CA2298300A1 - Method and apparatus for sorting and separating particles from a fluid suspension - Google Patents
Method and apparatus for sorting and separating particles from a fluid suspension Download PDFInfo
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Abstract
A method and apparatus for sorting and separating particles from a fluid suspension using gas bubbles. The fluid suspension containing the particles of interest is passed through a channel and interrogated to determine the presence of a particle. If a particle is detected, the fluid is subjected to energy to cause the formation of the bubble downstream of the interrogation point. The formation of the bubble is synchronized with the arrival of the particle and the particle is displaced by the bubble into another channel for collection or further processing.
The energy for forming the bubble is supplied by an external laser. The device lends itself to manufacture as a disposable unit and is suitable for sorting and separating cells from a biological specimen.
The energy for forming the bubble is supplied by an external laser. The device lends itself to manufacture as a disposable unit and is suitable for sorting and separating cells from a biological specimen.
Description
TITLE: METHOD AND APPARATUS FOR SORTING AND SEPARATING
PARTICLES FROM A FLUID SUSPENSION
FIELD OF THE INVENTION
The present invention relates to cytological specimen testing and more particularly to a method and apparatus for sorting and separating biological cells or material using the motive force of gas bubbles.
BACKGROUND OF THE INVENTION
In the fields of medicine and biology, the detection of rare or unusual cells within cytological specimens is a common activity. Two techniques are commonly utilized for cell detection. The first technique involves examination of a slide through a microscope and is classified as a static detection method. The second technique is considered dynamic and involves the use of a flow cytometer.
The static detection technique of rare cellular events forms the basis of a wide range of cancer screening tests, including the well-known Pap test for cervical cancer precursors. The static detection method involves arranging the specimen on the surface of a microscope slide and then visually examining the specimen under high magnification through the microscope. The static detection technique has the advantage of allowing repeated examinations of the specimen before a final decision is made. This improves the intrinsic accuracy of the test when performed by a skilled and trained technician.
However, as the cells of interest, i.e. the rare or unusual cells will be mixed in with perhaps hundreds of thousands of unimportant cells, there is a strong possibility that the presence of the unimportant cells will interfere with the accurate detection of the cells of interest.
The known dynamic detection methods fall into the general category of flow cytometry. Flow cytometry is a known technique for making rapid measurements on particles or cells as they flow in a fluid stream past a sensing point. By making individual cell measurements, rare events (i.e. unusual cells or cell formations) can be extracted from the background (i.e. cytological specimen) in a way that is not possible using bulk measurement in the static detection technique. However, in most cases the flow cytometry technique only provides a single pass of the fluid stream flow and as a result there is only one opportunity to detect the cells of interest or the target cells.
In the art, there remains a need for a system which combines the accuracy of the static detection technique with the rapid measurement time of the dynamic flow cytometry technique.
BRIEF SUMMARY OF THE INVENTION
The present invention provides method and apparatus which utilizes gas bubbles for sorting and separating particles from a fluid suspension. The invention is suitable for sorting and separating cells in a fluid containing a biological specimen.
The cell sorting and separation system utilizes a dynamic flow cytometric process to identify target cells (i.e. cells of interest) in the cytological specimen which is carried in a fluid stream. At an appropriate point, the cells of interest are directed from the primary fluid stream into a collection chamber, a secondary sorting path or into a secondary fluid stream. The diverted cells provide an enriched specimen which may then be prepared in a slide format and examined using static rare event detection techniques.
According to another aspect of the invention, the cells of interest or target cells in the primary fluid stream are diverted by applying a gas bubble impulse at the appropriate point in the flow, so that the diverted portion of the cellular fluid stream contains a higher concentration of the target cells. The gas bubble for diverting the target cells is generated by a high-intensity solid state laser.
In a first aspect, the present invention provides a device for selecting and separating particles of interest from a fluid suspension, the device comprises: (a) an input port for receiving the fluid suspension; (b) a main channel, the input port being formed at one end of the main channel, and the main channel having an output port at the other end, the main channel defining a flow path for the fluid; (c) a bubble forming chamber, the bubble forming chamber being connected to the main channel and being coupled to an energy source for receiving energy to form a bubble within fluid contained in the bubble forming chamber; (d) an interrogation port, the interrogation port being located between the input port and the bubble forming chamber; and (e) a diversion channel, the diversion channel being located across from the bubble forming chamber and providing a conduit for particles displaced by the bubble formed in the bubble forming chamber.
PARTICLES FROM A FLUID SUSPENSION
FIELD OF THE INVENTION
The present invention relates to cytological specimen testing and more particularly to a method and apparatus for sorting and separating biological cells or material using the motive force of gas bubbles.
BACKGROUND OF THE INVENTION
In the fields of medicine and biology, the detection of rare or unusual cells within cytological specimens is a common activity. Two techniques are commonly utilized for cell detection. The first technique involves examination of a slide through a microscope and is classified as a static detection method. The second technique is considered dynamic and involves the use of a flow cytometer.
The static detection technique of rare cellular events forms the basis of a wide range of cancer screening tests, including the well-known Pap test for cervical cancer precursors. The static detection method involves arranging the specimen on the surface of a microscope slide and then visually examining the specimen under high magnification through the microscope. The static detection technique has the advantage of allowing repeated examinations of the specimen before a final decision is made. This improves the intrinsic accuracy of the test when performed by a skilled and trained technician.
However, as the cells of interest, i.e. the rare or unusual cells will be mixed in with perhaps hundreds of thousands of unimportant cells, there is a strong possibility that the presence of the unimportant cells will interfere with the accurate detection of the cells of interest.
The known dynamic detection methods fall into the general category of flow cytometry. Flow cytometry is a known technique for making rapid measurements on particles or cells as they flow in a fluid stream past a sensing point. By making individual cell measurements, rare events (i.e. unusual cells or cell formations) can be extracted from the background (i.e. cytological specimen) in a way that is not possible using bulk measurement in the static detection technique. However, in most cases the flow cytometry technique only provides a single pass of the fluid stream flow and as a result there is only one opportunity to detect the cells of interest or the target cells.
In the art, there remains a need for a system which combines the accuracy of the static detection technique with the rapid measurement time of the dynamic flow cytometry technique.
BRIEF SUMMARY OF THE INVENTION
The present invention provides method and apparatus which utilizes gas bubbles for sorting and separating particles from a fluid suspension. The invention is suitable for sorting and separating cells in a fluid containing a biological specimen.
The cell sorting and separation system utilizes a dynamic flow cytometric process to identify target cells (i.e. cells of interest) in the cytological specimen which is carried in a fluid stream. At an appropriate point, the cells of interest are directed from the primary fluid stream into a collection chamber, a secondary sorting path or into a secondary fluid stream. The diverted cells provide an enriched specimen which may then be prepared in a slide format and examined using static rare event detection techniques.
According to another aspect of the invention, the cells of interest or target cells in the primary fluid stream are diverted by applying a gas bubble impulse at the appropriate point in the flow, so that the diverted portion of the cellular fluid stream contains a higher concentration of the target cells. The gas bubble for diverting the target cells is generated by a high-intensity solid state laser.
In a first aspect, the present invention provides a device for selecting and separating particles of interest from a fluid suspension, the device comprises: (a) an input port for receiving the fluid suspension; (b) a main channel, the input port being formed at one end of the main channel, and the main channel having an output port at the other end, the main channel defining a flow path for the fluid; (c) a bubble forming chamber, the bubble forming chamber being connected to the main channel and being coupled to an energy source for receiving energy to form a bubble within fluid contained in the bubble forming chamber; (d) an interrogation port, the interrogation port being located between the input port and the bubble forming chamber; and (e) a diversion channel, the diversion channel being located across from the bubble forming chamber and providing a conduit for particles displaced by the bubble formed in the bubble forming chamber.
In a second aspect, the present invention provides a method for separating target particles from a fluid suspension containing one or more of said particles, the method comprises the steps of: (a) passing the fluid suspension containing the target particles past an interrogation point and determining the presence of one or more of the target particles in the fluid suspension; (b) applying an energy source to heat a portion of the fluid suspension proximate said target particles determined in step (a) to form a bubble in the fluid suspension;(c) utilizing said formed bubble to divert the target particles for further processing.
BRIEF DESCRIPTION OF THE DRAWINGS
Reference will now be made to the accompanying drawings, which show by way of example, a preferred embodiment of the present invention, and in which:
Fig. 1 is a diagrammatic view of a cell sorting and separation apparatus according to the present invention;
Fig. 2 is a schematic view of the main components of the cell sorting and separation apparatus of Fig. 1;
Fig. 3 is a schematic plan view of the main components of the cell sorting and separation apparatus of Fig. 1;
Figs. 4(a) to 4(f) schematically depict the operation of the cell sorting and separation apparatus according to the present invention;
BRIEF DESCRIPTION OF THE DRAWINGS
Reference will now be made to the accompanying drawings, which show by way of example, a preferred embodiment of the present invention, and in which:
Fig. 1 is a diagrammatic view of a cell sorting and separation apparatus according to the present invention;
Fig. 2 is a schematic view of the main components of the cell sorting and separation apparatus of Fig. 1;
Fig. 3 is a schematic plan view of the main components of the cell sorting and separation apparatus of Fig. 1;
Figs. 4(a) to 4(f) schematically depict the operation of the cell sorting and separation apparatus according to the present invention;
Fig. 5 shows a flow chart from the method steps for sorting and separating cells from a fluid stream according to the present invention;
Fig. 6 shows in schematic form a cascaded configuration for the cell sorting and separation apparatus according to the present invention;
Fig. 7 shows in schematic form a serial configuration for the cell sorting and separation apparatus according to the present invention; and Fig. 8 shows in schematic form a parallel configuration for the cell sorting and separation apparatus according to the present invention.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
Reference is first made to Fig. 1 which shows a cell sorting and separation device according to the present invention and indicated generally by reference 10. As will be described in more detail below, the cell sorting and separation device 10 together with an external laser 11 forms a system for effectively and rapidly separating cells of interest (i.e. target cells) from a cellular or biological specimen, indicated generally by reference S, carried in a fluid stream which is passed through the device 10. In the preferred embodiment, the cell sorting and separation device 10 comprises a cartridge with no moving parts and the cell sorting and separation mechanism is actuated by the external laser 11. Advantageously, the cartridge for the cell sorting and separation device 10 lends itself to being disposable and can be manufactured relatively cheaply in large quantities.
Fig. 6 shows in schematic form a cascaded configuration for the cell sorting and separation apparatus according to the present invention;
Fig. 7 shows in schematic form a serial configuration for the cell sorting and separation apparatus according to the present invention; and Fig. 8 shows in schematic form a parallel configuration for the cell sorting and separation apparatus according to the present invention.
DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENT
Reference is first made to Fig. 1 which shows a cell sorting and separation device according to the present invention and indicated generally by reference 10. As will be described in more detail below, the cell sorting and separation device 10 together with an external laser 11 forms a system for effectively and rapidly separating cells of interest (i.e. target cells) from a cellular or biological specimen, indicated generally by reference S, carried in a fluid stream which is passed through the device 10. In the preferred embodiment, the cell sorting and separation device 10 comprises a cartridge with no moving parts and the cell sorting and separation mechanism is actuated by the external laser 11. Advantageously, the cartridge for the cell sorting and separation device 10 lends itself to being disposable and can be manufactured relatively cheaply in large quantities.
Preferably, the laser 11 is situated exterior to the cell sorting and separation apparatus. This arrangement provides a non-contactive system in which the laser 11 is isolated from possible contamination and the cell sorting and separating device 10 lends itself to a disposable device which makes the system suitable for medical applications. Furthermore, as the cell sorting and separating device 10 is externally actuated and with minimal moving parts, the device 10 is well-suited to mass manufacturing which further covers the per unit cost of the disposable device. Other high energy output devices may be suitably substituted for the laser 11.
As shown in Fig. 1 (and also in Figs. 2 and 3 in which like references indicate like elements), the cell sorting and separation device 10 comprises a cartridge 20.
The cartridge 20 is designed to be a disposable device with no moving parts and also lends itself to mass manufacturing processes such as plastic injection molding. On the cartridge 20 is formed a main channel 21, a bubble chamber 22, a diversion channel 24 and an interrogation port 26.
The main channel 21 provides the primary flow path for a cellular specimen S suspended in fluid. The main channel 21 has an input port 28 and an output port 30. The input port 28 couples the main channel 21 to a supply vessel (not shown) containing the cellular specimen fluid and receives a portion of the cellular specimen fluid S. Once the cellular specimen fluid S passes through the device 10, it exists at the output port 30 which is connected to a waste collection vessel (not shown). While the device 10 is described in the context of a biological or cellular specimen separating application, it will be understood that the invention has wider applicability to other applications _ 7 _ where selected particles are to be separated from a fluid suspension.
The main channel 21 first takes the cellular specimen fluid S past the interrogation port 26. The interrogation port 26 provides a first actuation point in the device 10 and involves using a sensing mechanism (indicated generally by reference 27) to identify particles or cells of interest (i.e. target cells) in the cellular specimen fluid S. The interrogation port 26 utilizes optical, electromagnetic or other sensing mechanisms to identify the target cells in the fluid specimen S. The sensing mechanism 27 at the interrogation port 26 may comprise a laser system which measures forward and right-angle light scattering in the cellular fluid specimen S.
Or, the sensing mechanism 27 may comprise a fluorescence system utilizing an ultraviolet light source and a visible light detector to measure absorption or scattering of the ultraviolet light. Or, the sensing mechanism 27 may comprise some other type of non-contactive measurement that can differentiate between or among the particles in the fluid S flowing in the main channel 21. The sensing mechanism 27 at the interrogation port 26 produces an output when the cells of interest are detected at the interrogation port 26. The interrogation output is utilized for the second actuation point in the device 10.
The second actuation point in the device 10 occurs at the bubble chamber 22. The interrogation output is registered by a control system 12 and used to activate the laser 11 in a timed sequence. When the target cells or particles have moved to the bubble chamber 22, the control system 12 activates the external laser 11 which directs a high energy light beam into the bubble chamber 22. The _ g _ control system 12 may comprise a hardware (e. g. logic) circuit, a microcontroller or microprocessor suitably operating under program (i.e. firmware) control, or a computer system (e.g. personal computer) operating on a known platform (e.g. Windows T'" or UNIX) .
The bubble chamber 22 includes a window 23 for emitting the high energy light beam. The energy from the light beam is absorbed by the cellular specimen fluid S in the chamber 22 and the resultant heating in the fluid S
causes a bubble B to form. As the bubble B forms and grows in the blind bubble chamber 22 the target particles or cells are pushed into the diversion channel 24 and directed into a processing chamber 27. The bubble chamber 22 may include an optically absorptive pad 25 to facilitate the heating (i.e. formation of the bubble B) in cellular fluid specimen S which are not sufficiently absorptive to ensure rapid formation of the bubble B.
The processing chamber 27 may simply comprise a collection port. Or, the processing chamber 27 may include a port 29 leading to a secondary cell sorting and separation device as described below. Or, the diversion channel 24 may comprise a short connection to a second channel which runs parallel to the main channel 21 as described in more detail according to another embodiment of the invention.
Reference is next made to Figs. 4(a) to 4(f) and the flow chart in Fig. 5, which both illustrate the operation of the cell sorting and separation device 10 according to the present invention. The first step in Fig.
4(a) involves setting up the flow of cellular fluid specimen S in the main channel 21 in order to carry the g _ specimen past the interrogation port 26 (block 101 in Fig.
5). The second step in Fig. 4(b) involves sensing or detecting a particle P (or cell) of interest as it is carried past the interrogation port 26 (block 102 in Fig.
5). The presence of the particle P is registered by the control system 12 and used to activate the external laser 11. As shown in Fig. 4(c), after a short interval of time the target particle P is in position in the main channel 21 next to the bubble chamber 22 (block 104 in Fig. 5). The next step in Fig. 4(d) involves activating the external laser 11 to apply a short pulse of energy to the bubble chamber 22 (block 106 in Fig. 5). The pulse of laser energy locally heats the fluid S and induces the creation of a bubble B in the fluid specimen S sitting in the bubble chamber 22. As the bubble B evolves and grows in size, the bubble B acts to push a portion of the fluid S in the main channel 21 into the diversion channel 24 (block 108 in Fig.
5). Provided the formation of the bubble B is synchronized to the passage of the target particle P past the bubble chamber 22, the target particle P is re-directed into the diversion channel 24. The particle P moves through the diversion channel 24 to the processing chamber 27 where it is stored or moved to another stage for further processing (block 110 in Fig. 5).
Reference is next made to Fig. 6 which shows a cascaded arrangement of the cell sorting and separation apparatus according to another aspect of the present invention. The cascaded arrangement is indicated generally by reference 200 and comprises a first cell sorting and separation device 210a and a second cell sorting and separation device 210b. The devices 210a, 210b are as described above for device 10 and include respective main channels 221a, 221b, bubble chambers 222a, 222b, diversion channels 224a, 224b, interrogation ports 226a, 226b, input ports 228a, 228b, and output ports 230a, 230b. An external laser 211a and 211b is provided for each of the bubble chambers 222a and 222b respectively. Alternatively, a single laser 211 may be utilized for both devices 210a, 210b with an appropriate beam control mechanism.
As shown in Fig. 6, instead of a processing chamber 227a, the diversion channel 224a for the first device 210a is coupled to the input port 228b of the second device 210b. The cascaded arrangement 200 allows the selection of target cells or particles to be gradually refined without comprising the rate of specimen processing.
The first cell sorting and separation device 210a provides the primary cell sorting and separation operation and is operated at a high speed because the target cells are a very small fraction of the total cellular specimen. The high flow rates achievable in the main channel 221a of the primary device 210a minimize the processing time. While the increased processing time is desirable, the selection of target cells in the cellular suspension may not be as accurate as required, and target cells together with other non-specific cells may be driven into the diversion channel 224a. The second cell sorting and separation device 210b provides a secondary sorting and separation operation to further refine the specimen received from the primary diversion channel 224a. Since the absolute quantity of cellular specimen for sorting and separation is reduced considerably, the flow rate in the main channel 221b of the second cell sorting and separation device 210b can be significantly reduced. This, in turn, improves the selectivity of the bubble sorting and separation mechanism and results in an enriched cellular specimen sample at the output 230b of the second cell sorting and separation device 210b.
It will be appreciated that any number of cell sorting and separation devices 210 may be cascaded together as described above with reference to Fig. 6. The number of devices 210 cascaded together will depend on the complexity of the sorting and separation procedure to be performed.
Reference is next made to Fig. 7 which shows a serial arrangement of the cell sorting and separation apparatus according to another aspect of the present invention. The serial arrangement is indicated generally by reference 300 and comprises three cell sorting and separation devices 310a, 310b, and 310c connected in a serial arrangement as shown in Fig. 7. The individual devices 310a, 310b, 310c are similar to the device 10 described above and include respective main channels 321a, 231b and 321c, bubble chambers 322a, 322b, 322c, diversion channels 324a, 324b, 324c, interrogation ports 326a, 326b, 326c, input ports 328a, 328b, 328c and output ports 330a, 330b, 330c.
As shown in Fig. 7, the input port 328a for the first device 310a provides the input port for the serial arrangement 300. The output port 330a of the first device 310a is connected to the input port 328b of the second device 310b. Similarly, the output port 330b of the second device 310b is connected to the input port 328c of the third device 310c. The output port 330c of the third device 310c provides the output for the serial device 300.
The serial arrangement 300 of Fig. 7 results in a device having a main channel 321 with multiple interrogation ports 326a, 326b and 326c, multiple bubble chambers 322a, 322b and 322c, and multiple diversion channels324a, 324b and 324c. As the cellular suspension flows through the main channel 321, the outputs generated from the respective interrogation points 326a, 326b, 326c permit target cells or particles to be diverted into the appropriate diversion channel 324a, 324b and 324c. This is done by the control system issuing the appropriate signal sequence to induce the bubbles in the respective bubble chambers 322a, 322b and 322c.
Reference is next made to Fig. 8 which shows a parallel configured cell sorting and separation device 400 according to another aspect of the invention. The parallel cell sorting and separation device comprises a main channel 421 and a secondary channel 431. The main channel 421 includes a bubble chamber 422, a diversion channel 424, an interrogation port 426, an input port 428 and an output port 430. The secondary channel 431 is coupled to the main channel 421 through the diversion channel 424.
For the parallel configured device 400, the flow in the main channel 421 runs parallel to the flow in the secondary channel 431. At low fluid flow velocities, e.g.
low Reynolds numbers, there will be little fluid exchange between the main channel 421 and the secondary channel 431, apart from diffusion. When a target cell is detected at the interrogation port 426, the external laser is activated to form a bubble in the bubble forming chamber 422. The laser-induced bubble pushes the target cell out of the main channel 421 and into the secondary channel 431. From the secondary channel 431, the target cell is directed to further processing steps.
The cell sorting and separation device 10 according to the present invention has the advantage of providing a fast reaction time since the gas bubble can be created very rapidly and applied to the fluid stream. The fast reaction times also translates into smaller effective geometries for device 10 which may be manufactured as a disposable.
In summary, the cell sorting and separation device 10 according to the present invention embodies the following features. The sorting and separation mechanism is externally actuated which allows the device 10 to be designed as a simple and inexpensive disposable cartridge.
The device 10 includes no moving parts that may foul or otherwise contaminate the fluids. The device 10 is not subject to wear. The device 10 can be operated at high-speed. The device 10 is self-regenerating and re-setting.
The sorting and separation mechanism requires little space and may be located in a range of positions in the cartridge.
The present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. For instance, the device is suitable for sorting and separating particles other than just cells, from a fluid suspension. Therefore, the presently discussed embodiments are considered to be illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
As shown in Fig. 1 (and also in Figs. 2 and 3 in which like references indicate like elements), the cell sorting and separation device 10 comprises a cartridge 20.
The cartridge 20 is designed to be a disposable device with no moving parts and also lends itself to mass manufacturing processes such as plastic injection molding. On the cartridge 20 is formed a main channel 21, a bubble chamber 22, a diversion channel 24 and an interrogation port 26.
The main channel 21 provides the primary flow path for a cellular specimen S suspended in fluid. The main channel 21 has an input port 28 and an output port 30. The input port 28 couples the main channel 21 to a supply vessel (not shown) containing the cellular specimen fluid and receives a portion of the cellular specimen fluid S. Once the cellular specimen fluid S passes through the device 10, it exists at the output port 30 which is connected to a waste collection vessel (not shown). While the device 10 is described in the context of a biological or cellular specimen separating application, it will be understood that the invention has wider applicability to other applications _ 7 _ where selected particles are to be separated from a fluid suspension.
The main channel 21 first takes the cellular specimen fluid S past the interrogation port 26. The interrogation port 26 provides a first actuation point in the device 10 and involves using a sensing mechanism (indicated generally by reference 27) to identify particles or cells of interest (i.e. target cells) in the cellular specimen fluid S. The interrogation port 26 utilizes optical, electromagnetic or other sensing mechanisms to identify the target cells in the fluid specimen S. The sensing mechanism 27 at the interrogation port 26 may comprise a laser system which measures forward and right-angle light scattering in the cellular fluid specimen S.
Or, the sensing mechanism 27 may comprise a fluorescence system utilizing an ultraviolet light source and a visible light detector to measure absorption or scattering of the ultraviolet light. Or, the sensing mechanism 27 may comprise some other type of non-contactive measurement that can differentiate between or among the particles in the fluid S flowing in the main channel 21. The sensing mechanism 27 at the interrogation port 26 produces an output when the cells of interest are detected at the interrogation port 26. The interrogation output is utilized for the second actuation point in the device 10.
The second actuation point in the device 10 occurs at the bubble chamber 22. The interrogation output is registered by a control system 12 and used to activate the laser 11 in a timed sequence. When the target cells or particles have moved to the bubble chamber 22, the control system 12 activates the external laser 11 which directs a high energy light beam into the bubble chamber 22. The _ g _ control system 12 may comprise a hardware (e. g. logic) circuit, a microcontroller or microprocessor suitably operating under program (i.e. firmware) control, or a computer system (e.g. personal computer) operating on a known platform (e.g. Windows T'" or UNIX) .
The bubble chamber 22 includes a window 23 for emitting the high energy light beam. The energy from the light beam is absorbed by the cellular specimen fluid S in the chamber 22 and the resultant heating in the fluid S
causes a bubble B to form. As the bubble B forms and grows in the blind bubble chamber 22 the target particles or cells are pushed into the diversion channel 24 and directed into a processing chamber 27. The bubble chamber 22 may include an optically absorptive pad 25 to facilitate the heating (i.e. formation of the bubble B) in cellular fluid specimen S which are not sufficiently absorptive to ensure rapid formation of the bubble B.
The processing chamber 27 may simply comprise a collection port. Or, the processing chamber 27 may include a port 29 leading to a secondary cell sorting and separation device as described below. Or, the diversion channel 24 may comprise a short connection to a second channel which runs parallel to the main channel 21 as described in more detail according to another embodiment of the invention.
Reference is next made to Figs. 4(a) to 4(f) and the flow chart in Fig. 5, which both illustrate the operation of the cell sorting and separation device 10 according to the present invention. The first step in Fig.
4(a) involves setting up the flow of cellular fluid specimen S in the main channel 21 in order to carry the g _ specimen past the interrogation port 26 (block 101 in Fig.
5). The second step in Fig. 4(b) involves sensing or detecting a particle P (or cell) of interest as it is carried past the interrogation port 26 (block 102 in Fig.
5). The presence of the particle P is registered by the control system 12 and used to activate the external laser 11. As shown in Fig. 4(c), after a short interval of time the target particle P is in position in the main channel 21 next to the bubble chamber 22 (block 104 in Fig. 5). The next step in Fig. 4(d) involves activating the external laser 11 to apply a short pulse of energy to the bubble chamber 22 (block 106 in Fig. 5). The pulse of laser energy locally heats the fluid S and induces the creation of a bubble B in the fluid specimen S sitting in the bubble chamber 22. As the bubble B evolves and grows in size, the bubble B acts to push a portion of the fluid S in the main channel 21 into the diversion channel 24 (block 108 in Fig.
5). Provided the formation of the bubble B is synchronized to the passage of the target particle P past the bubble chamber 22, the target particle P is re-directed into the diversion channel 24. The particle P moves through the diversion channel 24 to the processing chamber 27 where it is stored or moved to another stage for further processing (block 110 in Fig. 5).
Reference is next made to Fig. 6 which shows a cascaded arrangement of the cell sorting and separation apparatus according to another aspect of the present invention. The cascaded arrangement is indicated generally by reference 200 and comprises a first cell sorting and separation device 210a and a second cell sorting and separation device 210b. The devices 210a, 210b are as described above for device 10 and include respective main channels 221a, 221b, bubble chambers 222a, 222b, diversion channels 224a, 224b, interrogation ports 226a, 226b, input ports 228a, 228b, and output ports 230a, 230b. An external laser 211a and 211b is provided for each of the bubble chambers 222a and 222b respectively. Alternatively, a single laser 211 may be utilized for both devices 210a, 210b with an appropriate beam control mechanism.
As shown in Fig. 6, instead of a processing chamber 227a, the diversion channel 224a for the first device 210a is coupled to the input port 228b of the second device 210b. The cascaded arrangement 200 allows the selection of target cells or particles to be gradually refined without comprising the rate of specimen processing.
The first cell sorting and separation device 210a provides the primary cell sorting and separation operation and is operated at a high speed because the target cells are a very small fraction of the total cellular specimen. The high flow rates achievable in the main channel 221a of the primary device 210a minimize the processing time. While the increased processing time is desirable, the selection of target cells in the cellular suspension may not be as accurate as required, and target cells together with other non-specific cells may be driven into the diversion channel 224a. The second cell sorting and separation device 210b provides a secondary sorting and separation operation to further refine the specimen received from the primary diversion channel 224a. Since the absolute quantity of cellular specimen for sorting and separation is reduced considerably, the flow rate in the main channel 221b of the second cell sorting and separation device 210b can be significantly reduced. This, in turn, improves the selectivity of the bubble sorting and separation mechanism and results in an enriched cellular specimen sample at the output 230b of the second cell sorting and separation device 210b.
It will be appreciated that any number of cell sorting and separation devices 210 may be cascaded together as described above with reference to Fig. 6. The number of devices 210 cascaded together will depend on the complexity of the sorting and separation procedure to be performed.
Reference is next made to Fig. 7 which shows a serial arrangement of the cell sorting and separation apparatus according to another aspect of the present invention. The serial arrangement is indicated generally by reference 300 and comprises three cell sorting and separation devices 310a, 310b, and 310c connected in a serial arrangement as shown in Fig. 7. The individual devices 310a, 310b, 310c are similar to the device 10 described above and include respective main channels 321a, 231b and 321c, bubble chambers 322a, 322b, 322c, diversion channels 324a, 324b, 324c, interrogation ports 326a, 326b, 326c, input ports 328a, 328b, 328c and output ports 330a, 330b, 330c.
As shown in Fig. 7, the input port 328a for the first device 310a provides the input port for the serial arrangement 300. The output port 330a of the first device 310a is connected to the input port 328b of the second device 310b. Similarly, the output port 330b of the second device 310b is connected to the input port 328c of the third device 310c. The output port 330c of the third device 310c provides the output for the serial device 300.
The serial arrangement 300 of Fig. 7 results in a device having a main channel 321 with multiple interrogation ports 326a, 326b and 326c, multiple bubble chambers 322a, 322b and 322c, and multiple diversion channels324a, 324b and 324c. As the cellular suspension flows through the main channel 321, the outputs generated from the respective interrogation points 326a, 326b, 326c permit target cells or particles to be diverted into the appropriate diversion channel 324a, 324b and 324c. This is done by the control system issuing the appropriate signal sequence to induce the bubbles in the respective bubble chambers 322a, 322b and 322c.
Reference is next made to Fig. 8 which shows a parallel configured cell sorting and separation device 400 according to another aspect of the invention. The parallel cell sorting and separation device comprises a main channel 421 and a secondary channel 431. The main channel 421 includes a bubble chamber 422, a diversion channel 424, an interrogation port 426, an input port 428 and an output port 430. The secondary channel 431 is coupled to the main channel 421 through the diversion channel 424.
For the parallel configured device 400, the flow in the main channel 421 runs parallel to the flow in the secondary channel 431. At low fluid flow velocities, e.g.
low Reynolds numbers, there will be little fluid exchange between the main channel 421 and the secondary channel 431, apart from diffusion. When a target cell is detected at the interrogation port 426, the external laser is activated to form a bubble in the bubble forming chamber 422. The laser-induced bubble pushes the target cell out of the main channel 421 and into the secondary channel 431. From the secondary channel 431, the target cell is directed to further processing steps.
The cell sorting and separation device 10 according to the present invention has the advantage of providing a fast reaction time since the gas bubble can be created very rapidly and applied to the fluid stream. The fast reaction times also translates into smaller effective geometries for device 10 which may be manufactured as a disposable.
In summary, the cell sorting and separation device 10 according to the present invention embodies the following features. The sorting and separation mechanism is externally actuated which allows the device 10 to be designed as a simple and inexpensive disposable cartridge.
The device 10 includes no moving parts that may foul or otherwise contaminate the fluids. The device 10 is not subject to wear. The device 10 can be operated at high-speed. The device 10 is self-regenerating and re-setting.
The sorting and separation mechanism requires little space and may be located in a range of positions in the cartridge.
The present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. For instance, the device is suitable for sorting and separating particles other than just cells, from a fluid suspension. Therefore, the presently discussed embodiments are considered to be illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Claims (16)
1. A device for selecting and separating particles of interest from a fluid suspension, said device comprising:
(a) an input port for receiving the fluid suspension;
(b) a main channel, said input port being formed at one end of said main channel, and said main channel having an output port at the other end, said main channel defining a flow path for the fluid;
(c) a bubble forming chamber, said bubble forming chamber being connected to said main channel and being coupled to an energy source for receiving energy to form a bubble within fluid contained in said bubble forming chamber;
(d) an interrogation port, said interrogation port being located between said input port and said bubble forming chamber; and (e) a diversion channel, said diversion channel being located across from said bubble forming chamber and providing a conduit for particles displaced by said bubble formed in said bubble forming chamber.
(a) an input port for receiving the fluid suspension;
(b) a main channel, said input port being formed at one end of said main channel, and said main channel having an output port at the other end, said main channel defining a flow path for the fluid;
(c) a bubble forming chamber, said bubble forming chamber being connected to said main channel and being coupled to an energy source for receiving energy to form a bubble within fluid contained in said bubble forming chamber;
(d) an interrogation port, said interrogation port being located between said input port and said bubble forming chamber; and (e) a diversion channel, said diversion channel being located across from said bubble forming chamber and providing a conduit for particles displaced by said bubble formed in said bubble forming chamber.
2. The device as claimed in claim 1, wherein said energy source comprises a high energy external laser.
3. The device as claimed in claim 1, wherein said interrogation port comprises an optical window, said optical window being coupled to an optical detector adapted for detection of the particle.
4. The device as claimed in claim 3, wherein said optical detector comprises a fluorescence system comprising a light source and a light detector.
5. The device as claimed in claim 2, wherein said bubble forming chamber includes a pad for absorbing energy from the laser and accelerating the formation of said bubble.
6. The device as claimed in claim 4, wherein said fluid suspension carries a cellular specimen and said particles comprise cells.
7. The device as claimed in claim 1, further including a second device for selecting and separating particles, said second device having an input port and said input port being coupled to the output of the diversion channel of said first device for receiving the target cells diverted from the main channel of the first device, and said second device having a main channel, a bubble forming chamber, an interrogation port and a diversion channel.
8. The device as claimed in claim 7, wherein said energy source comprises a high energy external laser.
9. The device as claimed in claim 8, wherein said bubble forming chamber for said first device includes a pad for absorbing energy from the laser and accelerating the formation of the bubble in the bubble chamber.
10. The device as claimed in claim 1, further including a second device for sorting and separating particles, said second device having an input port connected to the output port of the first device for receiving the flow of fluid suspension from the main channel of said first device, and said second device including a main channel, a bubble forming chamber, an interrogation port and a diversion channel.
11. The device as claimed in claim 2, wherein said device is manufactured as a disposable cartridge, said main channel being formed in the cartridge, said diversion channel being formed in the cartridge, and said bubble chamber being formed in the cartridge.
12. A device for selecting and separating particles of interest from a fluid suspension, said device comprising:
(a) an input port for receiving the fluid suspension;
(b) a main channel, said input port being formed at one end of said main channel, and said main channel having an output port at the other end, said main channel defining a flow path for the fluid;
(c) a bubble forming chamber, said bubble forming chamber being connected to said main channel and being coupled to an external laser for receiving energy to form a bubble within fluid adjacent said bubble forming chamber;
(d) an interrogation port, said interrogation port being located between said input port and said bubble forming chamber;
(e) a diversion channel, said diversion channel being located across from said bubble forming chamber and providing a conduit for particles displaced by said bubble formed in said bubble forming chamber; and (f) a secondary channel having an input port coupled to said diversion channel for receiving particles diverted from the fluid suspension in said main channel.
(a) an input port for receiving the fluid suspension;
(b) a main channel, said input port being formed at one end of said main channel, and said main channel having an output port at the other end, said main channel defining a flow path for the fluid;
(c) a bubble forming chamber, said bubble forming chamber being connected to said main channel and being coupled to an external laser for receiving energy to form a bubble within fluid adjacent said bubble forming chamber;
(d) an interrogation port, said interrogation port being located between said input port and said bubble forming chamber;
(e) a diversion channel, said diversion channel being located across from said bubble forming chamber and providing a conduit for particles displaced by said bubble formed in said bubble forming chamber; and (f) a secondary channel having an input port coupled to said diversion channel for receiving particles diverted from the fluid suspension in said main channel.
13. A method for separating target particles from a fluid suspension containing one or more of said particles, said method comprising the steps of:
(a) passing the fluid suspension containing the target particles past an interrogation point and determining the presence of one or more of said target particles in said fluid suspension;
(b) applying an energy source to the fluid to heat a portion of the fluid suspension proximate said target particles determined in step (a) to form a bubble in the fluid suspension;
(c) utilizing said formed bubble to divert said target particles for further processing.
(a) passing the fluid suspension containing the target particles past an interrogation point and determining the presence of one or more of said target particles in said fluid suspension;
(b) applying an energy source to the fluid to heat a portion of the fluid suspension proximate said target particles determined in step (a) to form a bubble in the fluid suspension;
(c) utilizing said formed bubble to divert said target particles for further processing.
14. The method as claimed in claim 13, wherein said step applying an energy source comprises activating a high energy laser.
15. The method as claimed in claim 13, wherein said step (c) of further processing comprises collecting said target particles for deposition on a slide.
16. The method as claimed in claim 14, wherein said step of further processing comprises repeating steps (a) to (b).
Applications Claiming Priority (2)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US11929299P | 1999-02-09 | 1999-02-09 | |
| US60/119,292 | 1999-02-09 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CA2298300A1 true CA2298300A1 (en) | 2000-08-09 |
Family
ID=31886291
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CA 2298300 Abandoned CA2298300A1 (en) | 1999-02-09 | 2000-02-08 | Method and apparatus for sorting and separating particles from a fluid suspension |
Country Status (1)
| Country | Link |
|---|---|
| CA (1) | CA2298300A1 (en) |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232468A (en) * | 2014-08-28 | 2014-12-24 | 中北大学 | Laser array coded and photoinduced cell separation device |
| CN104263644A (en) * | 2014-08-28 | 2015-01-07 | 中北大学 | Cell separation method based on laser array coding and photo-induction |
| CN113502207A (en) * | 2021-08-18 | 2021-10-15 | 长春长光辰英生物科学仪器有限公司 | Multifunctional cell sorting device based on laser system and operation method |
-
2000
- 2000-02-08 CA CA 2298300 patent/CA2298300A1/en not_active Abandoned
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104232468A (en) * | 2014-08-28 | 2014-12-24 | 中北大学 | Laser array coded and photoinduced cell separation device |
| CN104263644A (en) * | 2014-08-28 | 2015-01-07 | 中北大学 | Cell separation method based on laser array coding and photo-induction |
| CN104232468B (en) * | 2014-08-28 | 2016-03-30 | 中北大学 | Laser array coding and photoinduced cell separation apparatus |
| CN104263644B (en) * | 2014-08-28 | 2016-07-06 | 中北大学 | Laser array coding and photoinduced cell isolation method |
| CN113502207A (en) * | 2021-08-18 | 2021-10-15 | 长春长光辰英生物科学仪器有限公司 | Multifunctional cell sorting device based on laser system and operation method |
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