appear.
36. Capillary electrophoresis apparatus comprising
a capillary tube of the type which can be electrically charged, said
capillary tube having first and second ends,
first means at said first end of said capillary tube providing a source of
buffer solution and a source of a sample substance to be analyzed,
second means coupled to said apparatus for applying electrical potential
across said capillary tube whereby a sample flows through said capillary
tube and past a detector,
said first means including a rotatable table carrying a plurality of sample
cups and a holder for holding anend of said capillary tube in
operativerelation with one of said cups, said cups containing either
buffer solution or a sample to be analyzed, and
a plurality of chemical substances spaced apart along the length of said
capillary tube, each of said chemical substances having a specific action
with respect to constituent molecules contained within a sample flowing
along said capillary and either attracting a molucule for later elution or
permitting a molecule to pass along the capillary tube.
37. Capillary electrophoresis apparatus comprising
a capillary tube of the type which can be electrically charged, said
capillary tube having first and second ends,
first means at said first end of said capillary tube providing a source of
buffer solution and a source of a sample substance to be analyzed,
second means coupled to said apparatus for applying electrical potential
across said capillary tube whereby a sample flows through said capillary
tube and past a detector, and
a plurality of chemical substances spaced apart along the length of said
capillary tube, each of said chemical substances having a specific action
with respect to constituent molecules contained within a sample flowing
along said capillary and either attracting a molucule for later elution or
permitting a molecule to pass along the capillary tube.
38. The apparatus defined in claim 37 wherein said chemical substances are
in the form of coatings on the wall of said capillary tube.
39. The apparatus defined in claim 37 wherein said chemical substances are
in bulk form and disposed within said capillary tube.
40. The apparatus defined in claim 39 wherein said chemical substances are
coated on globules of insulating material. |
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Claims |
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Description |
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BACKGROUND OF THE INVENTION
Electrophoresis is a phenomenon in which charged particles move in a
conductive buffer medium or fluid across which a potential difference is
applied. The migration is toward an electrode carrying charge opposite to
that of the particles.
Electrophoresis is one of the most important methods available for the
investigation of biological materials, and probably the most efficient
procedure for the separation and detection of proteins and other matter.
Electrophoresis separation relies on the differential speeds of the
migration of differently charged particles in an electrical field. The
migration speed is primary a function of the charge on the particle and
the field strength applied and the charge on a particle is determined by
the pH of the buffer medium. The most important application of this
technique in biomedical research and clinical chemistry laboratories, is
in the electrophoretic separation of proteins, nucleic acids, their
component peptides and oligonucleotides, as well as complex macromolecules
such as lipoproteins.
Several different systems are known for practicing electrophoretic
separation. For example, one system, known as zonal procedures, has
advantages but it also has certain limitations. Some of the most common
limitations are: The amount of sample required in order to reveal the
components by the common staining procedures is usually large, the
preparation of the apparatus and complete system involved in the
electrophoretic separation is commonly tedious and time consuming, the
time required to obtain complete separation of the components is often
hours, the time required to reveal the components and to obtain some
quantitation of the separated substances is also commonly hours, the yield
of recovery of the components as biological actives in most cases is very
low, the reproducibility of the electrophoretic separation is not 100
percent accurate, and the automation to perform the entire system
operation is almost lacking.
Capillary electrophoresis has been shown to be a technique for obtaining
high separation efficiency. For some proteins and small peptides,
separation efficiencies of approximately one million to about a few
million have been demonstrated. In general, this technique utilizes a
fused silica (quartz) capillary with an inside diameter ranging from about
25 microns to about 200 microns, and a length ranging from about 10
centimeters to about 100 centimeters. Since the entire volume of the
column is only 0.5 to about 30 microliters (yielding probably the smallest
total surface area of column chromatography), the injection volume is
usually in the low nanoliters range. As a consequence, the sensitivity of
this technique is quite high and it is possible to obtain quantitation in
the order of picomoles (and probably femtomoles or attomoles) using
fluorescence, electrochemical, laser induced fluorescence, and mass
spectrometry detectors, and to obtain quantitation in the order of
nanomoles using ultraviolet detectors.
In capillary electrophoresis, the efficient heat transfer from small
diameter capillaries permits application of unusually high voltages
ranging from about 5,000 volts to about 30,000 volts while maintaining a
low current, in the range of about 10 microamperes to about 90
microamperes. The application of high voltages promotes more effective
separations and increases the speed of analysis to record times of about 5
to 40 minutes.
In addition to high separation efficiency (theoretical plates), fairly high
resolution, high sensitivity quantitation, and small migration (retention)
times, capillary electrophoresis presents a few more advantages over
conventional electrophoresis, and in general, other chromatographic
procedures. Some of these advantages are: a) application to a wide variety
of samples ranging from small ions to proteins or other macromolecules of
molecular weights of approximately 290,000 daltons or higher (such as DNA
fragments, viruses, and subcellular particles) by using essentially the
same column and probably the same conditions of electrophoretic
separation; b) capillaries should provide an ideal system to explore
nonaqueous media, particularly with substances which are highly
hydrophobic; c) capillaries are reusable many times making the
electrophoretic separation system very practical and economical; d)
on-line electronic detection permits good quantitation and further
enhances possibilities for fully automatic operation making the capillary
electrophoresis system of higher resolution, greater speed, and better
accuracy than conventional methods.
In the prior art, it is generally known that a material, containing
mixtures of substances to be analyzed, can be passed along a capillary
tube and through a detector under the influence of an applied voltage. The
applied voltage charges the substances and the charges on the substances
determine their spacing and their speed of passage along the capillary
tube.
The prior art, U.S. Pat. Nos. 3,620,958, 3,948,753 and 4,459,198, show
electrophoresis apparatus including a capillary tube connected between two
containers for containing the substance to be analyzed and having
electrical potential applied between the two containers and across the
capillary tube. While the various forms of apparatus shown in these
patents are apparently useful, they require large concentrations of
samples to be analyzed and none is capable of being automated or provides
teaching related to automation.
The present invention provides high voltage capillary electrophoresis
apparatus including, among other things, means for feeding small
concentrations of sample material into a capillary tube, automatically
applying the proper voltage to cause the components of the sample to be
charged and to flow along the capillary tube through a detector wherein
the components are detected and a printed record is made. The apparatus
can then automatically repeat the process for the analysis of multiple
samples.
The basic apparatus of the invention is susceptible of many modifications
in its various parts including the capillary tube portion. In addition,
the method of detection of samples may be varied and the collection of
samples can be modified. The invention can also be adapted to measure
electroosmotic flow in a capillary.
DESCRIPTION OF THE DRAWINGS
FIG. 1 is a perspective view of the rear of apparatus embodying the
invention;
FIG. 2 is a sectional view along the lines 2--2 in FIG. 1;
FIG. 3 is a perspective view of a portion of the apparatus shown in FIG. 1;
FIG. 4 is a perspective view of a portion of the apparatus of FIG. 1;
FIG. 5 is a sectional, elevational view of a portion of the apparatus of
FIG. 1;
FIG. 6 is an enlarged perspective view of a portion of the apparatus of
FIG. 1;
FIG. 7 is a front view of a portion of the apparatus of FIG. 1;
FIG. 8 is a side elevational view of a modification of a portion of the
apparatus of FIG. 1 with portions thereof in section;
FIG. 9 is a front elevational view of a modification of a portion of the
apparatus shown in FIG. 1;
FIG. 10 is a schematic representation of the electronic control system used
in the invention;
FIG. 11 is a sectional view of a modification of the capillary portion of
the apparatus of FIG. 1;
FIG. 12 is a plan view of a portion of a detector used in FIG. 1 with the
apparatus of FIG. 11;
FIG. 13 is a schematic representation of a mode of operating multiple
pieces of apparatus of the type shown in FIG. 1;
FIGS. 14A and 14B together are a bottom view of a modification of the base
support of the invention;
FIG. 15 is perspective front view of a modification of the invention;
FIG. 16 is a perspective view of a modified capillary cartridge usable with
the apparatus of the invention;
FIG. 17 is a perspective of apparatus embodying modifications of the
invention;
FIG. 18 is a perspective enlarged view of a switch used with the apparatus
of FIG. 17;
FIG. 19 is a front elevational view of a portion of apparatus of the
invention illustrating a modification thereof;
FIG. 20 is a front elevational view of a modification of the apparatus
shown in FIG. 19;
FIG. 21 is a front elevational view of a portion of a capillary tube and
magnifying glass used to analyze electroosmotic flow;
FIG. 22 is a perspective view of a modification of apparatus embodying the
invention;
FIG. 23 is a modification of the capillary tube used with the invention;
FIG. 24 is a front elevational view illustrating additional apparatus which
can be used in the apparatus of the invention;
FIG. 25 is a perspective view of a modification of the invention used with
some of the apparatus of FIG. 24;
FIG. 26 is a perspective view of a modification of the apparatus of FIG.
25;
FIG. 27 is a front elevational view of a portion of the apparatus of the
invention illustrating a mode of operation thereof;
FIG. 28 shows pulses detected by the method of FIG. 27;
FIG. 29 shows a curve derived from the pulses of FIG. 28; and
FIG. 30 is a side elevational view of a modified capillary including means
for cleaning the capillary.
DESCRIPTION OF THE INVENTION
The automated electrophoresis apparatus of the invention 10, shown from the
rear in FIG. 1, includes a base support member to which various pieces of
operating equipment are secured. The support member 20 is box-like and
includes a top wall 30, a front wall 40, a rear wall 50 and end walls 60
and 70 all of which extend downwardly from the top wall. A bottom cover
plate 80 (FIGS. 1 and 2) is secured to the support member 20 and provides
a flat support surface for the apparatus 10.
The support member 20 is of metal or a plastic and carries on top wall 30 a
left hand box 90 and a right hand box 100 as seen in FIG. 1. The left hand
box 90 includes an insulating base plate 110, of a metal or plastic,
secured to top wall 30 and a transparent enclosure, of plexiglass or the
like, including (FIGS. 1 and 2) left and right side walls 120 and 130,
front and rear walls 140 and 150 and a top wall 160. The top wall 160 is a
cover for the box 90 and is adapted to be lifted off the box by means of
knob 162 to provide access to the interior thereof. The enclosure for box
90 is suitably secured to the base 110.
Box 90 is provided with a rotatable horizontal table 170 having a circular
array of holes or apertures 180 in which fluid sample cups 190 are seated.
The table is detachably secured to the upper end of a vertical post 200 so
that tables with different numbers of holes or different sizes of holes or
with other features can be secured to the post. The post 200 extends
through and beneath the top wall 30 of the support member 20 (FIGS. 2 and
3) where it is suitable connected to a small motor 210 which is used to
rotate the post 200 and table 170. The motor 210 is secured to the lower
surface 32 of the top wall 30 and is of a type which permits the post 200
or an extension thereof to extend through it to be driven thereby.
The lower end of the post 200 carries a horizontal disk 203 (FIG. 3) which
rotates with the post and includes a slot 205 which is adapted to operate
with an optical sensor 212 positioned adjacent thereto.
Adjacent to the rotatable table 170, referring to FIGS. 1, 2 and 6, is a
hollow, tubular vertical post 220 having an aperture or slot 230 in its
side wall. A horizontal arm 240 has one end inside post 220 and secured to
a vertical rod 250 which is suitable mounted so that it can be driven
vertically up and down. The lower end of the vertical rod 250 or an
extension thereof passes through a small motor 260 secured to the lower
surface 32 of the top wall 30. The lower end of the rod 250 carries a
laterally projecting arm 263 which is positioned to operate with an
optical sensor 280 positioned adjacent thereto.
Referring again to the horizontal arm 240, (FIGS. 1, 2, and 6) the outer
end thereof terminates in a small solid cylinder 243 which is oriented
vertically and is provided with two through-holes 247 and 249 which
communicate with a hollow tube 248 which extends downwardly from the solid
cylinder in alignment with the holes 180 in table 170 and the sample cups
therein. The hollow tube 248 is of a small diameter and is dimensioned so
that it can enter a sample cup 190 and extend to about the bottom thereof
to enter fluid therein.
The box 100 contains the same apparatus as box 90 as described above. The
corresponding parts in box 100 carry the same reference numerals as the
parts in the box 90 but primed.
The boxes 90 and 100 include means for applying electrical potential across
the apparatus 10. This means includes a first wire electrode 360 having
one end secured to a power input terminal 363. (FIGS. 1 and 2) in the rear
upwardly in the hollow tube 220 adjacent to the vertical rod 250 and out
of the opening 230 in the side wall and through the hole 249 in cylinder
243 down through the tube 248 to the end thereof so that it can rest in a
fluid in a sample cup when the apparatus 10 is in operation.
The electrical means also includes a similar wire electrode 360' secured to
a power input terminal 365 in the rear wall 50 of the apparatus 10. This
electrode follows a similar path through tube 250' and cylinder 243' into
the tube 248' associated therewith for ultimate insertion into a sample
cup. The electrodes 360 and 360' are preferably of platinum or the like
and are adapted to carry the voltages used in operation of the invention.
A power supply 367 is provided for connection to terminals 363 and 365 to
electrodes 360 and 360' for providing the required voltages. Power supply
367 may also provide whatever other power is needed by the apparatus 10
such as for the motors 210, 210' and 260, 260'. Other auxiliary power
supplies may be provided as desired.
In one embodiment of the invention, illustrated in FIG. 2, the power supply
367 may be of such small size that it can be mounted within support member
20 at any suitable location so that the apparatus 10 has its own
self-contained power supply which may be manually or computer-controlled.
When a built-in power supply is provided, referring to FIG. 7, a voltmeter
376 and an ammeter 378 are secured to the front wall 40 of support member
20 along with a rheostat for adjusting the operating voltage shown on the
voltmeter.
A preferred structure for the vertical posts 250, 250', to insure
electrical safety when high voltage is applied to electrode 360, is shown
in FIG. 8. This embodiment for ease of construction includes an outer post
420 and slidable inner post 422 which are both generally square or
rectangular in construction. The outer post includes a slot 426 in its
side wall and a horizontal arm 428 extends therethrough from the inner
post 422. Arm 428 terminates in cylinder 243. The cable 360 comes up from
the terminal 363 and runs inside the outer post 420 and terminates in a
rigid, relatively large-area flat metallic electrode 430 positioned
perhaps half-way up the post to slightly below the slot 426 therein.
Similarly, the slidable inner post 422 carries on its outer surface a
relatively large area electrode 432 which is suitably positioned so that
when the inner post lowered to operating position, the two electrodes 430
and 432 are in contact with each other. A thin platinum 434 runs from the
electrode 432 through horizontal arm 428 and into the cylinder 243 and
hollow tube 248 as described above.
Referring to FIGS. 1 and 2 an optical detector 290 for use in detecting
material passing through an optical detector 290 for use in detecting
materials passing through a capillary tube which extends through the
detector is seated on a support frame 300 secured to the top wall 30 of
the support member 20 adjacent to the box 100. The apparatus 10 is
designed to use a detector known as an on-column detector of the type
which uses ultraviolet or fluorescent light in the detection process. Such
detectors are made by ISCO of Lincoln, Neb. and EM SCIENCE of Cherry Hill
N.J.
For use with the apparatus of the invention 10, modifications of the
commercial detectors were made in the cuvette | | |
