Apparatus for performing binder-substrate processing of clinical substances is disclosed. An operating head is provided which includes an array of upwardly opening receptacles each having a closed bottom. A temperature control element is located in the operating head proximate the receptacles. The operating head also includes a manifold having an influx chamber, an efflux chamber and a plurality of hollow prongs emanating from the influx chamber and through the efflux chamber. The prongs extend downwardly into respective of the receptacles to provide fluid communication between the influx chamber and the interior of the receptacles through the prongs. The efflux chamber is in fluid communication with the open ends of the receptacles to drain fluid therefrom. A solid phase binder is located in the bottoms of the respective receptacles, and may be coated on solid phase elements located in the receptacles or on the receptacles themselves. The solid phase binder is immersed in the clinical substances for incubation by the temperature control element. After incubation of the clinical substances, the operating head is inverted. A washing fluid is injected into the influx chamber of the inverted operating head so that the washing fluid flows upwardly through the prongs to wash excess of the clinical substances from the solid phase binder retained in the receptacles. The washing fluid is drained from the operating head through the efflux chamber. A drying fluid is usually injected into the influx chamber of the inverted operating head so that the drying fluid flows upwardly through the prongs and into the receptacles to dry the solid phase binder and incubated clinical substances adhering thereto.

BACKGROUND OF THE INVENTION 
The present invention relates to apparatus for performing the incubation 
and washing requirements of solid-phase, binder-substrate analysis of 
clinical substances. 
A larger number of different types of tests are performed on various 
clinical substances such as human serum by using binder-substrate 
analysis. In such an analysis, a binding agent is typically suspended in a 
suppportive medium to which might be added either a quantitated standard 
substrate or the unquantitated substrate in the clinical substance. 
Additionally, a measure of labeled substrate, as nearly identical to the 
substrate of the standard and clinical substance as possible, is added to 
the medium to serve as a marker for the distribution of all substrate 
during incubation. This supportive medium is then incubated under 
precisely controlled conditions favoring the equilibrium of 
binder-substrate complex and the individual components. After the 
incubation has achieved a point of equilibrium, some method of separation 
is performed allowing isolation of either the binder-substrate complex or 
the free components. 
Since the labeled substrate is selected for those pertinent similarities to 
the standard and unquantitated substrate, it will distribute itself 
exactly like those unlabeled substrates at equilibrium. Measurement by 
suitable means of that label in the isolated fraction will provide the 
same results as if the total (labeled and unlabeled) substrate in the 
isolated fraction is measured. Comparison of the amount of substrate found 
in the isolate of the unquantitated clinical substances to that of the 
standards provides the result of the analysis. 
One such method of separation, isolating the binder-substrate complex from 
the components, is termed "solid phase". Solid phase defines the physical 
condition of the binder-substrate complex at the point of equilibrium when 
the separation step is to occur. Typically, the binder is originally the 
solid phase component. When incubation is terminated, the separation step 
involves isolation of the solid phase binder-substrate complex from the 
liquid phase of the assay medium. If the binder is physically attached to 
microscopic-solid particles, the separation would involve a centrifuge. If 
the binder is attached to a large enough particle such as a 1/4 inch dia. 
sphere or the bottom interior surface of a test tube, the separation step 
involves decantation or aspiration of the liquid phase from the 
immobilized, solid-phase binder-substrate complex. 
The application of solid phase to the binder-substrate analysis has 
eliminated the centrifuge in several cases (where large enough supports 
are used); but added to the procedure, time and/or temperature enhancement 
of the incubation step, and a wash step(s) to adequately isolate the solid 
phase complex from the liquid phase components. These additional 
requirements must be controlled to guarantee reproducibility between all 
specimens. 
One such application of solid-phase binder-substrate analysis determines 
the presence of hepatitis virus in serum specimen. The primary binder is 
an antibody to hepatitis virus, attached to a 1/4 inch dia. bead. This 
solid-phase binder is immersed in a buffered medium containing a measure 
of specimen, all contained within a test tube. 
The test tube is maintained at a controlled temperature for sufficient time 
(incubation) to allow optimum formation of the antibody-virus complex on 
the surface of the plastic bead. The liquid medium containing all 
unreacted substances is then removed from the test tube and the bead and 
inner surfaces of the test tube are adequately washed leaving the solid 
phase antibody-virus complex. A second antibody, similar in binding 
properties to the primary but solubilized in a buffered medium, is then 
added to the test tube containing the solid phase complex. Additionally 
the second antibody is suitably labeled to allow subsequent detection. The 
test tube is again maintained according to those parameters optimizing the 
formation of a second binding; the virus by the labeled antibody. The only 
virus available to this second, labeled antibody is carried over through 
the washing(s) from the first incubation. Therefore, any secondary binding 
will join the labeled antibody to the solid-phase antibody-virus complex 
forming a sandwich. The liquid medium containing all unreacted substances 
is again removed from the test tube and the bead and inner surfaces of the 
test tube are washed leaving the solid-phase primary 
antibody-virus-labeled antibody complex. The plastic bead is then 
transfered to a carrier-tube suitable to the label detection apparatus. 
Detection of label on the plastic bead implies presence of virus since no 
other form of labeled antibody is carried over through the washing(s) from 
the second incubation. 
At present, the incubations, washings and drying of the solid phase 
components in this variety of binder-substrate analysis are generally 
performed by hand. The test consumes a large amount of technician time, 
thus labor costs are relatively high. The large number of manual steps 
often cause errors in the testing procedures. Variations in the parameters 
of incubation time and temperature, and wash volumes, inherent in manual 
processing, produces nonuniformity in test results. A further problem 
presented by extensive manual manipulation is cross-contamination of the 
samples and the constant exposure of the technician to potential 
biological and radioactive hazards. 
SUMMARY OF THE INVENTION 
The present invention provides apparatus for performing binder-substrate 
processing of clinical substances. An operating head is provided which 
includes an array of upwardly opening receptacles each having a closed 
bottom. A temperature control element is located in the operating head 
proximate the receptacles. The operating head also includes a manifold 
having an influx chamber, an efflux chamber and a plurality of hollow 
prongs emanating from the influx chamber and through the efflux chamber. 
The prongs extend downwardly into respective of the receptacles to provide 
fluid communication between the influx chamber and the interior of the 
receptacles to the prongs. The efflux chamber is in fluid communication 
with the open ends of the receptacles to drain fluids therefrom. A solid 
phase binder is located in the bottoms of the respective receptacles, and 
may be coated on solid phase elements located in the receptacles or on the 
receptacles themselves. The solid phase binder is immersed in the clinical 
substances for incubation by the temperature control element. After 
incubation of the clinical substances, the operating head is inverted. A 
washing fluid is injected into the influx chamber of the inverted 
operating head so that the washing fluid flows upwardly through the prongs 
to wash excess of the clinical substances from the solid phase elements in 
the receptacles. The washing fluid is drained from the operating head 
through the efflux chamber. A drying fluid is often injected into the 
influx chamber of the inverted operating head so that the drying fluid 
flows upwardly through the prongs and into the receptacles to dry the 
solid phase elements and incubated clinical substances adhering thereto. 
The present invention eliminates a large number of the manual steps which 
have been required in the past to perform binder-substrate processing. The 
time required of a laboratory technician to perform this type of analysis 
is substantially reduced, resulting in a corresponding reduction in labor 
costs. Furthermore, the analysis can be performed in a repeatable and 
consistent manner, giving greater uniformity in the results. The 
orientation and relative positioning of the test tubes during washing 
virtually eliminates the potential for cross-contamination of samples. The 
overall design of the unit allowing isolation of both liquids and vapors 
eliminates the hazards to the technician. In addition, a large number of 
individual samples can be processed at the same time, under virtually 
identical conditions, further increasing the efficiency of the system. 
The novel features which are characteristic of the invention, as to 
organization and method of operation, together with further objects and 
advantages thereof will be better understood from the following 
description considered in connection with the accompanied drawings which a 
preferred embodiment of the invention is illustrated by way of example. It 
is to be expressly understood, however, that the drawings are for the 
purpose of illustration and description only and are not intended as a 
definition of the limits of the invention.

DESCRIPTION OF THE PREFERRED EMBODIMENT 
The construction of the preferred embodiment of the binder-substrate 
processor 10 of the present invention is illustrated primarily by way of 
reference to FIGS. 1 and 2. Processor 10 includes a control unit 12 having 
an operating head 14 superimposed thereabove. Operating head 14 is mounted 
on shafts 15, 16. Shaft 16 is driven by gears 17, 18 and inverting arm 20 
to control the orientation of operating head 14. A rocker arm 21 also 
connects the shaft 16 to rock head 14 for purposes described below. 
Operating head 14 includes a base 22 having an internal heating block 26 
controlled by switch 23 and thermostat 24. A plurality of recesses 28 are 
formed in heating block 26. A rack 30 supporting a plurality of test tubes 
32 is attachable to base 22 (see FIG. 2). When rack 30 is so attached, 
test tubes 32 extend downwardly into recesses 28 in heating block 26 so 
that the heating block surrounds the bottoms of the test tubes. A solid 
phase element such as bead 33 coated with a solid phase binder may be 
located at the bottom of each test tube 32. Alternatively, the solid phase 
binder may simply be coated on test tube 32 which then acts as the solid 
phase element or on a plurality of solid phase elements located in the 
test tubes. 
A cover 34 fits over and attaches to base 22 to form the upper portion of 
the operating head. A rib 36 on the underside of cover 34 fits within a 
corresponding groove 38 to form a water tight seal between the cover 34 
and base 22. Cover 34 is located in position by wing nut 40 which engages 
slots 42 projecting outwardly from cover 34 (see FIG. 1). 
When cover 34 is fixed to base 22, a chamber is formed between a plate 44 
within cover 34 and the upper portion of base 22, which acts as an efflux 
chamber. In addition, plate 44 defines a chamber within cover 34 which is 
designated the influx chamber. A plurality of hollow prongs 46 extend 
downwardly from plate 44 through the efflux chamber and into the interior 
of test tubes 32. Prongs 46 are in fluid communication with the influx 
chamber and confine each bead 33, if used, in the bottom of its respective 
test tube 32. The influx and efflux chambers together with prongs 46 
provide a manifold system for operating head 14, as described in detail 
hereinafter. 
A container 48 of washing fluid is connected with control unit 12 by tube 
50. The washing fluid is maintained under pressure in container 48, and a 
relief valve 52 is provided to relieve excess pressure. Another tube 54 
emanates from control unit 12 and connects with a source of pressurized 
air or another drying fluid (not shown in FIG. 1). A plurality of tubes 56 
interconnect control unit 12 with operating head 14 as will be described 
in more detail hereinafter. Various switches 58 and timers 60 are provided 
to control the operation of processor 10. 
The function of the various fluid connections in the analyzer of the 
present invention are illustrated in more detail by way of reference to 
the schematic view of FIG. 3. In FIG. 3, operating head 14 is illustrated 
in its inverted position, which is accomplished by rotating operating head 
14 on shafts 15, 16 (see FIGS. 1 and 2). In this configuration, the base 
20 of operating head 14 is located above cover 34. Each test tube 32 (only 
one test tube is illustrated for clarity) now opens downwardly, and each 
prong 46 projects upwardly into each respective test tube. Efflux chamber 
60 is now superimposed over influx chamber 62. Operating head 14 is shown 
in its inverted configuration in FIG. 3 because all fluids are applied 
while the operating head is so inverted. 
Container 48 of wash solution 64 is communicated to control unit 12 through 
tube 50. A valve 66 is located within the control box 12, and is operated 
by a solenoid 67 to control the flow of washing fluid to influx chamber 62 
through tube 68. When valve 66 is open, washing fluid is injected into 
influx chamber 62, and passes upwardly through prong 46 into receptacle 
32. At the same time, valve 73 is opened by solenoid 75 so that the 
washing fluid can drain out of efflux chamber 60 through tube 72 for safe 
disposal. 
A source 74 of compressed air or other drying medium is connected to 
control unit 12 by tube 54. A pressure regulator 78 is located 
intermediate tube 54. A valve 80 actuated by solenoid 81 controls the flow 
of the compressed air under pressure to influx chamber 62 through tube 82. 
When the valve 80 is open, compressed air is injected into influx chamber 
62, and passes upwardly through prong 46 and into receptacle 32. Valve 70 
is opened at the same time to allow air to escape through efflux chamber 
60 and tube 72. 
Compressed air 74 is also communicated to container 48 of wash solution 64 
through tube 84 to maintain the wash solution under pressure. In addition, 
valve 86 may be opened by actuating solenoid 87 to communicate compressed 
air to efflux chamber 60 through tube 88 to blow the wash solution out of 
the chamber and prevent a vacuum from forming in the efflux chamber during 
certain draining operations. 
An exemplary manner in which a binder-substrate processor is performed by 
processor 10 is illustrated by way of reference to the schematic views of 
FIGS. 4A-4F. In each of these figures, operating head 14 is illustrated 
schematically together with a pair of test tubes 32 and their associated 
apparatus. Valves 66, 70, 73, 80 and 86 correspond with valves denoted by 
the same numerals in FIG. 3. 
Referring initially to FIG. 4A, operating head 14 is in its normal upright 
position, and test tubes 32 open upwardly. Beads 33 are located in the 
bottom of test tubes 32 as illustrated in FIG. 4A, and are immersed in a 
diluted sample 92. Typically, beads 33 are plastic and coated with a 
binder such as an antibody to a virus or other suspected constituent of 
sample 92. Often, beads 33 are not used and the binder is coated directly 
on the inside walls of test tubes 32. Heating block 26 is actuated to 
incubate sample 92 and generate reactions which may be possible between 
certain constituents of the diluted sample and the binder. Operating head 
14 may be rocked back and forth on shafts 15, 16 using rocker arm 21 (see 
FIGS. 1 and 2) during the incubation period to facilitate any such 
reaction. During the incubation phase, all valves 66, 70, 73, 80 and 86 
are generally closed. 
After incubation, operating head 14 is inverted as illustrated in FIG. 4B 
by rotating it on shafts 15, 16 (see FIGS. 1 and 2). The dimensions of 
beads 33 are larger than the annular distance between prongs 46 and test 
tubes 32 so that the beads are confined within the test tubes. Excess 
diluted sample 92 which has not adhered to the binder on beads 33 is 
drained from test tubes 32. If the sought after virus was contained in the 
patient sample, a certain quantity thereof will be bound to the binder on 
beads 33. 
The binder on beads 33 is then washed as illustrated by way of reference to 
FIG. 4C. Valve 66 is opened, and washing fluid 64 is injected into influx 
chamber 62 as illustrated by arrows 94. Washing fluid 64 passes upwardly 
through prongs 46 to wash the binder on beads 33 and the interior of test 
tubes 32. The washing fluid then drains through test tubes 32 along the 
outside of prongs 46 and into efflux chamber 60. Valve 70 is also open so 
that washing fluid 64 can drain from efflux chamber 60 through tube 72, as 
depicted by arrows 96. 
After the wash cycle is completed, valve 66 is closed and valve 86 is 
opened as illustrated in FIG. 4D. Compressed air passes into efflux 
chamber 60 through tube 88 as depicted by arrow 98 to allow the wash 
solution to drain through tube 72. 
After the wash fluid has been drained from efflux chamber 60, valve 73 is 
opened as depicted in FIG. 4E to allow the wash fluid in influx chamber 62 
to be drained from the system through tube 69. Valve 70 may be closed 
during this operation but valve 86 remains open to blow out the remaining 
wash solution and to prevent a vacuum from forming in the system. 
After all of the wash solution has been drained from operating head 14, the 
binder on beads 33 and any clinical substances bound thereto are dried as 
illustrated in FIG. 4F. Valve 86 is closed, and compressed air or another 
drying medium is injected into influx chamber 62 through tube 88 by 
opening valve 80. The compressed air passes upwardly through prongs 46 to 
completely dry beads 33 and substances bound thereto and the interior of 
test tubes 32. The drying air passes out of test tubes 32 into efflux 
chamber 60 and out of operating head 14 through tube 72. 
To operate substrate analyzer 10, test tubes 32 containing a binder coated 
to the interior of the test tubes or to a plurality of beads 33 located in 
the individual test tubes are placed in rack 30. Various dilute samples to 
be tested are then dispensed into respective test tubes, and rack 30 is 
mounted to the base 22 of operating head 14. Cover 34 is then placed over 
the operating head and secured thereto. 
The parameters of the incubation, wash, and dry cycles are set as desired 
on processor 10 using the various dials 24 and 60 provided for that 
purpose. Processor 10 is then actuated, and automatically performs the 
incubation, wash, and dry cycles as preselected. After the cycles have 
been completed, it may be desirable for certain types of tests to add a 
second clinical substance to the various test tubes and the entire process 
can be repeated to perform the second phase of such tests. 
While a preferred embodiment of the present invention has been illustrated 
in detail, it is apparent that modifications and adaptations of that 
embodiment will occur to those skilled in the art. However, it is to be 
expressly understood that such modifications and adaptations are within 
the spirit and scope of the present invention, as set forth in the 
following claims: