WO2007130923A2

WO2007130923A2 - Portable platform cytometric system with isolation chamber

Info

Publication number: WO2007130923A2
Application number: PCT/US2007/067831
Authority: WO
Inventors: Brenda Lee
Original assignee: Cellerant Therapeutics, Inc.
Priority date: 2006-05-01
Filing date: 2007-04-30
Publication date: 2007-11-15
Also published as: WO2007130923A3

Abstract

A cytometer for measuring particles includes a portable platform supporting an illumination table including a measuring site, a detection system for detecting at least one property of the particles in the measuring site, a fluidics system configured for transporting the particles from a sample source to the measuring site, the fluidics system including transfer tubing fluidly connecting the sample source to the measuring site, and an isolation chamber housing and environmentally sealing the illumination table and sample source from an ambient environment. A method of using the cytometer is also disclosed.

Description

PORTABLE PLATFORM CYTOMETRIC SYSTEM WITH ISOLATION CHAMBER

CROSS-REFERENCE TO RELATED APPLICATIONS

[oooi] This application claims benefit under 35 U. S. C.§ 119 to U. S. Provisional Application Serial No: 60/796,766 filed May 1, 2006, the disclosure of which is incorporated by reference herein in its entirety.

TECHNICAL FIELD OF THE INVENTION

[0002] This invention relates, in general, to a cytometric system and more particularly to a flow cytometer and methods for their use.

BACKGROUND OF THE INVENTION

[0003] In the fields of biochemistry, analytical chemistry, molecular biology, pathology, immunology, and similar fields, laboratory researchers increasingly use cytometry for the analysis, counting, measuring, and general examination of microscopic particles on a large scale. Flow cytometers in particular encapsulate or carry sample particles in a sheath fluid or flow cell in a measuring region.

[0004] Typically, flow cytometry involves labeling selected particles with fluorescent molecules (e.g. fluorochromes) that bind to the particles. A charge is then optionally applied to the particles. A nozzle supplies the particles one-by-one to a measuring region where they are illuminated. A detection system detects the reflection (or the emitted fluorescence from the excited fluorochrome dye) from the particles, which is then converted to an electrical charge that is amplified, measured, and analyzed. [0004] Modern flow cytometers are capable of measuring thousands of particles a second and a multitude of physical and chemical characteristics. Modern applications involve particle samples highly sensitive to an external environment. One limitation of modern flow cytometry involves the need for a sterile measuring environment and reduction of sample contact.

[0005] Prior art cytometric systems have employed a glove box or isolation box around the nozzle assembly and measuring region to minimize influence on the particles in the measuring region where environmental affects can skew data. However, these systems have failed to adequately maintain a sterile environment during the measuring and sorting process. Additionally, these systems have failed to adequately protect the users of such systems during sorting, cleaning of the glove box, and similar tasks.

[0006] An exemplar of the prior art is U.S. Patent No. 5,641,457 to Vardanega et al. which shows a cytometric system with an isolation chamber encapsulating a sterilization system including a measuring region, sorting region, cell station, and cooling station. Further, a flow region within the chamber is mechanically isolated from the isolation chamber so that movement of the chamber does not affect the measurement data.

[0007] While Vardanega discloses the particle sorter and collector disposed within the isolation chamber, the sample source and other selected components remain outside the isolation chamber. Thus, the Vardanega system makes cleaning cumbersome and less effective. In order to adequately clean the inside of the chamber and replace components such as the fluid tubing, the isolation chamber must be opened and interior components unfastened.

[0008] Furthermore, the Vardanega system does not adequately protect users because the sample source is located outside of the isolation chamber. Thus, the user may be exposed to sample, especially when cleaning the sample block, collar, and tubing. Samples trapped in the fluidic tubing can further fall out of the tubing during cleaning and expose users.

[0009] Additionally, the Vardanega system allows for the risk of skewed data from environmental influence because the Vardanega system does not isolate the detection system and emitted or reflected light from the measured particles, the detected data may be altered.

[ooio] Vardanega and other prior art systems also will not accommodate simply adding components to the isolation chamber. Modern cytometric systems include complex components for the delivery and measuring of sample at a measuring site. These components typically are paired with bulky equipment emitting vibrations, light, and other undesirable effects that can alter measurement data if placed within the chamber. Prior art isolation chambers are thus limited as to which components can be isolated. In the alternative, if such equipment is isolated, the isolation chamber would be large and unwieldy. Because "clean room" environments are highly costly and increase in cost with size, it is desirable to minimize their size.

[ooii] Prior art cytometric systems also have the additional limitation of being cumbersome and awkward to maintain. During cleaning, the entire isolation chamber needs to be opened, and the interior components must be removed and cleaned in a non-sterile, non-protective environment. Even when disassembled, the nooks and crannies inside the isolation chamber prevent adequate sterilization. Prior art systems further require the tedious task of passing cabling and tubing through the walls of the isolation chamber.

[ooi2] What is needed is a cytometric system that overcomes the above and other disadvantages. In particular, what is needed is a cytometric system isolating selected components to increase the accuracy of measurements and protect users from exposure to the sample without subjecting the measuring site to undesirable effects from system components. [000] Further, what is needed is a cytometric that is quick and easy to clean. Additionally, what is needed is a cytometric system where selected components are supported in a portable configuration.

BRIEF SUMMARY OF THE INVENTION

[ooi4] In summary, one aspect of the present invention is directed to a cytometric system for measuring particles. The system includes a portable platform supporting an illumination table including a measuring site, a detection system for detecting at least one property of the particles in the measuring site, a fluidics system configured for transporting the particles from a sample source to the measuring site, the fluidics system including transfer tubing fluidly connecting the sample source to the measuring site, and an isolation chamber housing and environmentally sealing the illumination table and sample source from an ambient environment.

[ooi5] In one embodiment, the portable platform may further support a particle sorting system configured for sorting the particles below the measuring site based upon the detection of the at least one property of the particles. The particle sorting system may include a particle sorting arm assembly including a sorting arm movably extending into the isolation chamber though a chamber slot, and an isolation boot sealingly received around the sorting arm and sealingly affixed to the isolation chamber.

[ooi6] The system may further include a cooling block for cooling particles collected after sorting. The cooling block is housed within the isolation chamber. The cytometric system may include a collection tube holder magnetically-mounted to the cooling block within the isolation chamber. The isolation chamber housing may be pressure-sealed.

[ooi7] In one embodiment, the isolation chamber is a glove box. The isolation chamber may include an interface panel in a side wall configured to interface between components in the isolation chamber and components external to the isolation chamber. The interface panel may be configured to receive banana plugs. [0018] The cytometric system may include a sample block for supporting the sample source, wherein the sample block is movably supported within the isolation chamber by a cam assembly to facilitate loading the sample source on the sample block. The transfer tubing of the system may include pinch valves mounted on the sample block to selectively control flow from the sample source to the measuring site.

[ooi9] The cytometric system may include a sample system including a sheath tank for providing sheath fluid to the measuring site, and including a waste tank for receiving waste fluid from the measuring site.

[0020] The illumination table may include at least one laser located outside of the isolation chamber and at least one illumination tube being configured to direct output from the at least one laser to the measuring site. A portion of the at least one illumination tube may be housed within the isolation chamber. The isolation chamber may include at least one environmentally- sealed port in one wall configured for accepting the at least one illumination tube therethrough. The illumination table may include a stage drive for horizontally aligning a sheath of particles in the measuring site with the laser output, wherein the stage drive includes a vertically extending knob for adjusting alignment.

[0021] The detection system may include at least one bank of detectors, a control unit, and a power source, wherein the at least one bank of detectors is housed within the isolation chamber.

[0022] The sample source and illumination table may be spaced from a bottom floor of the chamber to facilitate cleaning and decontamination of the isolation chamber.

[0023] The cytometric system maybe configured to detect and sort particles which are stem cells.

[0024] Another aspect of the present invention is directed to a cytometric system for measuring a sample of particles carried by a sheath fluid. The system includes an illumination table supporting a measuring site, an isolation chamber isolating an interior from the ambient environment, a detection system for detecting at least one property of the particles in the measuring site, the detection system including at least a bank of detectors, a camera having a collimating lens, a directing prism, and a power source, a fluidics system for transferring the sample to the measuring site, the fluidics system including at least a nozzle, sample source, and transfer tubing configured for transferring the sample from the sample source to the nozzle. The illumination table, sample source, supply tubing, and nozzle may be removably housed within the isolation chamber and environmentally isolated from an ambient environment.

[0025] In one embodiment, the cytometric system includes a particle sorting system configured for sorting the particles based upon the detection of at least one property of the particles in the sample. The at least one camera and at least one directing prism may be housed within the isolation chamber. The isolation chamber may be configured to allow removal of the fluidics system without disassembly of the isolation chamber. Preferably, the particles to be measured are living cells.

[0026] The cytometric system of the present invention has other features and advantages which will be apparent from or are set forth in more detail in the accompanying drawings, which are incorporated in and form a part of this specification, and the following Detailed Description of the Invention, which together serve to explain the principles of the present invention.

BRIEF DESCRIPTION OF THE DRAWINGS

[0027] FIG. 1 is a perspective view of a cytometric system utilizing a portable platform in accordance with the present invention.

[0028] FIG. 2 is a perspective schematic view of the portable platform of FIG. 1 within the cytometric system of FIG. 1.

[0029] FIG. 3 is an enlarged perspective view of portions of a fluidics system and sample system within the cytometric system of FIG. 1. [0030] FIG. 4 is an enlarged perspective view of a sample system illustrating a collar removed from a sample housing within the cytometric system of FIG. 1.

[0031] FIG. 5 is an enlarged perspective view of the sample system of FIG. 4 within the portable platform of FIG. 1 illustrating a lever in a lowered position facilitating access to a sample block of the sample system.

[0032] FIG. 6 is an enlarged perspective view of the of the sample system FIG. 4 within the portable platform of FIG. 1 illustrating the sample block in a removed position.

[0033] FIG. 7 is an enlarged perspective view of the top of the sample system of FIG. 4 within the portable platform of FIG. 1 shown with optional pinch valve and attached fluidics.

[0034] FIG. 8 is an enlarged side view of the sample system of FIG. 4 within the portable platform of FIG. 1 illustrating a pinch valve switch.

[0035] FIG. 9 is an enlarged perspective view of pinch valves on the interior of the portable platform of FIG. 1 within the cytometric system of FIG. 1.

[0036] FIG. 10 is an enlarged perspective view of an exterior control for the pinch valves of FIG. 9 on an interface panel of the portable platform of FIG. 1.

[0037] FIG. 11 is an enlarged perspective view of a stage drive and adjustment knobs of the illumination table within the portable platform of FIG. 1.

[0038] FIG. 12 is an enlarged front view of the cooling block of FIG. 1 within the portable platform of FIG. 1.

[0039] FIG. 13 is an enlarged front view of the cooling block of FIG. 12 illustrating the cooling block with a collection tube holder removed. [0040] FIG. 14 is an enlarged perspective view of a bottom portion of an illumination frame within the portable platform of FIG. 1.

[0041] FIG. 15 is an enlarged front view of the cytometric system of FIG. 1 shown with only the particle sorting arm and spacers.

[0042] FIG. 16 is an enlarged perspective view of an interface panel of the portable platform of FIG. 1.

[0043] FIG. 17 is an enlarged front view of the interface panel of FIG. 16 within the portable platform of FIG. 1 illustrating electrical wiring plugged into the panel.

[0044] FIG. 18 is an enlarged front view of the interface panel of FIG. 16 within the portable platform of FIG. 1 illustrating fluidic tubing plugged into the panel.

[0045] FIG. 19 is an enlarged perspective view of the backside of the portable platform of FIG. 1.

DETAILED DESCRIPTION OF THE INVENTION

[0046] Reference will now be made in detail to the preferred embodiments of the invention, examples of which are illustrated in the accompanying drawings. While the invention will be described in conjunction with the preferred embodiments, it will be understood that they are not intended to limit the invention to those embodiments. On the contrary, the invention is intended to cover alternatives, modifications and equivalents, which may be included within the spirit and scope of the invention as defined by the appended claims.

[0047] For convenience in explanation and accurate definition in the appended claims and detailed description, the terms "up" or "upper", "down" or "lower", "inside" and "outside" are used to describe features of the present invention with reference to the positions of such features as displayed in the figures. [0048] Turning now to the drawings, wherein like components are designated by like reference numerals throughout the various figures, attention is directed to FIG. 1, which illustrates a cytometric system, generally designated 30, in accordance with the present invention. The illustrated cytometric system is similar to the MOFLO cytometer currently sold by DakoCytomation of Denmark in conjunction with a workstation and electronics console. The MOFLO cytometer is described in the MOFLO Owner's Manual, Version 3.2, July 2002, the entire content of which is incorporated by reference herein.

[0049] While the illustrated system is used for flow cytometry on human blood cells, one will appreciate that the cytometric system can be used in myriad applications involving the multiparametric analysis of the physical or chemical properties of any number of microscopic particles and sorting of such particles using light detection. The methods of measuring sample particles and general configuration of cytometric systems are well known and will not be discussed in detail.

[ooso] In the illustrated embodiment in FIG. 1, a cytometric system 30 includes an electronics console 32, portable platform 33, and workstation 35. The electronics console includes a number of optional subsystems for the monitoring and analysis of data from components on the portable platform.

[0051] Turning to FIG. 2-6, portable platform 33 supports an illumination table 37, an illumination frame 39, and a detection system 40. The portable platform also supports a sample source 42 and at least a portion of a fluidics system generally designated 44. The illumination table supports a measuring site 46 through which sample particles pass for measuring in a known manner. Detection system 40 then detects at least one property of the particles as they pass through the measuring site. Such properties include, but are not limited to, particle volume, DNA, RNA, chromosome analysis, nuclear antigens, enzymatic activity, membrane fluidity, intracellular and surface antigens, pH, oxidative burst, and other physical and chemical properties. [0052] Turning to FIGs. 2-8, sample source 42 is housed within an isolation chamber 47. The sample source includes a sample housing 49. The sample source supplies the sample of particles to be measured at the measuring site. In the illustrated embodiment, a sample block 51 supports the sample source. Sample source 42 includes the components extending from the supply of sample to the sample block, but will generally be used interchangeably with sample assembly 53 which includes sample housing 49, sample block 51, and associated components unless otherwise noted.

[0053] The sample source includes a collar 54 and rotating cams 56. The cams lock the sample source into place thereby movably supporting the sample source within the isolation chamber. Thus, the sample may be loaded on the sample with greater ease. Although the sample source is depicted with a cam assembly, one will appreciate that other suitable configurations may be used to secure the sample source.

[0054] Fluidics system 44 includes a system of fluidic tubing for transporting fluids through cytometric system 30. Specifically, fluidics system 44 includes the fluidic connections to and from the sample source as well as a nozzle assembly 58. In the illustrated embodiment, the nozzle assembly is dimensioned and configured to provide a stream of single particles in a known manner, however, one will appreciate that other nozzle configurations may be utilized depending upon the application. The fluidics system also includes, in part, portions of sample source 42.

[0055] Transfer tubing 60 (see FIG. 7) transports particles from sample source 42 to nozzle assembly 58. The nozzle assembly separates the sample and delivers single particles to the measuring site. Supply tubing 61 connects the sample source to the supply of sample in the sample tank. Below the measuring site, the fluidics system includes additional tubing to transport waste fluid or sample to the waste tank.

[0056] Focusing on FIGs. 7-8, the transfer tubing extends through optional pinch valve 63 mounted on the sample source. The pinch valve allows a user to selectively control the flow from the sample source to the measuring site. The pinch valve also allows the flow to be shut off when disconnecting the sample block. More importantly, the pinch valve allows the transfer tubing to be readily removed from the supply system and avoids the need to clean valve chambers and/or valve assemblies as the pinch valves are never directly exposed to sample.

[0057] In the illustrated embodiment, the sample source is stainless steel and the transfer tubing is a non-reactive plastic. One will appreciate, however, that other suitable materials may be used for the sample source including, but not limited to, non-reactive metals, plastics, fibers, or other suitable materials. Suitable materials for the fluidics system tubing include, but are not limited to polyetheretherketone and other polyetherketones, polytetrafluoroethylene, and other suitable materials including non-reactive metals and fibers.

[0058] Focusing again on FIGs. 1-8, illumination table 37 supports the illumination and detection components within isolation chamber 47. The illumination table also includes an illumination source 65 located outside of the isolation chamber. In the illustrated embodiment, the illumination source is a laser; however, one will appreciate that other illumination apparatuses may be employed depending upon the application. The laser output is directed by at least one illumination tube 67 to the measuring site thereby illuminating particles for detection.

[0059] In one embodiment, the illumination tube is configured to extend the transmission length of the laser and direct the output of laser 65 to measuring site 46. A portion of the illumination tube near the measuring site is housed within isolation chamber 47, and a portion connected to the laser is housed outside the isolation chamber. The isolation chamber includes an environmentally-sealed port 68 in one wall configured for accepting the illumination tube therethrough. The sealed port is dimensioned to allow a tight fit of the illumination tube and includes a sealing ring or other sealing configurations. Although the illustrated embodiment is shown with one laser and corresponding illumination tube and sealed port, one will appreciate that the cytometric system may include more than one laser and illumination tube or other configurations not using any illumination tube depending on the application. [0060] At an end of the illumination tube, illumination table 37 supports measuring site 46. Turning to FIG. 11, an upper end of the illumination table is shown. A nozzle aperture 70 is configured to receive a lower end of nozzle assembly 58 therethrough. The illumination table includes a stage drive 72 for horizontally aligning particles in the measuring site with the laser output. In the illustrated embodiment, the stage drive includes a vertically-extending knob 74 for adjusting alignment of the measuring site. The knob allows a user to adjust the position of the illumination table relative to the laser output so that the particles in the measuring site will be aligned in the path of the laser output as desired. Knob 74 is optionally in a vertical orientation to allow a user to easily access the knobs above the illumination table and inside isolation chamber 47. One will appreciate that other suitable configurations for the stage drive and adjustment knob are envisioned including, but not limited to, use of a plurality of stage drives and controlling knobs.

[006i] Returning to FIGs. 1-2, a portion of detection system 40 is located in the vicinity of measuring site 46. The detection system generally includes a detection subsystem and electronics components such as a preamplifier, a control unit, and a power source. The detection subsystem generally includes components such as a collimating lens, pinhole apertures, pinhole camera, directing prisms, and a bank of detectors 75. In the illustrated embodiment, the bank of detectors, directing prisms, and pinhole camera are housed within the isolation chamber near the measuring site. However, any number of components may be chosen for isolation or placed outside the isolation chamber depending on the application specifications.

[0062] The components briefly described above are housed within or adjacent to the isolation chamber. Focusing now on the isolation chamber, the isolation chamber generally is configured for promoting a sealed clean room atmosphere. The isolation chamber generally allows for removal of interior components and removal of the isolation chamber itself from cytometric system 30 with minimal use of tools. In this regard, the interior components are affixed to the inner walls of chamber 47 such that chamber 47 is easier to clean and less likely to collect contaminant particles. The isolation chamber serves to isolate selected components discussed above from the ambient environment while promoting access to internal components and ease of use.

[0063] In the illustrated embodiment, the isolation chamber is a glovebox environmentally sealing an interior environment from an ambient environment. The chamber is pressure-sealed to allow a positive pressure within. In some aspects, the isolation chamber is similar to the glovebox systems sold by M.Braun, Inc. of Stratham, New Hampshire.

[0064] As shown in FIGs. 2 and 14-15, the sample source and illumination table are spaced from a bottom floor of isolation chamber 47 using spacers 77. The spacing facilitates cleaning and decontamination of the isolation chamber. Nooks and corners that tend to collect contaminants are minimized inside the isolation chamber by the use of the spacers. The spacing on floor of the isolation chamber also promotes the flow of air or gases in the chamber during evacuation and vaporous cleaning of the interior.

[0065] The spacers also serve a secondary purpose. The placement of illumination table 37 within isolation chamber 47 may result in an increased elevation relative to illumination tube 67 and laser 65. Thus, spacers 77 may also serve to vertically align the respective components for measuring.

[0066] The flow of the sample of particles can now be described in more detail. A sheath tank houses a sheath fluid on or adjacent the portable platform 33. Fluidics system 44 includes transfer tubing 60 configured for delivering sample from the sample source 42 to the measuring site. In the illustrated embodiment, the sample source is located adjacent to illumination frame 39 and illumination table 37, all of which reside within an isolation chamber 47.

[0067] Cytometric system 30 further includes a sheath tank for providing sheath fluid to the measuring site and a waste tank for receiving waste fluid. The sheath fluid is commonly used to carry the sample. At the nozzle exit, the particles to be measured

- I O - will be hydrodynamically focused in the core of the sheath stream. Alternative methods of delivering and carrying sample particles are envisioned depending on the application.

[0068] The fluidics system 44 includes supply tubing 61 for transferring the sample from the sample tank to the isolation chamber. In the illustrated embodiment, a section of the fluidics tubing external to the isolation chamber connects to a plug or interface panel 79 in a side wall. Another section of tubing then transfers the sample from the interface panel to the sample source.

[0069] In the illustrated embodiment, the fluidics tubing connects to the sample source via pinch valves 63. As depicted in FIGs. 7-10, the pinch valves facilitate removal of the transfer tubing and other components from the isolation chamber. Furthermore, the pinch valves obviate the need for decontamination and sterilization the valve interiors and/or valve chambers. On the interior of isolation chamber 47, a series of pinch valves control the flow of fluids through the isolation chamber walls and interface panel. As best seen in FIG. 10, a control knob 81 in the interface panel may optionally be provided to control the pinch valves from the exterior of the isolation chamber. In the illustrated embodiment, the control knob allows a user to select which pinch valves to actuate and subsequently which fluidic tubing to shut off.

[0070] Similarly, the flow of sample fluid to the sample source and sheath fluid to the nozzle assembly may be controlled with pinch valves. Although the use of pinch valves is preferred as shown in the illustrated embodiment, one will appreciate that other suitable means may be used to control the flow of fluid inside the isolation chamber provided that such means facilitate quick and easy removal of the fluidics tubing.

[007i] As shown in FIG. 8, the sample source optionally includes a support 82 on a top portion of the sample housing 49. The support is rigidly affixed to the sample housing at a bottom end. A top end of the support rigidly holds sample pinch valves 63. A section of the fluidics tubing running from the interface panel connects to the sample source at the pinch valves so that a user can twist the pinch valves closed. Similarly, the pinch valves adjacent to the interface panel can stop the flow from the sample tank.

[0072] Sample housing 49 further includes a cam assembly 84 which operably supports sample block 51 and the sample source 42 supported thereon to the sample housing. The cam assembly includes a lever 86 and cam member 88. With the lever in a down position, the cam member is lowered and the sample block may be readily removed (best seen in FIG. 6). With the sample block removed, the fluidics tubing is exposed for easy removal from the sample block. In the illustrated embodiment, the fluidics tubing is threadably engaged with the sample block, however other suitable quick-release means may be used. One will appreciate that other configurations may be used to releasably support the sample block within the sample housing.

[0073] Turning to FIGs. 16-18, interface panel 79 includes a bulkhead 89 configured to receive electrical wiring connectors. The bulkhead is secured to the walls of the isolation chamber. The interface panel is configured to interface between components inside and outside the isolation chamber such that the interior components can be connected to external components without exposing the isolation chamber interior to the ambient environment. For example, certain components on the interior of the isolation chamber may be connected to an external power source through the interface panel. In the illustrated embodiment, the interface panel is configured so that internal components may be connected to an inner face of the interface panel and the bulkhead outside the chamber may be connected to the outer part of the interface panel. As shown in FIGs 17-18, the interface panel may include connections for fluidic tubing and electrical wiring alike. In one embodiment, the interface panel includes adaptors for receiving banana plugs. Such configuration avoids the need for pass-through apertures in the wall of the isolation chamber and thus renders unnecessary o-rings and other similar seals which may fail under the caustic conditions of sterilization and decontamination. [0074] Returning to FIGs. 3 and 4, sample source 42 located within isolation chamber 47 includes collar 54 at a top end of the sample housing 49. The collar includes a flange designed to sit in a groove on top of the sample housing. The groove facilitates seating of the collar on the housing. Fluidics system 44 includes a stiff guiding portion 91. In the assembled position, the sample collar locates the guiding portion within the sample source. A user can then insert the guiding portion through the collar to connect to the sample block.

[0075] From sample source 42, fluidics system 44 transfers sample particles to the measuring site through nozzle assembly 58. As shown in FIG. 11, illumination frame 39 includes nozzle aperture 70. Above the measuring site, the particles to be measured are focused within a sheath of fluid. The nozzle assembly then separates the sample of particles into single particles for delivery through the aperture to the measuring site on the illumination table.

[0076] Turning to FIG. 19, in one embodiment, a rear wall of the isolation chamber separates the illumination table and detection components. A detection component extension member 93 is provided to space the external detection components from the measuring site so as to allow the wall of isolation chamber 47 to fit therebetween. In the illustrated embodiment, the power source, electrical components, and analytical components are placed outside the isolation chamber to obviate the need for increasing the size of the chamber. However, the components chosen for isolation within the isolation chamber and the manner for connecting them to external components may vary by application.

[0077] Once the measured particles pass through the measuring site, they may be sorted. Turning now to FIGs. 1-2 and 15, the portable platform is shown supporting an optional particle sorting system 95 configured for sorting the particles below the measuring site based upon the detection of at least one property of the particles. The particle sorting system optionally includes a particle sorting arm assembly 96 below illumination table 37. The sorting arm assembly includes a sorting arm 98 for deposition of sorted particles in multiwell plates or a matrix in a known manner. [0078] In accordance with the present invention, the sorting arm movably extends into the isolation chamber though a chamber slot 100. The slot is dimensioned such that it also allows the sorting arm to rotate about a pivot within or adjacent to a side wall of the isolation chamber and to otherwise provide clearance for the sorting arm to move in its usual an known manner. In the illustrated embodiment, the sorting arm includes an isolation boot 102 to prevent exposure of the interior of the isolation chamber to the ambient environment. The isolation boot is sealingly received around the sorting arm and sealingly affixed to the isolation chamber (see FIG. 15). In the illustrated embodiment, the sorting arm includes an annular groove to receive the boot in a sealed manner.

[0079] The insertion of sorting arm 98 through the rear wall of isolation chamber 47 with isolation boot 102 therebetween balances the need for access to the interior isolation chamber with the desirability of maintaining a sterile environment therein. Furthermore, the boot of the illustrated embodiment allows the sorting arm to be easily removed from the isolation chamber without disassembling the isolation chamber.

[0080] Turning to FIGs. 12-13, a cooling block 103 for cooling particles after sorting is shown. The cooling block is situated adjacent to the illumination table and within isolation chamber 47. A series of fluid tubing 105 circulates cold fluid through the cooling block. The cooling block may be located remotely from the measuring site and illumination frame. More than one cooling block or other cooling methods may also be used depending on the application.

[008i] In the illustrated embodiment, the cooling block includes a collection tube holder 107. In contrast to the existing art, the collection tube holder mounts to the cooling block using magnetics. In this manner, the collection tube may be easily removed between cycles (shown in FIG. 13) and for cleaning and decontamination. The illustrated cooling block is aluminum, but other materials may be used including, but not limited to, stainless steel, composites, plastics with magnetic inserts, and other suitable materials and configurations. [0082] In operation and use, the cytometric system of the present invention can be used for counting, measuring, examining, and sorting microscopic particles in a sample. Before operation, isolation chamber 47 must be configured with the components required by the specific application. The isolation chamber includes a hinged door or window 109 sealed to an opening in the front of the chamber. The necessary components can be loaded into and affixed within the isolation chamber through the door opening in a conventional manner.

[0083] The internal components are plugged into interface panel 79 using connecting wiring and tubing. The external components plug into respective points in the interface panel. Thus, the internal components can be connected and disconnected without exposing the interior environment.

[0084] After the cytometric system is set up and the desired configuration is in place in the isolation chamber, the isolation chamber door is closed and sealed. Next, but before running a cycle, the isolation chamber is cleaned in conventional fashion with vaporous cleaner or the like. Thereafter, the isolation chamber is brought under vacuum.

[0085] Once isolation chamber 47 is cleaned and evacuated, it remains sealed from the ambient environment. Thus, no air or other fluids can move between the inside and outside of the isolation chamber. With the isolation chamber prepared, the internal components in the chamber can be connected to the rest of the system.

[0086] Further, in contrast to the prior art, the present invention isolates more components in the system. This has the advantage of further protecting the sample from contact and irritation, which improves the measuring results. Furthermore, isolation of components such as the sample source and sorting system protects users from exposure to the samples, hi the case of measuring or sorting blood samples or toxic reagents, user safety is an important concern. The isolation chamber protects users from splashing sample, toxic vapors, and other contact with the sample from the measuring site and sample source. [00871 Users operate the various subsystems of the cytometric system in an otherwise conventional fashion when running cycles. In the illustrated embodiment, isolation chamber 47 optionally includes a transport tube 110 configured to introduce components into the interior of the isolation chamber without opening or exposing the interior to the ambient environment. The transport tube has an interior volume and sealed ends, one of which feeds to the isolation chamber interior. In a conventional fashion, components can thus be deposited into the isolation chamber with minimal disturbance. Gloves extending into the interior allow a user to position and affix the components without opening the isolation chamber.

[0088] A further advantage of the present invention is that many of the components described above are removably housed within the isolation chamber. Thus, components can be released from inside the isolation chamber and removed through transport tube 110 without opening door 109 to the isolation chamber.

[0089] Using pinch valves 63 and interface panel 79, flow through the fluidic system can be stopped and started as desired. In contrast to the prior art, the pinch valves serve to prevent contact of the sample with components other than the non-reactive fluidics tubing.

[0090] With flow cut off through the tubing, the sample block 51 can then be removed from sample source 42 using lever 86. The fluidic tubing connected to the sample block can also be disconnected. Using the interface panel, the internal components including the fluidics tubing inside the isolation chamber can be disconnected. A user can then remove the fluidics system and replace it with clean, new fluidic tubing through the transport tube. Similarly, the sample source can be replaced or cleaned. The interface panel and transport tube eliminate the need for pass-throughs that prevent the isolation chamber from remaining clean.

[009i] In this manner the fluidics system and other components can be removed without disassembling the isolation chamber. In the alternative, the sample source and other components can be removed from the isolation chamber as described. The components can then be cleaned externally from the isolation chamber and then placed back inside or replaced with new components.

[0092] After running a cycle, the isolation chamber can be cleaned easily and effectively as described below. In the illustrated embodiment, the sample block collar 54 can be released by opening the cam assembly. Using the gloves 112 and the transport tube to introduce a cleaning tool, the collar can be cleaned with the isolation chamber sealed.

[0093] Furthermore, in the illustrated embodiment, the cooling block and collection tube holder 107 can be cleaned or replaced. Because the tube holder is magnetically- mounted to the cooling block, it can easily be removed. This has the further advantage of allowing a user to clean between the tube holder and cooling block and eliminate the cranny at the mounting location.

[0094] One will appreciate that the cytometric system shown and described eliminates the need for exposing the sample to an ambient environment. The system further reduces air currents within the isolation chamber.

[0095] One will further appreciate that the cytometric system aids in cleaning components in the isolation chamber and minimizes the collection of contaminants within. The system aids in the vaporous cleaning of the isolation chamber by spacing components from the floor and allowing a flow path therebetween.

[0096] Additionally, one will appreciate that the further isolation of the sample source and optional isolation of a portion of the particle sorting system increases the safety of users. The additional isolation of components has the further advantage of obviating the need for a clean room environment outside the isolation chamber. The isolation of the sample source, measuring site, and fluidics system also increases the accuracy of measured data.

[0097] Furthermore, the ease with which the interior components can be removed aids in portability of the cytometric system of the present invention. One will also appreciate that, in contrast to maintaining a sterile environment outside the isolation chamber or isolating the entire optical bench, the reduced size of the overall portable platform and clean room environment lowers the cost of the system.

[0098] One will appreciate that the cytometric system of the present invention may be used in a variety of applications. In general, the cytometric system may be used for the multiparametric analysis, sorting, counting, and general examination of microscopic particles where environmental isolation of the measuring site and sample are important. One will appreciate that different components can selected for isolation.

[0099] The foregoing descriptions of specific embodiments of the present invention have been presented for purposes of illustration and description. They are not intended to be exhaustive or to limit the invention to the precise forms disclosed, and obviously many modifications and variations are possible in light of the above teaching. The embodiments were chosen and described in order to best explain the principles of the invention and its practical application, to thereby enable others skilled in the art to best utilize the invention and various embodiments with various modifications as are suited to the particular use contemplated. It is intended that the scope of the invention be defined by the Claims appended hereto and their equivalents.

Claims

What is claimed:

1. A cytometric system for measuring particles, the system comprising: a portable platform supporting: an illumination table including a measuring site; a detection system for detecting at least one property of the particles in the measuring site; a fluidics system configured for transporting the particles from a sample source to the measuring site, the fluidics system including transfer tubing fluidly connecting the sample source to the measuring site; and an isolation chamber housing and environmentally sealing the illumination table and sample source from an ambient environment.

2. A cytometric system according to claim 1 , wherein the portable platform further supports a particle sorting system configured for sorting the particles below the measuring site based upon the detection of the at least one property of the particles.

3. A cytometric system according to claim 2, the particle sorting system including a particle sorting arm assembly, the sorting arm assembly including a sorting arm movably extending into the isolation chamber though a chamber slot, and including an isolation boot sealingly received around the sorting arm and sealingly affixed to the isolation chamber.

4. A cytometric system according to claim 2, the system further comprising a cooling block for cooling particles collected after sorting, the cooling block being housed within the isolation chamber.

5. A cytometric system according to claim 4, further comprising a collection tube holder magnetically-mounted to the cooling block within the isolation chamber.

6. A cytometric system according to claim 1, wherein the isolation chamber housing is pressure-sealed.

7. A cytometric system according to claim 6, wherein the isolation chamber is a glove box.

8. A cytometric system according to claim 7, the isolation chamber further including an interface panel in a side wall configured to interface between components in the isolation chamber and components external to the isolation chamber.

9. A cytometric system according to claim 8, wherein the interface panel is configured to receive banana plugs.

10. A cytometric system according to claim 1, further comprising a sample block for supporting the sample source, wherein the sample block is movably supported within the isolation chamber by a cam assembly to facilitate loading the sample source on the sample block.

11. A cytometric system according to claim 10, wherein the transfer tubing includes pinch valves mounted on the sample block to selectively control flow from the sample source to the measuring site.

12. A cytometric system according to claim 1 , further comprising a sample system including a sheath tank for providing sheath fluid to the measuring site, and including a waste tank for receiving waste fluid from the measuring site.

13. A cytometric system according to claim 1, wherein the illumination table includes at least one laser located outside of the isolation chamber and at least one illumination tube being configured to direct output from the at least one laser to the measuring site.

14. A cytometric system according to claim 13, wherein a portion of the at least one illumination tube is housed within the isolation chamber.

15. A cytometric system according to claim 14, wherein the isolation chamber includes at least one environmentally-sealed port in one wall configured for accepting the at least one illumination tube therethrough.

16. A cytometric system according to claim 13, the illumination table including a stage drive for horizontally aligning a sheath of particles in the measuring site with the laser output, wherein the stage drive includes a vertically extending knob for adjusting alignment.

17. A cytometric system according to claim 1, the detection system including at least one bank of detectors, a control unit, and a power source, wherein the at least one bank of detectors is housed within the isolation chamber.

18. A cytometric system according to claim 1 , wherein the sample source and illumination table are spaced from a bottom floor of the chamber to facilitate cleaning and decontamination of the isolation chamber.

19. A cytometric system according to claim 1 , wherein the particles are stem cells.

20. A cytometric system for measuring a sample of particles carried by a sheath fluid, the system comprising: an illumination table supporting a measuring site; an isolation chamber isolating an interior from the ambient environment; a detection system for detecting at least one property of the particles in the measuring site, the detection system including at least a bank of detectors, a camera having a collimating lens, a directing prism, and a power source; a fluidics system for transferring the sample to the measuring site, the fluidics system including at least a nozzle, sample source, and transfer tubing configured for transferring the sample from the sample source to the nozzle; and the illumination table, sample source, supply tubing, and nozzle being removably housed within the isolation chamber and environmentally isolated from an ambient environment.

21. A cytometric system according to claim 20, further including a particle sorting system configured for sorting the particles based upon the detection of at least one property of the particles in the sample.

22. A cytometric system according to claim 20, further wherein the at least one camera and at least one directing prism are housed within the isolation chamber.

23. A cytometric system according to claim 20, wherein the isolation chamber is configured to allow removal of the fluidics system without disassembly of the isolation chamber.

24. A cytometric system according to claim 20, wherein the particles to be measured are living cells.