This invention relates generally to the flow cytometer field, and more specifically to a flow cytometer system with a flow cell body, a flow channel, and a sample injection probe.
A typical flow cell for a flow cytometer system, which includes a flow channel, is composed of multiple pieces of fused silica that must be individually cast or cut and later assembled. The typical flow channel is susceptible to clogs and bubbles. A clog, which prevents flow of the sample fluid, may be caused by sample debris, conjugated or clustered cells, or other substances inserted into the flow path of the flow cytometer. Bubbles may interfere with the optical interrogation of the sample as it passes through the interrogation zone. Both clogs and bubbles within the flow channel can render experimental data useless, which in turn leads to repetitive experiments, increased costs, and lost time associated with the maintenance and operation of the flow cytometer.
Thus, there is a need for a flow cell that provides for improved construction and integration of its component parts, as well as a flow cytometer system that reduces the likelihood of clogs and bubbles in the flow channel. This invention provides such an improved and useful flow cytometer.
The following description of the preferred embodiments of the invention is not intended to limit the invention to these preferred embodiments, but rather to enable any person skilled in the art of flow cytometry to make and use this invention.
As shown in
1. Flow Cell Body, Flow Channel, and Sample Injection Probe
The flow cell body 110 of the preferred embodiment functions to contain, protect, and align the components of the flow cytometer system. As shown in
The flow cell body 110 is preferably manufactured according to methods known in the art of manufacture, including for example CNC machining and injection molding or any combination thereof. The method of manufacture of the flow cell body 110 of the preferred invention includes the steps of providing a material, and disposing a receiving channel 116 in the material such that the receiving channel 116 is appropriately sized for receiving and holding the flow channel 112. Suitable materials include polycarbonate, although any suitable metal, plastic, alloy, or composite material can be readily substituted for material. The receiving channel 116 is preferably manufactured such that it provides an opening through which the flow channel 112 is radially exposed for the interrogation of the samples within the flow channel 112.
The flow channel 112 of the preferred embodiment is coupled to the flow cell body 110 and functions to conduct and focus sample fluid through an interrogation zone, where the sample material is analyzed. As shown in
The sample injection probe (SIP) 114 of the preferred embodiment is removably coupled to the flow cell body 110 and functions to provide a uniform flow of sample fluid to the flow channel 112. As shown in
In a variation of the preferred embodiment, the flow cytometer system includes a SIP body 118, as shown in
The SIP body 118 may further define a circumferential groove 150 that functions to hold a back-up o-ring 158, or any other suitable element that will increase friction between the SIP body 118 and the flow cell body 110 at a point, that functions to create a seal and/or to create a removable press fit connection between the flow cell body 110 and the SIP body 118. The flow cytometer system may further include a plurality of o-rings. As shown in
2. Assembly of the Flow Cytometer System
To ensure accurate analysis of the sample material, the flow channel 112 is preferably correctly aligned with the flow cell body 110. Due to the small size of the flow channel 112 (it is typically a capillary tube less than 0.3 mm in diameter), it is difficult to accurately radially align the capillary tube with the flow cell body 110 during assembly of the flow cell for a flow cytometer system. The flow cytometer is preferably assembled by a method with a device that facilitates the radial alignment of a capillary tube in a flow cell. The device is preferably the assembly fixture 10 of the preferred embodiments, but may alternatively be any suitable device used in any suitable method.
As shown in
The assembly fixture 10 of the preferred embodiments is an article of manufacture, preferably made out of a metal such as brass. The assembly fixture 10 may alternatively be made out of any suitable rigid material such as stainless steel, plastic or composite. The assembly fixture is preferably constructed of a single unitary piece, which may be fabricated through any known methods such as CNC machining, injection molding, and the like.
The capillary receptacle 12 of the preferred embodiments functions to receive a capillary tube for assembly. As shown in
As shown in
The alignment element 14 preferably includes at least one of a nozzle region mating interface 20 and SIP body mating interface 22. The nozzle region mating interface 20 is preferably located at the top portion of the assembly fixture 10 and is substantially cylindrically shaped. The nozzle region mating interface 20 is preferably dimensioned such that it fits tightly with the flow cell nozzle region of the flow body. The SIP body mating interface 22 is preferably a tapered or conical geometry that is configured for precision alignment with the portion of the flow cell body 110 with which the SIP body mating interface 120 aligns. The dimensions of the SIP body mating interface 22 are preferably the same or similar to the mating interface 120 of the SIP body 118. The use of a tapered or conical geometry assures both a particular radial and axial mating between the flow cell and the assembly fixture. Other suitable dimensions may, however, be used.
The coupling element 16, as shown in
The assemble fixture 10 of the preferred embodiment may further include a handle 18, as shown in
The assembly fixture 10 of the preferred embodiments is preferably used to assemble a flow cell. The method of assembling the flow cytometer system of the preferred embodiments includes providing a flow cell body 110, a capillary tube (flow channel 112), and an assembly fixture 10; coupling the capillary tube to the assembly fixture 10 in the capillary receptacle 12 of the assembly fixture 10; coupling the assembly fixture 10 to the flow cell body 110 and attaching the capillary tube to the flow cell body 110; and removing the assembly fixture 10 from the flow cell body 110. The method is preferably designed for the assembly of the flow channel 112 and the flow cell body 110 of the preferred embodiments. The method, however, may be alternatively used in any suitable environment and for any suitable reason.
The step that recites providing a flow cell body 110, a capillary tube (flow channel 112), and an assembly fixture 10, functions to provide the elements of the flow cytometer system that will be coupled together. The step that recites coupling the capillary tube to the assembly fixture 10 in the capillary receptacle 12 of the assembly fixture 10, functions to place the capillary tube in the capillary receptacle such that the capillary tube is radially aligned by the walls of the receptacle and a portion of the capillary tube remains exposed beyond the assembly fixture 10 and is axially aligned.
The step that recites coupling the assembly fixture 10 to the flow cell body 110 and attaching the capillary tube to the flow cell body 110, functions to couple the assembly fixture 10 with the correctly aligned capillary tube into the flow cell body 110, as shown in
The step that recites removing the assembly fixture 10 from the flow cell body 110, functions to uncouple the coupling element 16 and remove the assembly fixture 10, leaving a properly assembled capillary tube in place within the flow cell body 110.
Although omitted for conciseness, the preferred embodiments include every combination and permutation of the assembly fixture 10, the capillary receptacle 12, the alignment element 14, the coupling element 16, the handle 18, and any method of assembling the capillary tube within a flow cell body 110 using the assembly fixture 10.
3. Other Aspects of the Invention
In a variation of the preferred embodiment, the flow cytometer system includes a bubble purge port 126, as shown in
In another variation of the preferred embodiment, the flow cytometer system includes a hydrodynamic focusing region 128, as shown in
In another variation of the preferred embodiment, the flow cytometer includes a SIP cleaning port 132, as shown in
In another variation of the preferred embodiment, the flow cytometer includes a pressure monitoring tube 134, as shown in
In another variation of the preferred embodiment, the flow cytometer includes a SIP filter 136, as shown in
In another variation of the preferred embodiment, the flow cytometer includes a sample vial mechanism 138, as shown in
In another variation of the preferred embodiment, as shown in
In another variation of the preferred embodiment, as shown in
In another variation of the preferred embodiment, as shown in
As a person skilled in the art of flow cytometry will recognize from the previous detailed description and from the figures and claim, modifications and changes can be made to the preferred embodiments of the invention without departing from the scope of this invention defined in the following claim.
This application claims the benefit of U.S. Provisional Application No. 60/864,646 filed 7 Nov. 2006 and entitled “FLOW CELL FOR A FLOW CYTOMETER SYSTEM”, which is incorporated in its entirety by this reference.
Filing Document | Filing Date | Country | Kind | 371c Date |
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PCT/US2007/083991 | 11/7/2007 | WO | 00 | 1/19/2010 |
Publishing Document | Publishing Date | Country | Kind |
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WO2008/058217 | 5/15/2008 | WO | A |
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