METHOD AND APPARATUS FOR PERFORMING HEMATOLOGIC ANALYSIS USING AN ARRAY-IMAGING SYSTEM FOR IMAGING AND ANALYSIS OF A CENTRIFUGED ANALYSIS TUBE

Abstract

A method and device for analyzing a hematologic sample centrifuged within a capillary tube is provided. The device includes a tube holder, a sample imaging device, a processor, and a sample data display. The sample imaging device is operable to create a digital image of the sample within a region of the tube. The region is defined by substantially all of the radial width and axial length of the sample residing within the internal cavity of the tube in the region where the float resides after centrifugation. The sample imaging device is operable to produce signals representative of the image. The processor is adapted to produce information relating to bands of interest within the image based on the signals from the sample imaging device. The sample data display is adapted to display the results therefrom and/or a digital image of the sample within the region.

Claims

1. A method for analyzing a hematologic sample, comprising: disposing the hematologic sample within an internal compartment of a capillary tube, which internal compartment has a radial width and an axial length and a float disposed within the capillary tube; providing a centrifuge and a sample imaging device; using a processor to execute instructions stored within a memory device, which instructions cause the processor to: control the centrifuge to centrifugally spin the capillary tube containing sample about a central axis at speeds sufficient to create constituent layer separation within the sample disposed in the at least one capillary tube; control the sample imaging device to create a digital image of a sample region of the sample disposed within the tube, which sample region extends across all of the radial width and axial length of the internal compartment of the tube in a region of the tube where the float resides after centrifugation, and produce the signals representative of the digital image; produce information relating to the digital image based on the signals from the sample imaging device; and control a sample data display to display the information relating to the digital image of the sample region.

2. The method of claim 1, wherein the capillary tube is attached to a platen portion of the centrifuge in a position where the capillary tube extends radially outward from the central axis.

3. The method of claim 1, wherein the sample imaging device includes a digital camera.

4. The method of claim 1, wherein the instructions when executed by the processor, cause the processor to control the sample data display to display the digital image of the sample region.

5. The method of claim 4, wherein the instructions when executed by the processor, cause the processor to control the sample data display to display superimposed graphic markings based on an analysis of the sample over the digital image to illustrate band boundaries.

6. The method of claim 5, wherein the graphic markings are superimposed to identify at least one of a bottom of the tube, a bottom of the float, a red blood cell/granulocyte interphase, a granulocyte/lymphocyte and monocyte interphase, a lymphocyte and monocyte/platelet interphase, a platelet/plasma interphase, a top of the float, or a plasma/air interphase.

7. The method of claim 1, wherein the information relating to the image based on the signals from the sample imaging device includes complete blood count information.

Description

BRIEF DESCRIPTION OF THE DRAWINGS

[0012] FIG. 1 is a schematic diagram of the present invention hematology analysis device.

[0013] FIG. 2 is a schematic diagram of a capillary tube.

[0014] FIG. 3 is an enlarged partial view of a tube such as that shown in FIG. 2.

DETAILED DESCRIPTION OF THE INVENTION

[0015] Referring to FIGS. 1-3, a blood sample for analysis within the QBC hematology system is typically obtained either from a venous or capillary sample, centrifuged in a simple, small dedicated centrifuge which may be either battery powered or AC powered. U.S. Pat. Nos. 4,027,660; 4,683,579 and 6,441,890, each of which is hereby incorporated by reference in its entirety, describe methods and apparatus for hematological analysis using a capillary tube and a space occupying insert that floats on the centrifuged red blood cells thereby expanding the surrounding buffy coat and permitting the measurement and quantization of the blood's layers. The capillary tube 10 includes a body that extends between a closed bottom 12 and an open top 14. In some embodiments, the closed bottom may be vented to allow the escape of gas. The open top 14 provides access to an internal cavity 16 that has a radial width 18 and an axially extending length 20. In those embodiments where the tube 10 is cylindrical, the radial width 18 is the inner diameter of the tube 10. The present invention is not limited to use with any particular type of capillary tube. U.S. Pat. No. 4,027,660, for example, describes a QBC style capillary tube operable to contain a fluid sample and a volume occupying mass 22 (hereinafter referred to as a float), and the information available by virtue of the relative positioning of the float 22 within the sample after centrifugation. U.S. Pat. No. 6,444,436 describes a different style of capillary tube that can be used with the present invention; e.g., one having a polynomial (e.g., rectilinear) cross-sectional geometry. FIGS. 2 and 3 of the present application diagrammatically illustrate a capillary tube 10 with a sample and a float 22 disposed in the internal cavity 16 of the tube 10. The centrifuged sample disposed in the tube 10 illustrates the constituent bands 24 (24a, 24b, 24c, 24d, 24e) and the band boundaries 25 (25a, 25b, 25c, 25d) therebetween. U.S. Pat. Nos. 4,683,579 and 6,441,890 describe automated devices for reading the centrifuged sample by way of an axially extending linear scan of a limited portion of the sample disposed within the QBC tube, which limited linear portion is disposed at a particular circumferential position of the tube 10.

[0016] The present invention analysis device operates with a capillary tube 10 such as those provided within a QBC hematology system; i.e., a tube 10 filled with a sample that has been centrifuged to produce the separated constituent layers (i.e., bands) 24 within the sample. One embodiment of the present analysis device 28 includes a housing 30 containing a tube holder 32, a sample imaging device 34, a processor 36 adapted to produce information relating to bands 24 of interest within the image based on the signals from the sample imaging device 34, a sample data display 38, and may include an operator input device 40 that enables the operator to enter relevant patient information.

[0017] In some embodiments, the analysis device 28 further includes a centrifuge 42 with a platen 44 configured to hold one or more capillary tubes 10 in a position where the tubes 10 extend radially outward from a central axis. In these embodiments, the analysis device 28 can perform both the centrifugation and the image analysis. The centrifuge 42 is operable to centrifugally spin the tube 10 containing the sample about the central axis at speeds sufficient to create constituent layer separation within the sample disposed in the tube 10. In these embodiments, the platen 44 is an example of a tube holder 32. In other embodiments, the tube holder 32 may be independent of the centrifuge 42.

[0018] The sample imaging device 34 includes a digital camera operable to image substantially all of the radial width 18 and axial length 20 of the sample residing within the internal cavity 16 of the tube 10 in the region 46 (see FIG. 3) where the float 22 resides after centrifugation in a single image, and to produce signals representative of the image. In the preferred embodiments, the sample imaging device 34 is operable to image a region 48 comprising substantially all of the radial width 18 and axial length 20 of the sample within the tube 10 in a single image, and to produce signals representative of the image. Alternately, two or more cameras can be used to image separate portions of the tube 10, which portions are contiguous with one another. The images of the contiguous regions can be subsequently combined and analyzed or are separately analyzed. Either the digital camera itself, or an independent light source within the sample imaging device 34, provides sufficient lighting so that bands 24 of interest within the centrifuged sample may be differentiated within the sample image. The optical resolution of the camera must be sufficient to provide adequate clarity within the image for the analysis at hand; e.g., to differentiate bands 24 of interest. As indicated above, the sample imaging device 34 may be incorporated into a QBC tube type reader, or may be an independent device (e.g., a portable digital camera, a cell phone camera, etc.) configured for use with such a reader. An example of an acceptable digital camera is a Bayer-type matrix color camera. If, for example, a standard Aptina five megapixel color camera chip with a frame width of 2592 pixels is used, it can produce a theoretical image resolution of 0.02 mm, which is acceptable for most analyses. If a color camera is used, color filters and different illumination types are likely not required. A grey scaled camera may also be used because the separated buffy coat layers have different light scattering properties and may therefore be detected using a black and white camera, although this measurement is less robust and requires more controlled illumination. The sample imaging device 34 may be described as an area-array imaging device because it images substantially all of the radial width 18 and axial length 20 of the sample within the interior cavity 16. If a plurality of cameras is used within the present sample imaging device 34, the images they produce are contiguous with one another thereby permitting the plurality of images to be combined into a single representative image. The linear scan devices of the prior art, in contrast, are limited to producing narrow linear segments that do not extend across the full radial width 18, and the circumferential linear segments are not contiguous with each other. As a result, the circumferentially positioned linear segments cannot be combined into a single representative image. Examples of acceptable independent light sources include white and/or blue LEDs, operable either in a steady state mode or in the case of the QBC STAR type reader, in a pulsed mode. The relative blue spectrum of a white LED or the inclusion of a separate blue LED can excite the fluorescence of a dye such as Acridine Orange in the tube 10.

[0019] The processor 36 is adapted (e.g., programmed) to perform several tasks, including: a) controlling the sample imaging device 34 based on the analysis at hand; b) controlling the centrifuge 42 for those embodiments that include one; c) receiving and acting on operator input entered through the operator input device 40; and d) producing information relating to bands 24 of interest within the image based on the signals from the sample imaging device 34. The extent of the information relating to the bands 24 can vary depending upon the embodiment of the device 28. For example, the processor 36 may be adapted to provide information relating to the adequacy of the sample image, and/or with algorithmic capability that is operable to analyze the signals representative of the sample image and produce data (e.g., CBC, hematocrit, WBC count, etc.) relating thereto based on characteristics of the different bands 24 within the centrifuged sample. In some applications, the processor 36 can be adapted to produce graphic markings based on the analysis of the sample that can be superimposed over the sample image when displayed to illustrate the calculated band boundaries 25 relative to the sample image. Using the analysis of a blood sample as an example, graphic markings can be used to identify features such as the: a) bottom of the tube 10; b) bottom of the float 22; c) red blood cell/granulocyte interphase; d) granulocyte/lymphocyte and monocyte interphase; e) lymphocyte and monocyte/platelet interphase; f) platelet/plasma interphase; g) top of the float 22; h) plasma/air interphase; etc.

[0020] The sample data display 38 is in communication with the processor 36 and includes a display screen. The display screen is an electronic screen (e.g., flat screen LED, LCD, etc.) operable to display the calculated results and/or a digital image of the sample residing within the centrifuged sample with sufficient optical resolution so that the image can be evaluated by a technician operator to provide the information pertaining to the bands 24 of interest within the centrifuged sample. In those embodiments that include an operator input device 40 (e.g., key pad, touch screen, etc.), the operator input device 40 allows the operator to enter relevant patient or other information, if desired. The sample display 38 may be integral with the housing 30, or it may be an independent device in communication with the processor 36. For example, universal monitors are often used in medical facilities, which monitors have the capability of displaying data from more than one analysis device. In such an application, the data to be displayed may be viewed on an integral display screen and/or a remotely located display device in communication with the processor 36.

[0021] In some embodiments, the analysis device 28 includes a communication port 50 for sending the signals representative of the sample image to a remote location. The communication port 50 can be a hardwire port for communicating by hardwire connection to a remote site, or it can be a wireless communication connection (e.g., similar to that used in a wireless phone).

[0022] In some embodiments, fiduciary marks 52 (i.e., calibration, measurement marks, etc.) may be placed on or in the capillary tube 10, or the tube holder 32, or on a measuring device positioned adjacent the tube 10 (e.g., a ruler) to facilitate geometric and/or optical calibration and thereby account for any image distortion introduced by the camera. In those instances where the fiduciary marks are placed on or in the tube, a particularly useful embodiment is one wherein the marks are positioned relative to the internal cavity to permit geometric evaluation of sample within the internal cavity. In those instances where fiduciary marks 52 are disposed on a measuring device positioned adjacent the tube 10, the measurement device can measure along an axis that is maintained parallel to the lengthwise axis (e.g., axial direction) of the tube 10. In such embodiments, the measurement device is preferably in close proximity (e.g., in the same focal plane) as the sample tube 10. Alternately, a look-up-table can be provided by factory calibration to serve this function. During the image processing and analysis steps, the calibration information can be used to ensure correct length measurements of the tube 10 features, regardless of their position in the image frame or distance from the camera and can compensate for instrument-to-instrument differences.

Operation:

[0023] A fluid sample (e.g., whole blood) is collected from a patient and deposited into a capillary tube 10 such as those used in the QBC hematology system for subsequent centrifugation. As indicated above, the centrifuge may be independent of, or incorporated into, the analysis device 28. The sample is centrifuged for a period of time adequate to create constituent layer separation within the sample disposed in the tube 10, and the representative bands 24 associated therewith. The centrifuged sample is then imaged using the sample imaging device 34. The image includes substantially all of the radial width 18 and axial length 20 of the sample residing within the internal cavity 16 of the tube 10 in the region where the float 22 resides after centrifugation. Because capillary tubes 10 are not always filled with the exact same volume of fluid sample, the sample imaging device 34 preferably images the region 48 of the tube 10 from the top meniscus to the bottom of the red blood cell layer. It is desirable, but not required, that the bottom of the tube 10 be imaged as well. If the sample being imaged is disposed within a STAR type QBC tube, for example, the total length between the tube bottom to the tube top fill position is approximately 53 mm. The distance from the tube top fill position to the bottom of the float 22 in most instances is about 37 mm. In those device 28 embodiments that include a centrifuge, the sample may be centrifuged and the centrifuge subsequently stopped or slowed to a very low RPM prior to the imaging. The sample imaging device 34 produces signals representative of each image and communicates those signals to the processor 36.

[0024] The images signals are subsequently analyzed within the processor 36 using image processing algorithms to isolate and analyze the bands 24 of interest within the sample, and in some instances relevant sections of the bands 24. Before or after the image signals are analyzed, the image signals may be sent to the sample data display 38 for evaluation by the operator. The ability to have an operator visually evaluate an image that includes substantially all of the radial width 18 of the sample within the tube 10, and a relevant portion of the axial length 20 of the sample is a substantial advancement of the technology. A person of skill in the art will recognize that no automated system can account for all potential variables within the sample image. For example, during the centrifugation process, there is a chance that sample will exit the capillary tube 10 and pass into the retaining tube of the centrifuge. In such instances, the released sample can contaminate the exterior of the capillary tube 10 and inhibit accurate analysis. Similarly, a misplaced tube label or debris deposited on the exterior of the capillary tube 10 during handling can also inhibit or prevent accurate analysis. In these instances, the ability of the present device 28 to produce a single substantially complete image of the centrifuged sample will enable the operator to identify such potential problems and take appropriate action. As another example, the image available with the present device 28 will also enable the operator to evaluate other aspects of the sample image for potential problems; e.g., overall image quality, accuracy of sample coloration, the degree to which a blood sample may be lipemic or icteric, etc. In those applications where the operator evaluates the image after processing and boundary markings are assigned by the processor 36, the operator can evaluate whether the assigned boundary markings are accurately positioned relative to the sample image. Hence, the ability to have an operator visually evaluate an image using the present device 28 provides considerable quality controls to the analysis process. It should be emphasized that the present instrument, as described herein, may be used in locations where trained operators are present, and also locations where no trained operators are present (e.g., a CLIA-waived environment). In the latter type location, the sample images captured by the present device 28 may be sent to a remotely located trained operator for analysis. If it is not possible to have a trained operator review the image and/or results within a predetermined period of time, the present device 28 may be programmed to prevent the release of any data if the sample image has any detectable anomalies. A visual image analysis is preferable in that the criteria for analysis rejection can be loosened, but a purely automated analysis (e.g., that checks for anomalies) is preferable to no analysis at all.

[0025] The extent to which the present device 28 images the centrifuged sample within the tube 10 makes possible another quality control mechanism. As indicated above, the present device 28 images substantially all of the radial width 18 and a significant portion of the axial length of the centrifuged sample. In some instances, the radial portion of the image can be expanded to a point outside of the capillary tube 10 to include other imageable features such as calibration markers or areas. The image characteristics associated with the regions outside of the capillary tube 10 can be compared against the characteristics of the region inside the tube 10. Inconsistencies identified by the comparison of the characteristics (e.g., brightness) can be used to evaluate the accuracy of the image. This type of quality control is not possible using the prior art reading devices that utilize a linear scanning device, which has essentially only a one pixel width.

[0026] Although the invention has been described and illustrated with respect to exemplary embodiments thereof, the foregoing and various other additions and omissions may be made therein and thereto without departing from the spirit and scope of the present invention.