U.S. patent number 3,791,517 [Application Number 05/338,216] was granted by the patent office on 1974-02-12 for digital fluidic amplifier particle sorter.
This patent grant is currently assigned to Bio/Physics Systems, Inc.. Invention is credited to Mitchell Friedman.
United States Patent |
3,791,517 |
Friedman |
February 12, 1974 |
**Please see images for:
( Certificate of Correction ) ** |
DIGITAL FLUIDIC AMPLIFIER PARTICLE SORTER
Abstract
Differences in small particles entrained in a stream of liquid
are detected and the resultant difference signals are used to
control a fluidic amplifier located downstream to switch the liquid
particle carrying stream to different outlet ports determined by
the detection of particle differences.
Inventors: |
Friedman; Mitchell (Yorktown
Heights, NY) |
Assignee: |
Bio/Physics Systems, Inc.
(Mahopac, NY)
|
Family
ID: |
23323903 |
Appl.
No.: |
05/338,216 |
Filed: |
March 5, 1973 |
Current U.S.
Class: |
209/579; 209/546;
209/563; 209/606; 209/644; 209/906; 324/71.4; 356/39 |
Current CPC
Class: |
B07C
5/36 (20130101); B07C 5/342 (20130101); F15C
1/08 (20130101); F15C 1/008 (20130101); Y10S
209/906 (20130101); G01N 2015/149 (20130101) |
Current International
Class: |
B07C
5/36 (20060101); B07C 5/342 (20060101); F15C
1/08 (20060101); F15C 1/00 (20060101); G01N
15/14 (20060101); B07c 005/00 () |
Field of
Search: |
;209/3,4,74,111.5,111.6,111.7,111.8 ;324/34,61,71CP ;356/39 ;210/65
;250/222CP |
References Cited
[Referenced By]
U.S. Patent Documents
Primary Examiner: Knowles; Allen N.
Assistant Examiner: Church; Gene A.
Attorney, Agent or Firm: Ailes; Curtis
Claims
I claim:
1. Apparatus for sorting small particles such as biological cells
while the particles are suspended in a liquid comprising
a housing defining an optical chamber and comprised of a material
which transmits light,
means for moving the particle suspending liquid through said
housing in a thin narrow stream to convey the particles in sequence
through the stream one by one,
a light source for directing a beam of light into said housing to
intersect with the stream of particles,
at least one photoresponsive pick-up element positioned to detect
at least one optical reaction of each particle to illumination from
the beam,
said photoresponsive pick-up element being operable to provide
electrical signals which vary in accordance with said optical
reaction of each particle to thereby provide electrical signals
indicative of different particle characteristics,
a digital fluidic amplifier having an inlet connected to the outlet
of said chamber to receive the particle stream,
said fluidic amplifier having a switching chamber communicating
with said inlet and at least two different outlets communicating
with said switching chamber,
an electrical transducer coupled to receive electrical signals from
said photoresponsive element and operable to provide control
signals to said fluidic amplifier,
said fluidic amplifier being operable in response to said control
signals to switch the liquid particle-carrying stream entering the
inlet thereof from one outlet to another selected outlet in
response to a predetermined particle characteristic control signal
from said transducer.
2. Apparatus for sorting small particles such as biological cells
while the particles are suspended in a fluid comprising
a housing defining a detection chamber,
means for moving the particle suspending fluid through said housing
in a stream to convey the particles through the stream,
a detection means associated with said detection chamber for
detecting differences in particle characteristics and operable to
provide electrical signals which vary in accordance with said
differences in particle characteristics,
a digital fluidic amplifier having an inlet connected to the outlet
of said chamber to receive the particle stream,
said fluidic amplifier having a switching chamber communicating
with said inlet and at least two different outlet ports
communicating with said switching chamber,
an electrical transducer coupled to receive electrical signals from
said detection means and operable to provide control signals to
said fluidic amplifier,
said fluidic amplifier being operable in response to said control
signals to switch the fluid particle-carrying stream entering the
inlet thereof from a first outlet port to a second selected outlet
port in response to a predetermined particle characteristic control
signal from said tranducer.
3. Apparatus as claimed in claim 1 wherein;
said housing defines said detection chamber and said fluidic
amplifier in a unitary structure.
4. Apparatus as claimed in claim 1 wherein;
said digital fluidic amplifier is a wall attachment effect
device.
5. Apparatus as claimed in claim 1 wherein;
said housing defining a detection chamber is operable to provide
for a laminar flow of the particle-suspending fluid
therethrough,
said apparatus including means associated with said digital fluidic
amplifier for converting the laminar flow to turbulent flow within
said fluidic amplifier switching chamber.
6. Apparatus as claimed in claim 5 wherein;
said means for converting the laminar flow to turbulent flow
comprises an electrically energized turbulence transducer
communicating with said fluidic amplifier switching chamber.
7. Apparatus as claimed in claim 6 wherein;
said electrical transducer coupled to receive electrical signals
from said detection means is combined in said electrically
energized turbulence transducer.
8. Apparatus as claimed in claim 7 wherein;
said combined transducer is comprised of a piezo-electric
crystal.
9. Apparatus as claimed in claim 7 wherein;
said fluidic amplifier is operable when the fluid entering the
inlet thereof continues in the laminar flow mode to deliver the
fluid through said switching chamber to said first one of said
fluidic amplifier outlet ports,
said electrical transducer coupled to receive electrical signals
from said detection means being combined in said turbulence
transducer and the control signals provided to said fluidic
amplifier comprising turbulence induction signals for selectively
inducing turbulence in the stream of fluid,
said fluidic amplifier being operable in response to the conversion
of the stream of fluid from laminar flow to turbulent flow to
switch the stream in said switching chamber from said first outlet
port to said second outlet port.
10. Apparatus as claimed in claim 9 wherein;
said first outlet port is in axial alignment with the inlet of said
switching chamber and said second outlet port is displaced from a
position of axial alignment with said inlet,
said switching chamber being partially defined by two
sidewalls,
one of said sidewalls being more closely spaced than the other to
the extended axis of the inlet to said switching chamber,
said closely spaced sidewall being arranged on the same side of
said switching chamber as said second outlet port to lead the
stream of fluid to said second port by the wall attachment effect
when said fluid is turbulent.
11. Apparatus as claimed in claim 10 wherein;
said fluidic amplifier includes at least one control port
communicating with one side of said switching chamber near the
inlet thereof,
means for changing the fluid pressure at said control port to be
different from the pressure within said switching chamber to cause
a transfer of fluid between said control port and said switching
chamber to thereby interrupt the wall attachment effect by means of
which the stream of fluid is attached to said closely spaced
wall.
12. Apparatus as claimed in claim 11 wherein;
said fluidic amplifier includes a third outlet port communicating
with the said switching chamber and arranged adjacent to the
sidewall which is not spaced closely to the extended axis of the
inlet to said switching chamber,
said fluidic amplifier being operable to direct the stream of fluid
to said third outlet port by the wall attachment effect on said
last-mentioned sidewall in response to a fluid pressure change
switching signal from said control port and in the presence of
turbulent fluid flow.
13. Apparatus as claimed in claim 5 wherein;
said digital fluidic amplifier includes two control ports
communicating with said switching chamber,
separate valve control means associated with each of said control
ports,
a source of control fluid at a pressure different from the pressure
within said switching chamber connected to each of said valve
means,
each of said control ports being associated with one diverging side
wall of said switching chamber,
each of said control ports being operable in conjunction with the
associated valve means to control the wall attachment effect of the
stream of fluid within said switching chamber with respect to the
associated side wall,
said two outlet ports being respectively associated with said
divergent side walls,
at least one of said valve means being combined in said electrical
transducer.
14. Apparatus as claimed in claim 13 wherein;
said means for converting the laminar flow to turbulent flow
comprises an electrically energized turbulence transducer
communicating with said fluidic amplifier switching chamber.
15. Apparatus as claimed in claim 14 wherein;
said turbulence transducer is operable in conjunction with said
valve means.
16. Apparatus as claimed in claim 14 wherein there is provided,
a third outlet port in axial alignment with said inlet of said
switching chamber,
said third outlet port being operable to receive the stream of
fluid when the fluid flow remains laminar.
17. Apparatus as claimed in claim 16 wherein;
said digital fluidic amplifier is capable of selectively directing
said particle bearing stream to any one of said outlet ports,
said electrically energized turbulence transducer and said separate
valve control means each being connected to receive signals from
said detection means,
one of said valve means being operable upon reception of a signal
in conjunction with reception of a signal by said turbulence
transducer to switch the stream to said first outlet port,
the other one of said valve means being operable upon reception of
a signal in conjunction with reception of a signal by said
turbulence transducer to switch the stream to said second outlet
port.
18. Apparatus as claimed in claim 2 wherein;
said stream of particle carrying fluid is a liquid.
19. Apparatus as claimed in claim 2 wherein there is provided,
at least one additional digital fluidic amplifier having an inlet
connected to one of said outlet ports of said first-mentioned
digital fluidic amplifier,
said additional digital fluidic amplifier having a second
electrical transducer coupled to receive electrical signals from
said detection means and being operable to provide control signals
to said additional fluidic amplifier,
said additional fluidic amplifier being operable in response to
said control signals to switch the fluid particle carrying stream
entering the inlet thereof from a first outlet port to a second
selected outlet port in response to a predetermined particle
characteristic control signal from said second transducer.
20. Apparatus as claimed in claim 2 wherein there is provided,
a second housing defining a second detection chamber connected to
receive the particle suspending fluid from one of said outlet ports
of said fluidic amplifier,
a detection means associated with said second detection chamber for
detecting differences in particle characteristics and operable to
provide electrical signals which vary in accordance with said
differences in particle characteristics,
a second digital fluidic amplifier having an inlet connected to the
outlet of said second chamber to receive the particle stream,
said second fluidic amplifier having a switching chamber
communicating with said inlet and at least two different outlet
ports communicating with said switching chamber,
a second electrical transducer coupled to receive electrical
signals from said second detection means and operable to provide
control signals to said second fluidic amplifier,
said second fluidic amplifier being operable in response to said
control signals from said second detection means to switch the
fluid particle carrying stream entering the inlet thereof from a
first outlet port to a second selected outlet port in response to a
predetermined particle characteristic control signal from said
second transducer.
Description
This invention relates to apparatus for sorting small particles
such as biological cells which may be microscopic in size. More
particularly, the apparatus is capable of sorting such particles
having different characteristics into different containers or
receptacles with a high degree of accuracy.
In recent years, accurate high speed machines have been devised for
measuring and indicating various characteristics of small particles
such as biological cells. One such machine is described and
claimed, for instance, in U.S. Pat. No. 3,705,771 dated Dec. 12,
1972 for a PHOTOANALYSIS APPARATUS, and assigned to the same
assignee as the present application. However, there is a continuing
important need for a machine which will very accurately and very
rapidly sort such particles into two or more groups having
different characteristics. This sorting function is particularly
needed for purposes of medical diagnosis, and for medical research.
There are many different particle characteristics which can be the
basis for the sorting or segregation. The sorting is particularly
valuable in instances where the particles or cells having the
unique characteristics are present in a very small proportion to
the total, making it difficult to obtain information about the
unique particles without physically separating those particles from
the main body of particles in which they occur.
So far as is known to the present inventor, no reasonably
economical, practical, and reliable machine has been heretofore
generally available which offers particle sorting at a rate
comparable to the operating rate at which the machine described in
the above patent can measure and indicate different particle
characteristics.
Accordingly, it is an object of the present invention to provide an
improved particle sorter for very small particles such as
biological cells.
Another object of the invention is to provide an economical
particle sorter which operates very rapidly.
The present invention involves the discovery that very rapid and
very satisfactory particle sorting may be carried out by employing
the technology referred to as fluidics, and particularly by
employing digital fluidic amplifiers which are normally thought of
as logic circuit devices to be incorporated into fluidic computing
or control systems which are analogous to comparable electrical
control systems. Surprisingly, it has been discovered in accordance
with the present invention that the principles of digital fluidic
amplifiers or switches can be very efficiently adapted in the
design of a particle sorter for microscopic particles.
Another object of the invention is to provide an improved particle
sorter incorporating at least one digital fluidic amplifier, or
digital switch, as a basic element.
Further objects and advantages of the invention will be apparent
from the following description and the accompanying drawings.
In carrying out the invention there is provided apparatus for
sorting small particles such as biological cells while the
particles are suspended in a liquid comprising a housing defining a
detection chamber, means for moving the particle suspending liquid
through said housing in a stream to convey the particles through
the stream. A detection means is associated with said detection
chamber for detecting differences in particle characteristics and
operable to provide electrical signals which vary in accordance
with said differences in particle characteristics. A digital
fluidic amplifier is provided having an inlet connected to the
outlet of said chamber to receive the particle stream, said fluidic
amplifier having a switching chamber communicating with said inlet
and at least two different outlet ports communicating with said
switching chamber, with an electrical transducer coupled to receive
electrical signals from said detection means and operable to
provide control signals to said fluidic amplifier. Said fluidic
amplifier is operable in response to said control signals to switch
the liquid particle carrying stream entering the inlet thereof from
a first outlet port to a second selected outlet port in response to
a predetermined particle characteristic control signal from said
transducer.
In the accompanying drawings:
FIG. 1 is a schematic side view, partly in section, illustrating a
preferred embodiment of the invention.
FIG. 2 is a representation corresponding to FIG. 1, but showing an
alternative embodiment of the invention.
FIG. 3 is a simplified schematic diagram illustrating a
modification of the invention employing a plurality of fluidic
amplifiers.
FIG. 4 is a simplified schematic diagram illustrating a
modification of the invention employing a plurality of combinations
of detection chambers and digital fluidic amplifiers connected in
tandem.
Referring more particularly to FIG. 1, there is shown an optical
chamber formed by a glass tube member 10 which is clamped by means
not shown to a digital fluidic switch chamber housing member 14,
the two members being sealed together by a liquid-tight annular
seal 18. The liquid 19 containing the particles to be observed
enters the apparatus through a tube 20 centrally disposed within
the funnel-shaped entrance portion 24 of the cylindrical central
bore 26 of the member 10. Another liquid 23 also enters the mouth
24 of the central bore 26 and forms a sheath of liquid for the
liquid 19 containing the particles.
The velocity and volume of flow of the particle-bearing liquid 19
and the sheath liquid 23 entering the mouth 24 of the central bore
26 are such as to cause the stream of particle-bearing liquid to be
narrowed down, as shown at 28, into a very narrow stream 29 having
a maximum dimension of the same order of magnitude as the maximum
dimension of the particles being carried by the stream. For
instance, this dimension may be in the order of 25 microns. The
particles of greatest interest are often somewhat smaller than
this, being in the range from 1 to 10 microns in diameter. The
funnel-shaped entrance portion 24 of the cylindrical member 10 is
preferably provided with an exponential function shape to provide
for smooth non-turbulent flow of the liquids at the critical
position 28 where the particle-carrying liquid is narrowed down.
Typically, the particle-carrying liquid may be an aqueous solution
and the sheath liquid 23 may be water.
The stream 29 of particles is illuminated by a beam of light
emitted by a light source 30 which preferably consists of a laser
together with an appropriate system of lenses as described more
fully in the above-mentioned patent. One satisfactory type of
laser, for instance, is a helium-neon laser. The laser and the
associated lenses provide a very narrow beam in which the pattern
of the illumination of the beam at point 38 where it strikes the
particles is preferably a very narrow ellipse which appears to be a
thin line of light transverse to the stream of particles.
Electrical photoresponsive pick-up elements are arranged around the
outside of cylindrical chamber member 10 to detect different
optical reactions of each particle to illumination from the beam.
These elements are illustrated at 40, 42, 44, 47, and 48 which are
all connected to provide signals to an apparatus 46, and elements
50 and 52 connected to provide signals to apparatus 54. The
apparatus 54 may be combined with the apparatus of 46, but is
separately shown to simplify the drawing. The apparatus 46 may
include amplifiers, logic circuitry, digital counters, and
electronic display devices. The circuits within the apparatus 46
may be carried out in accordance with the teachings of a prior U.S.
Pat. No. 3,662,176 issued May 9, 1972 on an invention of Louis A.
Kamentsky and Isaac Klinger for A PHOTO-OPTICAL PARTICLE ANALYSIS
METHOD AND APPARATUS which is assigned to the same assignee as the
present application. The apparatus 46 is sometimes referred to
hereinafter as "circuits 46."
When a unique particle characteristic is detected which signifies
the presence of a unique particle which is to be segregated from
the other particles, the detection circuits 46 send out an
electrical signal through a delay circuit 49, a gate circuit 51,
and connections 56 to an electrical transducer 58 which is mounted
within the fluidic switch housing 14. This causes a switching of
the particle-carrying stream from one outlet port to another.
The fluidic amplifier housing 14 defines a switching chamber 60
having a first outlet port 62, and a second outlet port 64. The
flow of liquid through the optical chamber 10 is in a laminar
(non-turbulent) mode. In the absence of a signal at the transducer
58, the stream of fluid emanating from the bore 26 of the chamber
10 continues to flow in a laminar mode into the inlet 66 of the
fluidic amplifier housing 14, and continues in the laminar mode
through the switching chamber 60, and out through the first outlet
port 62, which is in direct axial alignment with the inlet 66.
However, when the transducer 58 is energized, it induces turbulence
into the liquid stream. In the presence of turbulence, the stream
tends to attach itself to the nearest side wall 68. This "wall
attachment" effect causes the stream to follow the side wall 68 to
the second outlet port 64.
The wall attachment effect is sometimes referred to as the "Coanda
effect" in honor of the Rumanian engineer, Henry Coanda, who
discovered it. This effect is well recognized in the literature as
the basis for many fluidic digital switching devices. The wall
attachment effect is caused by a "bubble" of low pressure adjacent
to the exit of the nozzle formed by the inlet 66 at the beginning
of the near side wall 68. This low pressure bubble causes the
stream of liquid to bend towards the wall 68 and to become stable
in this deflected course of travel. Thus, as long as the stream
remains turbulent, by reason of the application of a signal to the
transducer 58, the stream remains deflected to the second outlet
port 64. However, upon the discontinuance of the signal to
transducer 58, laminar flow is reestablished, the wall attachment
effect dissappears, and the stream returns to the first outlet port
62.
The speed of operation of the circuits 46 in providing a signal to
the transducer 58 is properly correlated with the speed of the
liquid stream of particles and the dimension between the optical
detection point 38 and the transducer 58 to provide for switching
of the desired particles from port 62 to port 64. The dynamic
response in the speed of operation of the fluidic switch is also a
factor. For this purpose, the delay circuit 49 is provided between
the circuits 46 and transducer 58. The delay circuit 49 may be
adjustable to properly correlate the timing of the signal to
transducer 58 with the arrival of the particle to be switched by
the transducer. One of the important advantages of this preferred
embodiment of the invention is that the response of the system is
extremely rapid since the transmission of signals is accomplished
entirely electrically to the very point in the inlet 66 of the
fluidic switch where turbulence must be induced. This is in
contrast to other fluidic switches in which a signal is transmitted
through a column of fluid to the control point.
The transducer 58 is preferably a piezoelectric crystal which is
capable of physical deformation in response to an applied voltage.
The excitation signal applied to the crystal 58 is preferably an
alternating current at a frequency corresponding to the natural
resonant frequency of the crystal 58 and its surrounding fluid so
as to provide maximum mechanical output in response to the
available electrical input energy. Thus, the individual signals
applied to the transducer 58 are usually in the form of bursts of
alternating current gated by gate 51 from an oscillator 53. The
resultant alternating mechanical changes in the piezoelectric
crystal 58 are very efficient in inducing turbulence in the liquid
stream so as to assure immediate initiation of the wall attachment
effect.
While the transducer is shown as embedded in a side wall of the
inlet nozzle 66 of the fluidic amplifier on the same side as the
wall 68, it will be understood that the transducer may be effective
also in other locations communicating with the inlet 66, or in the
switching chamber 60 in close proximity to inlet 66. The
piezoelectric crystal is preferred as the transducer for this
purpose. However, other types of transducers, such as
electromagnetically energized acoustic vibration transducers may be
employed.
The optical chamber member 10 is a glass tube having a cylindrical
inner bore 26. However, the interior cavities of the digital
fluidic amplifier defined by the housing 14 may be circular or
non-circular in cross-section, or essentially two-dimensional in
nature. Thus, the inlet 66 may include a transition from a circular
shape to a rectangular shape, or the inlet 66 may simply be
rectangular in shape throughout its length, having a minimum
cross-sectional dimension which is at least as great as the
diameter of the bore 26 of the optical chamber 10. The transition
of the stream of liquid from the circular cross-section bore 26 to
the rectangular inlet 66 of the fluidic amplifier does not disturb
the flow sufficiently to create tubulence. Thus, the stream retains
its laminar-characteristic until it is made turbulent by the
transducer 58. The interior cavities of the housing 14 can be
rounded at the corners, and the inlet 66 and the outlet ports 62
and 64 can actually be circular in cross-section without
interfering with the operation of the device as described
above.
The ports 62 and 64 are connected to suitable collectors or
containers, not shown. Either, or both, of the separate collected
cell bearing liquids can be run through the apparatus again to
accomplish an additional refinement in the sorting operation by
again sorting for the same characteristic, or for still another
characteristic. After a run is completed, the apparatus may be
flushed with a saline solution or water. To assure complete
collection of all of the unique cells sorted at port 64, flushing
liquid may be applied at the inlet 23 while outlet port 62 is
blocked so that all of the flushing liquid is necessarily directed
through the outlet port 64, carrying any sorted particles which
remained in the port 64 and the associated passage to the collector
or container associated with that port. The apparatus is then
preferably completely flushed to prevent contamination of a new
sample with the remains of a completed sample.
A number of different characteristics of the particles may be
optically detected in the chamber 10 and used as a basis for the
sorting of the particles. For instance, the electrical
photoresponsive pick-up element 40 is arranged in direct line with
the beam to measure the degree of extinction of illumination by
each particle. In the absence of a particle at the intersection of
the beam, or in the absence of any substantial extinction, the beam
strikes the element 40 without any substantial diminution.
As illustrated in the drawing, the beam diverges to a certain
extent after having been converged at the center of the chamber at
38. The effective divergence in a practical embodiment has been
limited to approximately one degree on each side of the center line
of the beam as measured from the particle scanning point 38 at the
center of the chamber. Thus, photoresponsive pick-up elements 42
and 44 are arranged on opposite sides of the direct beam and can be
used to measure illumination scattered out of the direct beam by
the particles over a selected range of angles from one degree up to
a predetermined angular limit. For instance, this range of angles
may be from one degree to nine degrees. As shown in the drawing,
the photoresponsive pick-up elements 42 and 44 may be electrically
connected in parallel so that electrical signals resulting from
illumination scattered on either side of the beam and detected by
elements 42 and 44 will be registered at the electrical apparatus
46. Additional pairs of photoresponsive pick-up elements for
detecting scattered light at other ranges of angles may be provided
as shown at 47 and 48. For instance, this additional pair of
pick-up elements may detect scatter over the scatter angle range
from 9 degrees to 22 degrees.
Scattering of illumination from the particles in the reverse
direction, called "back scattering" can also be detected by
photoresponsive elements 50 and 52 arranged on the same side of the
chamber as the light source 30 and connected in parallel to an
electrical pick-up and recording apparatus 54. Apparatus 54 may be
combined with the apparatus 46, but it is shown separately here to
simplify the drawing.
The portion of the apparatus for detecting different particle
characteristics as just described above may preferably be carried
out in accordance with the teachings of the patent first mentioned
above. In addition to detecting different particle characteristics
by extinction and by scatter, distinctive particle characteristics
may also be determined on the basis of fluorescent radiation
reactions from the particles to illumination of the particles as
taught in that prior patent. When fluorescent radiation reactions
are desired, an argon ion laser may be used as the source of
illumination. Furthermore, sophisticated combinations of particle
measurement characteristics may be employed for controlling the
particle sorting operation. For instance, electrical summations or
differences of different signals may be employed at selected
threshold values for determining when a particle should be
segregated from the mainstream. Such circuits are described in the
aforementioned U.S. Pat. No. 3,662,176.
The present invention may preferably be implemented and carried out
by combining and incorporating features of an invention described
and claimed in a companion Pat. application Ser. No. (Docket 1305)
entitled PARTICLE SORTER WITH SEGREGATION INDICATOR filed
concurrently with the present application by Louis A. Kamentsky and
Isaac Klinger and assigned to the same assignee as the present
application. The following features disclosed in FIG. 1 relate to
the subject matter of that concurrently filed application:
A source of illumination such as a small incandescent lamp is
provided, as indicated at 55, which directs a beam through a
central slit in an optical mask 57 and thus through the particle
carrying stream to an optical pick-up element, or photocell, 59.
The optical pick-up element 59 is connected to an amplifier 61
which may be connected at 63 to control the variable delay circuit
49. The illumination from the lamp 55 traversing the particle
stream to the photoelectric pick-up element 59 is effective to
detect the passage of particles in the particle stream. Since this
combination including pick-up 59 is located near the transducer 58,
it is in a position to measure the arrival of the particle upon
which the transducer 58 is intended to be effective, and to thus
measure the travel time of the particle from the initial detection
point 38 to the transducer 58.
While the pick-up element 59 is not located at the identical
position of the transducer 58, the travel time of particles from
point 38 to pick-up element 59 is proportional to the travel time
from point 38 to the transducer 58 so that the interval until the
arrival of particles at pick-up element 59 is a measure of the
travel time to the transducer 58. Thus, the signals from pick-up
element 59 amplified by the amplifier 61 may be used to control the
delay circuit 49 to provide an exact match of the operation of the
electrical circuits energizing transducer 58 with the velocity of
the particle-carrying liquid. This provides an important
enhancement in the precision and accuracy of operation of the
sorter.
In accordance with another feature, another light source in the
form of an incandescent lamp 65 provides a beam of light through a
slit in an optical mask 67 which crosses the passage for port 64 to
a photoelectric pick-up element 69 which is connected to an
amplifier 71. By means of this apparatus associated with the
photoelectric pick-up 69, the passage of a particle through the
port 64 can be detected so as to positively indicate that the
sorting operation has been successfully accomplished with respect
to that particle. A very useful means for indicating this success
is a cathode ray oscilloscope 73 which is connected through a
switch 75 to receive the amplifier sort indication signal from the
amplifier 71, preferably as a vertical deflection on the
oscilloscope.
In order to provide a correlated horizontal sweep, the scope 73 is
preferably connected through a connection 77 to receive from the
circuits 46 an indication of the passage of the particle at the
original detection point 38, the signal on 77 being used to trigger
the initiaion of the horizontal sweep on the scope 73. Thus, the
vertical deflection caused by the signal from amplifier 71 will
occur at a predictable horizontal position on the scope 73 which is
related to the velocity of the particle-carrying liquid through the
apparatus.
Alternatively, a similar detection of the passage of particles to
the port 62 may be accomplished by the combination of a lamp 79, an
apertured mask 81, a photoresponsive pick-up element 83, and an
amplifier 85 connected thereto. The switch 75 is arranged to selet
the output from amplifier 71 or from amplifier 85 for indication on
the oscilloscope 73.
The pick-up 83 for port 62 may be used to indicate that the
particles passing through the port 62 have not been selected to be
segregated into port 64. For instance, the apparatus may be checked
for proper sorting operation by introducing a sample in which all
particles are of a class which should be segregated into the port
64. With such a sample, whenever the sorting operation is begun,
vertical deflections on the oscilloscope 73 derived from the
pick-up element 83 for port 62 should disappear completely.
The signal from the pick-up element 83 may also be employed in
place of the signal from pick-up element 59 for controlling the
delay circuit 49. Assuming that the main body of particles in the
initial liquid stream continues on through the discharge port 62,
the time interval of travel of an individual particle from the
initial detection point 38 to the vicinity of the pick-up element
83 will be a predictable function of the interval for the travel of
an individual particle from point 38 to the transducer 58.
Accordingly, the measurement available from pick-up element 83 is
an appropriate signal for controlling the delay circuit 49 in order
to properly time the gating of the signals to the transducer 58.
The pick-up element 59 and the associated apparatus may be omitted
from the system if the signal from pick-up element 83 is used to
control the delay circuit 49.
It is not absolutely necessary, in order to obtain the advantages
of these features to have a direct connection, such as connection
63, from one of the pick-up amplifiers to control the delay circuit
49. In other words, an open loop system may be employed in which
the delay interval is measured, such as by the indication on the
oscilloscope 73, and that measurement is then used to manually set
the delay on the variable delay circuit 49. However, the direct
connection 63 from the amplifier 61 to the delay circuit 49
provides the advantage of continuous and automatic adjustment of
the delay to compensate for any fluctuation in the velocity of the
particle-carrying stream.
In accomplishing the purposes of the pick-ups 69 and 83 in
indicating the accomplishment of the sort function, it may also be
useful to have counters attached for actuation from the amplifiers
71 and 85 for indicating and storing a registration of the numbers
of the particles which have been sorted into the respective ports
62 and 64. It is contemplated that the circuits 46 may include
counters for counting the total number of particles and also for
counting particles having unique characteristics to be detected and
upon which the sort is to be based. Accordingly, the counters
attached to the amplifiers 71 and 85 which indicate the numbers of
particles sorted into the two channels can be compared with the
counts registered by the counters within the apparatus 46 to
accurately determine the efficiency of the sorting operation.
It will be understood that the pick-up elements 59, 69, and 83, and
the associated apparatus, are shown schematically in order to
simplify and clarify the drawing. In a preferred embodiment of the
invention, each of these combinations of apparatus is preferably
rotated ninety degrees about the axis of the associated liquid
channel so that the direction of the light beam in each instance is
directly perpendicular to the plane of the section shown in the
drawing. Thus, in such a preferred arrangement, the pick-up element
59, for instance, would appear on the wall of the inlet 66 directly
behind the liquid stream. In such a preferred physical embodiment,
the lamp 55 and the pick-up 59 can be arranged and positioned
exactly at the axial position of the transducer 58 within the inlet
66 so as to provide an exact measurement of the interval until the
arrival of a particular particle at the transducer 58.
The switching chamber 60, and the ports 62, 64, and 66 may
preferably be formed by providing the chamber housing 14 in two
parts, one part containing cavities to provide the chamber and
ports, and the other part constituting a cover which is attached
over the other part to enclose the cavities. Thus, the part of the
housing 14 containing the cavities may contain the pick-up elements
59, 69, and 83 in the back walls thereof, and the light sources 55,
65, and 79, together with their associated optical masks, may be
attached to, or be a part of, the cover member. These positions
may, of course, be reversed.
Other modifications are also possible. For instance, fiber optics
may be employed to carry the illumination from a lamp, such as 55,
to the channel, such as 66, where it is needed. Instead, or in
addition, fiber optics may be used to convey the light beam from
the channel 66 to the photoelectric pick-up 59. The use of fiber
optics may be particularly advantageous because of the space
limitations in the vicinity of the channels being monitored, the
optical fibers requiring much less space than the lamp and
photoelectric pick-up elements. Also, light sources other than
incandescent lamps may be employed.
In order to simplify the drawings and descriptions of the
modifications of the invention illustrated in FIGS. 2, 3, and 4,
the pick-up elements 59, 69, and 83, and the associated apparatus,
are not illustrated or described in connection with these other
figures. However, it will be understood that the principles of the
features associated with these pick-up elements are equally
applicable to these modifications. For instance, in FIG. 2, three
outlet ports are provided from the switching chamber and it will be
understood that photoelectric sort detectors, such as elements 69
and 83, may be employed for each of those three outlet ports.
FIG. 2 illustrates an alternative embodiment of the invention in
which the switching chamber 60 is modified as shown at 60A, in a
modified fluidic amplifier housing 14A, to provide an additional
outlet port 70 and added control ports 72 and 74. A control fluid
under pressure may be supplied to the switching chamber 60A through
the control ports 72 and 74 through the respective supply lines 76
and 78 under the control of electromagnetically operated valves 80
and 82. These valves may be controlled from the apparatus 46
through the electrical connections respectively indicated at 84 and
86.
As illustrated in the drawing, the opposite walls 68 and 69 of the
switching chamber 60A are essentially equidistant from the nozzle
formed by the inlet 66. Accordingly, they are equally capable of
displaying the wall attachment efect in the presence of turbulence
in the liquid stream, as previously described above. Thus, just as
the wall attachment effect to wall 68 directs the liquid stream to
outlet port 64, the wall attachment effect at wall 69 leads the
fluid stream to the outlet port 70. Once established at one wall,
the wall attachment effect will be maintained and will hold the
stream to one of these two walls, and thus direct the stream to one
of these two outlet ports, as long as turbulence continues. In
order to determine which direction the stream will take, a signal
must be supplied from one of the control ports 72 or 74. If the
signal takes the form of a supply of control fluid under pressure,
it has the effect of shifting the stream away from the wall
adjacent to the control port and over to the opposite wall. Thus,
if control fluid is supplied to the switching chamber 68 through
the control port 74, the supply of fluid destroys the low pressure
bubble at the wall 69, causing the stream to shift to the opposite
wall 68. A similar, but opposite effect is available if a vacuum is
applied at the control port 74. This reduces the pressure at the
wall 69, tending to increase the wall attachment at the wall 69 and
to switch the device so that the liquid stream is pulled away from
the wall 68 and to the wall 69 and to the port 70.
The embodiment of FIG. 2 is capable of several different
alternative modes of operation. For instance, if the switching
chamber 60A is fabricated in an asymmetrical form such that the
wall 68 is substantially closer to the nozzle formed by the inlet
66 than the wall 69, then without any control signals at the
control ports 72 and 74, the switching action will be identical to
that described above for the embodiment of FIG. 1. Thus, if a
signal is supplied to the transducer 58, the resulting turbulence
will cause the stream to attach to the wall 68 and to be directed
to port 64. In the absence of a signal at transducer 58, the
laminar flow will continue, and the stream will be directed through
the outlet port 62.
However, with the asymmetrical configuration, if a fluid pressure
control signal is applied at the control port 72 while the
turbulent condition exists by reason of the operation of the
transducer 58, then the wall attachment effect at wall 68 will be
discontinued, and the stream will attach to wall 69, even though
that wall is more distant from the nozzle formed by inlet 66. Thus,
in such an arrangement, with an asymmetrical configuration, the
second control port 74 is not necessary to provide a three way
switching effect to the three outlet ports 62, 64, and 70.
Summarizing the operation of such an asymmetrical form of the
invention: with neither the transducer 58 nor the control port 72
being effective, laminar flow continues, and the stream exits
through the center port 62. With only the turbulence transducer 58
energized, the resultant turbulence causes the stream to attach to
wall 68 and to discharge through port 64. With both the turbulence
transducer 58 and the control port 72 in operation, the stream is
directed to the far wall 69 and to the outlet port 70. Since the
wall attachment effect is self-sustaining in the presence of
continued turbulence, the signal from control port 72 need not be
continued to maintain the discharge at port 70.
In the symmetrical form actually illustrated in FIG. 2, a signal
must be supplied from one of the control ports substantially
concurrently with the signal to the transducer 58 in order to make
it certain that the stream is directed to the correct outlet port.
When no cells are selected to be segregated from the stream, no
signals are supplied to the transducer 58 or to the valves 80 and
82. In these circumstances, the flow remains laminar, and the
liquid is discharged through the center port 62. However, upon the
detection of a cell or particle of a first type to be segregated,
signals are supplied substantially concurrently to the transducer
58 and to open the valve 82, respectively causing the stream to
become turbulent, and providing a control port signal at 74 causing
the stream to be switched to the wall 68 and the out-let port 64.
Upon the detection of a particle of a second category to be
segregated, signals are supplied to the transducer 58 to initiate
or maintain turbulence, and to open valve 80 to the control port
72, causing the stream to switch to the wall 69 and the outlet port
70. Thus, in this embodiment, there is the capability of sorting
the particles into three categories in a single pass through the
apparatus.
In another alternative mode of operation, the transducer 58 remains
on at all times, the stream is always turbulent, and switching of
the stream is always exclusively controlled by the control ports 72
and 74 so that the stream is always emitted either from the port 64
or the port 70. The center port 62 is not used. Also, with the
versatility provided by the modification of FIG. 2, there is
presented the possibility of achieving a faster response of the
fluidic amplifier in terminating the turbulent flow to one of the
non-axial outlet ports, such as 64, when the transducer is turned
off, by concurrently providing a short signal from the associated
control port 72. This sharply terminates the "wall attachment" at
wall 68 while laminar flow is being re-established.
In the broader sense, the valves 80 and 82 may also be referred to
as electrical transducers. They perform the function of transducers
in that they convert electrical signals to fluidic control signals
which cause the switching of the liquid stream from one outlet port
to another. The switching fluid used at the ports 72 and 74 should
be compatible with the liquid in the main stream, but it need not
be identical to that liquid. Thus, where the liquid in the main
stream is water, the switching fluid may be water, or air, or some
other compatible fluid.
For the sake of simplicity in the drawing, the valves 80 and 82 are
shown only schematically. However, it will be understood that they
may be electromagnetically operated valves, or they may operate on
some other principle. For instance, they may be piezoelectric
crystal actuated valves in which crystal movement in response to a
voltage serves to open the valve. While not so illustrated, the
valves 80 and 82 are preferably built into the housing 14A, as
close as possible to the exits of the associated control ports 72
and 74. By this means, the transit time of signals initiated
through the valves is substantially reduced since the signals are
transmitted substantially the entire distance to the point of use
by electrical means rather than by means of a column of fluid.
It is possible to modify the embodiment of FIG. 2 to provide for at
least two more side ports corresponding to ports 64 and 70 by
arranging such ports in a circular pattern around the central port
62. Thus, another side port (not shown) can be arranged behind the
central port 62, and another one can be arranged in front of the
central port 62. Such an arrangement requires the addition of
appropriate valves and control ports corresponding to the valves 80
and 82 and the control ports 72 and 74.
While not illustrated in the drawings of either of the FIGS. 1 or
2, the side outlet ports 64 and 70 may be provided with vent
passages to reduce the pressure in the outlet ports to enhance the
wall attachment effect and to reduce any possible impairment of the
switching action which might be caused by a build-up of pressure in
the receptacles to which the respective outlet ports are
attached.
The embodiment of FIG. 2 provides greater versatility than the
embodiment of FIG. 1, but the FIG. 1 embodiment is definitely
preferred. One of the major advantages is that the control ports 72
and 74 are not required, and the problem of purging the switching
chamber when different samples are to be used, and when cleaning is
required, is greatly simplified by the elimination of these control
ports.
In the embodiments of both FIGS. 1 and 2, the optical detection
chamber 10 is preferably composed of glass, and the switching
chamber housing 14 or 14A may preferably be composed of a
transparent synthetic resin material. While illustrated as separate
housings which can be disassembled from one another, it is obvious
that these housings can be combined in a unitary structure.
As illustrated in the simplified schematic diagram of FIG. 3,
additional fluidic amplifier switches may be employed downstream
from the fluidic amplifier switch of the system of FIG. 1, or in
that of FIG. 2. This provides the possibility of still further
sorting and segregation of particles on a single pass through the
apparatus. The downstream fluidic amplifiers may also be connected
to receive control signals from circuits 46.
FIG. 3, at the left side, illustrates in simplified schematic form
an apparatus corresponding to the apparatus of FIG. 1, with
corresponding parts correspondingly numbered. The main stream
outlet port 62 is connected to a second fluidic amplifier switch
housing 114 which may correspond exactly in structure and operation
with the fluidic switch contained within the housing 14 and
described above in connection with FIG. 1. The fluidic switch 114
responds to a signal from circuits 46 on a connection indicated at
146 through a delay circuit 149 and a gate circuit 151 to control a
signal from an oscillator 153 to a transducer within the switch
housing 114. By this means, particles which were not sorted and
segregated in the first fluidic amplifier within housing 14, and
diverted to the outlet port 64, continue on through the port
connection 62, through the switch 114, and may be sorted and
segregated according to a second particle characteristic into an
outlet port 164. If the particles are not so segregated, they
continue on in the main stream outlet port 162 from the housing
114.
Thus, FIG. 3 illustrates the principle that a plurality of fluidic
amplifier switches may be controlled to accomplish several sorting
operations based on signals obtained from a single optical chamber
10. In the operation of the system of FIG. 3, it will be understood
that the delay interval provided by the delay circuit 149 is
appropriately lengthened to accommodate for the additional interval
required for the particles to travel from the chamber 10 to the
digital fluidic amplifier 114.
FIG. 4 is a simplified schematic diagram illustrating the principle
that two sets of the apparatus shown and described in connection
with FIG. 1 above can be connected in tandem to accomplish
successive detection and sorting operations by two independent
optical detections of particle characteristics accompanied by
associated sorting operations. Thus, in FIG. 4, the combination of
apparatus at the left side of the figure corresponds precisely with
the apparatus illustrated and described above in connection with
FIG. 1. The apparatus on the right of the figure again duplicates
the apparatus shown and described above in connection with FIG. 1,
all of the parts being numbered with corresponding part numbers,
but with a prefix digit 2 added to each number. Thus, FIG. 4
illustrates the principle that a plurality of the apparatus
combinations of FIG. 1 may be connected in tandem to accomplish
successive sorts on the same sample.
While the larger combinations illustrated in FIG. 3 and FIG. 4 are
shown and described as being related to the apparatus of FIG. 1, it
will be quite obvious that the principle of the use of multiple
digital fluidic amplifiers and multiple combinations of sorting
apparatus connected in tandem may be applied as well to the
apparatus of FIG. 2.
The invention has been described above entirely in terms of a
liquid particle carrying stream. While a liquid handling apparatus
is preferred, it will be understood that the principles of the
invention can be employed with a gaseous fluid particle carrying
stream using a gas such as air.
While this invention has been shown and described in connection
with particular preferred embodiments, various alterations and
modifications will occur to those skilled in the art. Accordingly,
the following claims are intended to define the valid scope of this
invention over the prior art, and to cover all changes and
modifications falling within the true spirit and valid scope of
this invention.
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