U.S. patent number 3,836,335 [Application Number 05/366,130] was granted by the patent office on 1974-09-17 for reagent storage and dispensing system.
This patent grant is currently assigned to Corning Glass Works. Invention is credited to William R. Eppes.
United States Patent |
3,836,335 |
Eppes |
September 17, 1974 |
REAGENT STORAGE AND DISPENSING SYSTEM
Abstract
Pressure vessels each have a movable separator which divides the
vessel into two chambers. During filling, reagent is supplied to
one chamber. During a dispensing operation, water pressure is
applied to the other side of the separator. Solenoid valves are
selectively operated to dispense accurate volumes of reagent from
the vessels under pressure supplied by the water on the other side
of the separator.
Inventors: |
Eppes; William R. (Cary,
NC) |
Assignee: |
Corning Glass Works (Corning,
NY)
|
Family
ID: |
23441785 |
Appl.
No.: |
05/366,130 |
Filed: |
June 1, 1973 |
Current U.S.
Class: |
422/50;
210/257.1; 222/386.5; 422/561 |
Current CPC
Class: |
G01F
11/086 (20130101); G01N 35/1002 (20130101) |
Current International
Class: |
G01N
1/00 (20060101); G01F 11/08 (20060101); G01F
11/02 (20060101); G01n 001/10 () |
Field of
Search: |
;23/253,230,259,292
;222/395,386,386.5 ;141/130 ;73/425.4R |
References Cited
[Referenced By]
U.S. Patent Documents
Primary Examiner: Serwin; R. E.
Attorney, Agent or Firm: Zebrowski; Walter S. Patty, Jr.;
Clarence R.
Claims
What is claimed is:
1. A system for storing and dispensing reagents which stain a film
of blood cells deposited on a laboratory slide comprising:
a plurality of pressure vessels each having a movable separator
inside said vessel to divide each vessel into two chambers,
means for supplying reagents to one chamber of each pressure vessel
during filling thereof,
means for applying a regulated fluid pressure to the chamber on the
other side of the movable separator in each pressure vessel,
and
a plurality of solenoid operated valves, one for each pressure
vessel, said solenoid valves being selectively actuated for
intervals of time which dispense accurate volumes of reagent from
said vessels under pressure supplied by the fluid on the other side
of said movable separator.
2. The system recited in claim 1 wherein said separator is a
piston.
3. The system recited in claim 1 wherein said separator is a
flexible, impermeable membrane attached to the inside of said
vessel.
4. The system recited in claim 3 wherein said membrane is attached
around the periphery of the approximate middle of each vessel, and
wherein said membrane is sufficiently flexible to extend almost
completely to one side of said vessel when said vessel is
completely filled with reagent and to the other side of said vessel
when emptied of reagent whereby said reagents are isolated from
materials with which they may react.
5. The system recited in claim 4 further comprising:
a magnet attached to said flexible membrane, and
a reed switch positioned in the chamber containing reagent so that
it is actuated by said magnet when the reagent supply is near
depletion.
6. The system recited in claim 1 wherein there are three pressure
vessels containing a fix, a stain and a buffer, the solenoid
operated valves connected to said vessels being sequentially
operated to apply fix, stain and buffer to said slide.
7. The system recited in claim 1 wherein said means for supplying
reagents comprises:
a flexible supply reservoir containing a reagent, and connections
between said supply reservoir and one of said pressure vessels.
8. The system recited in claim 7 wherein said connections include a
bladder valve connected to said pressure vessel and a probe on said
supply reservoir, said probe being inserted into said bladder valve
during filling of said pressure vessel.
9. The system recited in claim 7 wherein said connections
include:
a plurality of needle fittings, one connected to each of said
solenoid valves,
a plurality of needle-like dispensers connectable to said fitting
for dispensing said accurate volumes of reagent, and
a plurality of mating fittings, one connected to each flexible
supply reservoir and each being connectable to one of said needle
fittings for supplying reagent from said supply reservoir to a
pressure vessel.
10. The system recited in claim 9 wherein each of said mating
fittings includes a valve which permits flow of reagent from said
supply reservoir and which blocks flow of air into said reservoir.
Description
BACKGROUND OF THE INVENTION
This invention relates to reagent storing and dispensing systems
and more particularly to a system for storing and dispensing fix,
stain and buffer reagents for staining a film of blood cells on a
laboratory slide.
In the analysis of blood samples, the blood is smeared on a
laboratory slide and the smear is stained. By counting the
leukocytes on the stained smear, laboratory technicians perform
what is referred to as a white blood cell differential. Automation
of this differential has significant economic impact because the
differential is performed so frequently at every hospital. A thesis
by J. W. Bacus, "An Automated Classification of the Peripheral
Blood Leukocytes by Means of Digital Image Processing," University
of Illinois, Chicago, 1971, describes one automated system.
Copending application Ser. No. 353,004, filed Apr. 20, 1973,
Douglas A. Cotter, "Image Scanning Converter for Automated Slide
Analysis," describes a system developed by my co-workers for
automatically scanning and digitizing the count of the leukocytes
on the stained smear.
SUMMARY OF THE INVENTION
In accordance with this invention a reagent storing and dispensing
system includes a plurality of pressure vessels each having a
movable separator which divides the vessel into two chambers.
Reagent is supplied to and stored in one of the chambers. A
regulated water pressure is applied to the other chamber. solenoid
operated valves are selectively actuated for intervals of time
which dispense accurate volumes of reagent from the vessels under
pressure supplied by the water pressure against the separator. In
this manner, accurate volumes of reagent are supplied and the time
at which they are delivered is easily adjustable. Furthermore, the
pressure vessel of this invention effectively isolates the reagent
from moisture which can be absorbed from the air or from any other
material with which the reagent may react.
Further in accordance with this invention the pressure vessels are
filled from flexible supply reservoirs. In one embodiment of the
invention the connection between the resevoir and the pressure
vessel includes a needle fitting connected to the solenoid valve
and a mating fitting connected to the reservoir. After the pressure
vessel is filled, a needle-like dispenser is placed on the needle
fitting. The needle-like dispenser supplies accurate volumes of
reagent passed by the associated solenoid valve.
The foregoing and other objects, features and advantages of the
invention will be better understood from the following more
detailed description and appended claims.
DESCRIPTION OF THE DRAWINGS
FIGS. 1A and 1B together show the reagent storage and supply
system;
FIG. 2 shows the supply reservoir;
FIG. 3 is a cross-section of the pressure vessel;
FIG. 4 is a view of the outside of the pressure vessel;
FIG. 5 shows a modification of the pressure vessel;
FIG. 6 shows a modified connection of the flexible supply
reservoir; and
FIG. 7 shows the mating fitting of FIG. 6.
DESCRIPTION OF THE PREFERRED EMBODIMENT
In FIGS. 1A and 1B the system is shown in the pressurized stand-by
condition. Water at line pressure passes through the pressure
regulator 10 where pressure is reduced to approximately 8 to 10
lbs. per square inch. (PSIG. Part of the available water is applied
to solenoid valve 16. (The solenoid valve 16 is depicted in its
closed condition. In the notation used two states of the valve are
depicted by two blocks. The lower block depicts the closed
condition whereas the upper block depicts the open condition of the
valve.) When the solenoid valve 16 is actuated, available water
passes through the valve 16 to the rinse discharge tube 18.
Water at 8 PSI is also applied to the regulator 12 which reduces
the pressure to approximately 5 PSIG. Water at this pressure passes
through the solenoid valve 14 into the water containing chambers of
the pressure vessels 22, 24 and 26. Each of the pressure vessels
are divided into two chambers by a separator as will be described
more fully with respect to FIGS. 3 and 5.
In the staining of blood smeared laboratory slides, pressure vessel
26 will normally contain a fix, such as Wright stain, in the
reagent containing chamber. Pressure vessel 24 contains a stain
such as Giesma stain and pressure vessel 22 contains a buffer in
the reagent chamber. Lines on the reagent side of each pressure
chamber are connected to solenoid operated valves 28, 30 and 32
which dispense reagent from the pressure chambers 22, 24 and 26
respectively.
These valves are connected to dispensers 40, 42 and 44 which
sequentially apply the reagents to the slide. The dispensers 40, 42
and 44 are of an appropriate diameter which typically is 0.023
inches. They are individually removable for ease of cleaning or for
replacing with dispensers of a different diameter to achieve
different flow rates.
The solenoid valves 28, 30 and 32 control when and how much reagent
is dispensed. The time in the process cycle at which the valve is
energized determines when the liquid is dispensed. The length of
time that the valve is energized determines the volume which is
dispensed.
Liquid from the rinse process is collected in the drain pan 46 and
carried away by the drain line 48.
When it is desired to refill the pressure vessels 22, 24 and 26
with reagent, the solenoid valve 14 is actuated to provide a
connection between the bottom chambers of the pressure vessels and
a pump 15. The pump 15 is wired in parallel with the solenoid valve
14. When a fill push button, not shown, is depressed both the
solenoid valve 14 and the pump 15 are energized. Water is pumped
from the bottom chambers of the vessels 22, 24 and 26 and into the
drain pan 46. In one embodiment of the invention new reagent is
supplied to each of the vessels during a fill operation through
bladder valves 34, 36 and 38. Bladder valves 34, 36 and 38 are
respectively connected to the pressure vessels 22, 24 and 26. These
bladder valves are part of a connection between a supply reservoir
53 containing the appropriate reagent and the pressure vessel in
which this reagent is to be stored. In FIG. 1A the connection
between the supply reservoir and the pressure vessel is shown as
including a valve 15A and a mating fitting 17 which, in the first
embodiment to be described is a probe.
The supply reservoir of this first embodiment is shown in more
detail in FIG. 2. It includes a flexible collapsible container 53.
Typically this is constructed of three mil polyethylene. A line
from the supply reservoir leads to a check valve 51 and thence to a
dispensing probe 52. When the probe 52 is inserted into a bladder
valve 34, 36 or 38 reagent from the supply reservoir flows into the
pressure vessel.
A first embodiment of the pressure vessel is shown in FIGS. 3 and
4. It includes the flexible membrane 61. One type of suitable
material for this impermeable membrane is butal rubber on a dacron
matrix. The membrane is attached around the periphery of the
approximate middle of the vessel. The membrane is sufficiently
flexible to extend almost completely to one side of the vessel. As
shown in FIG. 3, the membrane extends to the bottom of the vessel
when it is filled with reagent. As the reagent is emptied from the
vessel the membrane 61 follows the reagent and isolates it from air
or other material with which the reagent might react. In
particular, those reagents containing methyl alcohol may otherwise
absorb moisture from the air. The membrane 61 is sufficiently
flexible to extend completely to the top of the vessel when emptied
of reagent.
Water enters the pressure vessel through the inlet 62 and reagent
leaves the vessel through the outlet 63. The pressure vessel
includes a bottom portion 64 and a top portion 65 which are secured
together with the diaphragm between them. The lower portion 64 and
the upper portion 65 of the pressure vessel are constructed of a
nonmagnetic material such as acetal copolymer.
A warning system informs the operator when the reagent supply is
near depletion. A magnet 67 is attached to the membrane. As the
reagent is depleted the membrane is forced upward until the magnet
67 approaches the reed switch 66 which is attached to the vessel
wall. When the magnet is sufficiently close, the switch closes
completing the circuit to an indicator light 68 (FIG. 4) which is
mounted in a suitably visible location on the machine. The operator
is advised of a nearly depleted reagent.
The operation of the system is described with reference to FIGS. 1A
and 1B. The procedure for filling a dry system is as follows. The
line leading to the regulators 10 and 12 is pressurized. The valve
14 is in the position shown. Water pressure of approximately 8 PSI
is applied to solenoid valve 16 and water pressure of approximately
5 PSIG is applied to the water containing chambers of pressure
vessels 22, 24 and 26. The flexible membrane in each pressure
vessel is deformed and either compresses the air on the opposite
side of the membrane to 5 PSIG or fills the available space in the
chamber, whichever occurs first. The solenoid valves 28, 30 and 32
are momentarily opened by a manual override. Any trapped air is
bled out insuring that all membranes in the pressure vessels are
forced completely to the reagent side of the pressure vessel.
The procedure from this point on is the procedure used to fill a
previously charged system. Valve 14 and pump 15 are energized. The
fitting 17 on the supply reservoir 53 is fitted to the proper
bladder valve.
Because water is being drawn from the water chamber of the pressure
vessel, flow of reagent is induced from the supply reservoir 53,
through fitting 17 and a bladder valve (34, 36 and 38) into the
reagent chamber of the pressure vessel. Flow will continue until
the pressure vessel is completely filled with reagent or the valve
14 and pump 15 are de-energized.
It would seem possible to fill the pressure vessels by applying a
positive pressure directly to the flexible supply reservoir by
squeezing it. However, in this case it is possible to rupture the
supply reservoir by applying too great a pressure in this manner.
The arrangement by which reagent is drawn out of the supply
reservoir is preferred.
When the system has been filled the probe or probes are
disconnected from their respective bladder valves. Solenoid valves
28, 30 and 32 are momentarily opened to bleed any air that may be
trapped in the reagent side of the system. The system is now in the
dispensing mode and is ready for use. Since the pressure vessels
22, 24 and 26 are all subjected to 5 PSIG, energization of solenoid
valves 28, 30 or 32 will allow the associated reagent to flow. The
pressure in the system, the viscosity of the reagent and the time
the solenoid valve is energized are all closely controlled.
Therefore, the volume of reagent dispensed is repeatable within 10
percent.
A modification of the pressure vessel is shown in FIG. 5. The
pressure vessel has a separator which includes a piston 70 and a
flexible boot 71. This allows the piston 70 to travel all the way
to the bottom of the pressure vessel when it is filled with reagent
or all the way to the top of the vessel when the reagent is
depleted. The warning system for warning the operator when the
reagent supply is depleted includes a reed switch 72, a magnet 73
and a shunt 74. When there is reagent in the vessel, the magnet 73
is shunted by the shunt 74. However, when the reagent supply is
depleted the shunt 74 is lifted off the magnet 73. The magnet 73
energizes the reed switch 72 thereby activating the alarm. When the
vessel is refilled the shunt 74, which is nickel plated mild steel,
contacts the magnet 73 thereby diverting its field away from the
reed switch. The reed switch opens and turns off the "low reagent"
alarm.
A modification of the connection between the reagent reservoir and
the pressure vessel is shown in FIGS. 6 and 7. In this modification
the bladder valves 34, 36 and 38 and their connections to the
pressure vessels are eliminated. The dispensers 40, 42 and 44 of
FIG. 1B are modified so that they are used both for dispensing
reagent and for refilling the the pressure vessel. As shown in FIG.
6, each dispenser includes a male Luer needle fitting 75. During
dispensing, a needle 76 is attached to the needle fitting 75. The
needle 76 may be a standard medical needle which provides the
desired flow rate of staining reagent to the slide. Use of such a
needle has the advantage that it can be periodically replaced;
otherwise dried reagent tends to change the size of the opening in
the dispenser changing the flow rate.
When it is desired to refill a pressure vessel, a mating female
Luer fitting 77 is placed on the needle fitting 75. This fitting
may include a simple check valve. Or, as shown in FIG. 7 an
alternate action valve actuator 78 may be provided to actuate a
valve. The valve actuator is pushed one way to open the valve
connection between the reservoir and the fitting. It is pushed the
other way to close the valve and prevent air from entering the
reservoir.
The operation of the modified system is as follows.
The needle 76 is removed from the needle fitting 75 connected to
the pressure vessel to be refilled. This assures a maximum flow and
hence a minimum filling time. The female Luer fitting 77 is
connected to the needle fitting 75. The valve actuator 78 is
operated to set the valve in the open position. The "fill" push
button is depressed. This energizes valve 14 and pump 15 (FIG. 1A).
When the pressure vessel is filled, the valve actuator 78 is
depressed to set the valve in a closed position. The fitting 77 is
disconnected from the fitting 75. The needle 76 is reattached to
the fitting 75 and the system is again ready for dispensing.
While a particular embodiment of the invention has been shown and
described, various modifications are within the true spirit and
scope of the invention. The appended claims are intended to cover
any such modifications.
* * * * *