Patent Description:
The present disclosure is directed to methods and apparatus for characterizing a biological specimen, and, more particularly to methods and apparatus for determining the presence of an interferent in the biological specimen and/or segmenting the biological specimen.

Automated testing systems may conduct clinical chemistry or assays using one or more reagents to identify an analyte or other constituent in a biological specimen such as blood serum, blood plasma, and the like. For convenience and safety reasons, these specimens may be contained in specimen containers (e.g., blood collection tubes).

Improvements in automated testing technology have been accompanied by corresponding advances in pre-analytical sample preparation and handling operations such as batch preparation, centrifugation of the biological specimen to separate specimen constituents, cap removal to facilitate specimen access, automated aliquoting, and the like by automated pre-analytical specimen preparation systems, which may be part of a Laboratory Automation System (LAS). The LAS may automatically transport specimens contained in the specimen containers, and received on carriers, to a number of pre-analytical specimen processing stations recited above, as well as to analytical stations containing clinical chemistry analyzers and/or assay instruments (collectively referred to as "analyzers" herein).

LASs may handle all different sizes and types of specimen containers, which may contain barcode labels. The barcode label may contain an accession number that may be correlated to demographic information that may be entered into a hospital's Laboratory Information System (LIS) along with test orders and other information. An operator or robot may place the barcode-labeled specimen containers onto the LAS system, and the LAS system may automatically transport the specimen containers for such pre-analytical operations, all prior to the specimen actually being subjected to clinical analysis or assaying by the one or more analyzers.

For certain tests, an amount of a serum or plasma portion of the specimen obtained from whole blood by fractionation (e.g., centrifugation) may be aspirated and used. In some instances, a gel separator may be added to the specimen container to aid in physically separating a settled blood portion from the serum or plasma portion. In some embodiments, after fractionation and a subsequent de-capping process, the specimen container may be transported to an appropriate analyzer on the LAS that may extract via aspiration with a pipette, serum or plasma portion from the specimen container and combine the serum or plasma portion with one or more reagents, diluents, and possibly other substances in a reaction vessel (e.g., cuvette). Analytical measurements may then be performed, often using a beam of interrogating radiation, for example, or by using photometric or fluorometric absorption readings, or the like. The measurements allow for the determination of end-point or rate or other values, from which the concentration of analyte or other constituent may be determined using well-known techniques.

In some specimen testing apparatus, the specimen containers may be of different size and also the total amount of specimen contained therein as well as the relative amounts of settled blood portion and serum or plasma portion may vary substantially from specimen container to specimen container. This may lead to uncertainty in terms of robot and pipette movement and positioning.

Furthermore, the presence of one or more interferent in the specimen as a result of sample processing or patient disease condition may possibly adversely affect the accuracy of the test results of the analyte or constituent measurement obtained from the one or more analyzers. For example, the presence of hemolysis (hereinafter "H"), icterus (hereinafter "I"), and/or lipemia (hereinafter "L") which are collectively H, I, and/or L (hereinafter referred to as "HIL") may affect specimen testing results. However, determining HIL may be very computationally intensive. <CIT> discloses an apparatus for testing for the presence of an interferent in a sample, such as hemolysis, icterus, and/or lipemia in a serum portion of a blood sample. The apparatus comprises one or more digital RGB cameras for capturing pixelated images of a centrifuged blood sample in a container, and a computer for analyzing the pixelated images to classify pixels in the pixelated image as being liquid or non-liquid, defining a liquid region based upon the classification of the pixels, and determining a presence of hemolysis, icterus, and/or lipemia within the liquid region. <CIT> describes a hyperspectral imaging system for determining protein and hemoglobin in urine.

Because of problems encountered when different-sized specimen containers are used in the LAS, the need to know the location of and/or amounts of serum or plasma present, as well as the need to pre-screen for HIL in the specimen to be analyzed, there is an unmet need for a computationally-efficient method and apparatus adapted to readily and automatically image and analyze such specimens.

According to a first aspect of the invention, a characterization apparatus as defined in claim <NUM> is provided. The characterization apparatus comprises: an imaging location configured to receive a specimen container containing a specimen, wherein the specimen includes a settled blood portion and a serum or plasma portion; a light source configured to provide lighting of the imaging location; a hyperspectral image capture device comprising a spectrally-resolving element and a spectral image capture device, the hyperspectral image capture device configured to generate and capture a spectrally-resolved image of a portion of the specimen container and specimen at the spectral image capture device; and a computer configured and operable to process the spectrally-resolved image received at the spectral image capture device and determine a presence or absence of an interferent, the interferent being hemolysis, icterus and/or lipemia and at least one of segmentation of the specimen or segmentation of the specimen container and the specimen.

In another aspect of the invention, a specimen testing apparatus as defined in claim <NUM> is provided. The specimen testing apparatus comprises: a track; a carrier moveable on the track and configured to support a specimen container containing a specimen, wherein the specimen includes a settled blood portion and a serum or plasma portion; and the characterization apparatus of the first aspect, located on the track, wherein the imaging location of the characterization apparatus is configured to receive the specimen container containing the specimen carried by the carrier.

According to another aspect of the invention, a method of characterizing a specimen container and/or a specimen as defined in claim <NUM> is provided.

The method comprises: providing the specimen container containing the specimen at an imaging location, wherein the specimen includes a settled blood portion and a serum or plasma portion; providing a hyperspectral image capture device configured to capture an image at the imaging location; providing one or more light sources configured to provide illumination of the imaging location; illuminating the specimen container containing the specimen provided at the imaging location with the one or more light sources; capturing a spectrally-resolved image of a portion of the specimen container and specimen with the hyperspectral image capture device; and processing the spectrally-resolved image to determine a presence or absence of an interferent, the interferent being hemolysis, icterus and/or lipemia and at least one of: segmentation of the specimen, or segmentation of the specimen container and the specimen.

Still other aspects, features, and advantages of the present disclosure may be readily apparent from the following description by illustrating a number of example embodiments, including the best mode contemplated for carrying out the present invention. The drawings and descriptions are to be regarded as illustrative in nature, and not as restrictive.

The drawings, described below, are for illustrative purposes only and are not necessarily drawn to scale. The drawings are not intended to limit the scope of the disclosure in any way.

Embodiments of the present disclosure provide methods and apparatus adapted to image and to characterize a specimen contained in a specimen container using hyperspectral imaging. In one or more embodiments, the end result of the characterization method may be the quantification of the specimen contained in the specimen container. For example, the quantification may include characterizing of a location of an upper extent, a lower extent, and even a depth of the serum or plasma portion, and/or the location of an upper extent, lower extent, or even a depth of the settled blood portion of a fractionated blood specimen. These segmented values may be used in later processing. For example, these segmenting values may be used to determine if sufficient amount of the serum or plasma portion is present for the ordered testing, for determining disease state of the patient (e.g., a ratio between the serum or plasma portion and the settled blood portion), and/or for more exact probe tip placement to avoid aspirating air or settled blood portion.

Furthermore, according to one or more embodiments, the present apparats and methods including hyperspectral imaging may be used to determine one or more geometric characteristics of the specimen container. For example, dimensional characteristics of the specimen container may be determined, such as container height, and/or height of a cap. These dimensional characteristics may be used to properly guide the positioning of the probe (otherwise referred to as a "pipette") during a subsequent aspiration process. Such information may be used to avoid contact or crashes of a robot gripper or probe with the specimen container during maneuvers with the robot gripper and/or probe.

In some embodiments, the characterization apparatus and methods including hyperspectral imaging may capture specimen image intensities that may be used for making a determination of a presence of an interferent, such as the presence of hemolysis (H), icterus (I), and/or lipemia (L) in the serum or plasma portion of the specimen or for making a determination that the specimen is normal (N). The present method and apparatus including hyperspectral imaging is computationally effective for each of segmentation and/or interferent detection.

The specimen, as described herein, is collected in a specimen container, such as a blood collection tube, and may include a settled blood portion and a serum and plasma portion after separation (e.g., fractionation using centrifugation). The settled blood portion is made up blood cells such as white blood cells (leukocytes), red blood cells (erythrocytes), and platelets (thrombocytes), which are aggregated and separated from the serum or plasma portion. The settled blood portion is found settled/packed at the bottom part of the specimen container. The serum or plasma portion is the liquid component of blood that is not part of the settled blood portion. It is found above the settled blood portion. Plasma and serum differ primarily in the content of coagulating components, primarily fibrinogen. Plasma is the un-clotted liquid, whereas serum refers to blood plasma that has been allowed to clot, either under the influence of endogenous enzymes or exogenous components. In some specimen containers, a gel separator (e.g. a small plug) may be used, which positions itself between the settled blood portion and the serum or plasma portion during fractionation. It serves as a barrier between the two portions and minimizes remixing thereof.

In accordance with one or more embodiments, the characterization methods described herein may be carried out as pre-analytical testing or pre-screening methods. For example, in one or more embodiments, the characterization methods may be carried out prior to the specimen being subjected to analysis (clinical chemistry or assaying) on one or more analyzers. In one or more embodiments, the characterization of the specimen may be determined at one or more characterization apparatus including one or more hyperspectral image capture device. Each of the one or more hyperspectral image capture device is configured to capture one or more spectral images of the specimen container and specimen from a lateral viewpoint. During image capture, the specimen container and specimen may be illuminated. The illumination may be by one or more light panel assemblies. In particular, the illumination may be by provided by back lighting with one or more light panel assembly in some embodiments. In other embodiments, the illumination may be by provided by front lighting with one or more light panel assemblies. The hyperspectral image capture device captures spectral images that are processed to characterize the specimen container and/or specimen, including determining whether an interferent, such as HIL is present in the specimen.

In other embodiments, the methods and apparatus including hyperspectral imaging may be used to identify other characteristics of the specimen container, such as the container type (via identification of height thereof), and may further characterize the cap type, and/or the cap color.

If after characterization by the methods and apparatus described herein, the serum or plasma portion is found to contain one or more of H, I, or L, the specimen may be subjected to further processing. For example, the specimen container containing specimen may be taken to another station (e.g., a remote station) for further processing, or for additional characterization of indexes for, H, I, or L. After such further processing, the specimen may be allowed, in some embodiments, to continue on and undergo routine analysis by the one or more analyzers. In other cases, the specimen may be discarded and redrawn. If the pre-screening finds that the specimen is normal (N), then the specimen may be directly routed to undergo the ordered analysis by the one or more analyzers.

In one or more embodiments, a characterization apparatus is configured to carry out the image capture as part of the LAS where a track transports the specimen to one or more analyzers, and the characterization apparatus may be provided at any suitable location on, or along, the track. For example, characterization apparatus may be located at a loading station, on the track, or elsewhere alongside of the track, so that the specimen and specimen container can be characterized before being received at the one or more analyzers. However, to be clear, the characterization apparatus including hyperspectral imaging may receive the specimen container including specimen other than on a track, and the specimen container including specimen may be loaded and unloaded therefrom either manually or by a robot gripper.

The characterization method may be accomplished in one or more embodiments by using hyperspectral imaging wherein a spatial image of a portion of the specimen and specimen container is optically transformed into a spectral image and received by a spectral image capture device. The transformation from the spatial regime to the spectral regime may be accomplished by an arrangement of lenses, a spectrally-resolving element, such as a prism or grating, and a slit aperture. The slit aperture provides that only a sub-portion of the specimen container is imaged, such as a small central portion of the specimen container width. In some embodiments, the lighting source may be a broad band source. For example, the broad band source may emit a wavelength range from <NUM> to <NUM>. However, some embodiments, the light source may be a white light source having a wavelength emission range of <NUM> to <NUM>.

The spectral signature received by the spectral image capture device for every vertical incremental portion is processed by a computer. In one embodiment, the processing may provide segmentation information about the specimen and/or specimen container. For example, the segmentation of the specimen may determine a vertical location of one or more of: a serum or plasma portion in the specimen, a settled blood portion of the specimen, a gel separator, air in the specimen container, and a cap. Additionally or optionally, the segmentation of the specimen may determine a vertical location of one or more of: a tube-cap interface, a liquid-air interface, a serum-blood interface, a serum- gel interface, and a blood-gel interface. Moreover, once the vertical serum or plasma portion location is known in the image, a determination of HIL or N can be made for that segment, while ignoring the other segmented regions.

Further details of the characterization methods, characterization apparatus, and specimen testing apparatus including one or more characterization apparatus are further described with reference to <FIG> herein.

<FIG> shows a specimen testing apparatus <NUM> capable of automatically processing multiple ones of the specimen containers <NUM> (e.g., blood collection tubes - see <FIG>). The specimen containers <NUM> may be contained in one or more racks <NUM> at a loading area <NUM> prior to transportation to, and analysis by, one or more analyzers (e.g., first, second, and third analyzer <NUM>, <NUM>, <NUM>, respectively) arranged about the specimen testing apparatus <NUM>. It should be apparent that more or less numbers of analyzers can be used. The analyzers may be any combination of clinical chemistry analyzers and/or assaying instruments, or the like. The specimen containers <NUM> may be any transparent or translucent container, such as a blood collection tube, test tube, or other clear glass or plastic container configured to contain a specimen <NUM>.

Typically, a specimen <NUM> (<FIG>) to be imaged may be provided in the specimen containers <NUM>, which may be capped with a cap <NUM>. The caps <NUM> may have different heights and/or colors (e.g., red, royal blue, light blue, green, grey, tan, or yellow, or combinations of colors), which may have meaning in terms of what test the specimen container <NUM> is used for, the type of additive that is contained therein, and the like. Other colors may be used. It may be desirable to image the cap <NUM> to characterize information about the cap <NUM> so that it can be used to cross check with test orders to ensure the right specimen container <NUM> was used for the test that was ordered.

Each of the specimen containers <NUM> may be provided with identification information 218i (i.e., indicia), such as a barcode, alphabetic, numeric, alphanumeric, or combination thereof that may be machine readable at various locations about the specimen testing apparatus <NUM>. The identification information 218i may indicate, or may otherwise be correlated, via a Laboratory Information System (LIS) <NUM>, to a patient's identification as well as tests to be accomplished upon the specimen <NUM>, or other information, for example. Such identification information 218i may be provided on a label <NUM> adhered to, or otherwise provided on a lateral side of the specimen container <NUM>. The label <NUM> may not extend all the way around the specimen container <NUM>, or all along a height of the specimen container <NUM>. In some embodiments, multiple labels <NUM> may be adhered and may slightly overlap each other. Accordingly, although the label <NUM> may occlude a view of some portion of the specimen <NUM>, some portion of the specimen <NUM> may still be viewable from at least one lateral viewpoint.

As best shown in <FIG>, the specimen <NUM> may include a serum or plasma portion 212SP and a settled blood portion 212SB contained within the tube <NUM>. Air <NUM> may be provided above the serum and plasma portion 212SP and a line of demarcation between the air <NUM> and the serum and plasma portion 212SP is defined herein as a liquid-air interface LA. A line of demarcation between the serum or plasma portion 212SP and the settled blood portion 212SB is defined herein as a serum-blood interface SB. An interface between the air <NUM> and the cap <NUM> is referred to herein as a tube-cap interface TC. A height of the serum or plasma portion 212SP is HSP and is defined as a height from the top of the serum or plasma portion 212SP to a top of the settled blood portion 212SB, i.e., from LA to SB. A height of the settled blood portion 212SB is HSB and is defined as a height from a bottom of the settled blood portion 212SB to the top of the settled blood portion 212SB at SB. HTOT in <FIG> is a total height of the specimen <NUM> and HTOT = HSP + HSB.

In cases where a gel separator <NUM> is used (see <FIG>), a height of the serum or plasma portion 212SP is HSP and is defined as the height from the top of the serum or plasma portion 212SP at LA to a top of the gel separator <NUM> at SG. The height of the settled blood portion 212SB is HSB and is defined as the height from the bottom of the settled blood portion 212SB to a bottom of the gel separator <NUM> at BG. HTOT in <FIG> is the total height of the specimen <NUM> and is defined as HTOT = HSP + HSB + height of the gel separator <NUM>. In both cases, a height of the tube HT is defined herein as the height from the bottom-most part of the tube <NUM> to a bottom of the cap <NUM>.

In more detail, specimen testing apparatus <NUM> may include a base <NUM> (e.g., a frame or structure) upon which a track <NUM> may be mounted or rest. The track <NUM> may be a railed track (e.g., mono- or multiple-rail track), a collection of conveyor belts, conveyor chains or links, moveable platforms, or any other suitable type of conveyance mechanism. Track <NUM> may be circular, serpentine, or any other suitable shape and may be a closed track (e.g., endless track) in some embodiments. Track <NUM> may, in operation, transport individual ones of the specimen containers <NUM> to destination locations spaced about the track <NUM> in carriers <NUM> (a few carriers <NUM> are labeled).

Carriers <NUM> may be passive, non-motored pucks that may be configured to carry a single specimen container <NUM> on the track <NUM>, where the track <NUM> is moveable. Optionally, carrier <NUM> may be automated including an onboard drive motor, such as a linear motor that is programmed to move about the track <NUM> and stop at pre-programmed locations, where the track <NUM> is stationary. In either case, the carriers <NUM> may each include a holder <NUM> (<FIG>) configured to hold and support the specimen container <NUM> in a defined, upright orientation. The holder <NUM> may include a plurality of fingers, leaf springs, or combinations thereof that secure the specimen container <NUM> in the carrier <NUM>, but where at least some are laterally moveable or flexible to accommodate for different sizes of specimen containers <NUM> to be received therein.

In some embodiments, carriers <NUM> may exit from the loading area <NUM> having one or more racks <NUM> staged thereat. In some embodiments, loading area <NUM> may serve a dual function of allowing offloading of the specimen containers <NUM> from the carriers <NUM> after analysis is completed. Otherwise, a suitable offloading lane (not shown) may be provided elsewhere on the track <NUM>.

Again referring to <FIG>, a robot <NUM> may be provided at the loading area <NUM> and may be configured to grasp the specimen containers <NUM> from the one or more racks <NUM> and load the specimen containers <NUM> onto the carriers <NUM>, such as on an input lane or other location of the track <NUM>. The robot <NUM> may include one or more (e.g., at least two) robot arms or components capable of X and Z, Y and Z, X, Y, and Z, r and theta, or r, theta, and Z motion. Robot <NUM> may be a gantry robot, an articulated arm robot, an R-theta robot, or other suitable robot type wherein the robot <NUM> may be equipped with robotic gripper fingers that may be sized to pick up and place the specimen containers <NUM>. In one or more embodiments, the specimens may be placed in the carrier <NUM> in a predefined rotational orientation such that the labels <NUM> are provided on a back side of the specimen container <NUM>, away from the front side that will be imaged (as shown in <FIG>, <FIG>, and <FIG>) so that the label <NUM> will not occlude a direct view of the specimen.

To obtain this orientation, the operator may install them in a defined rotational orientation in the rack <NUM>, or the robot <NUM> may pick up the specimen container <NUM> and scan the identification information 218i to determine the label location and then place the specimen container in the carrier <NUM> in the pre-defined orientation based upon knowing the fixed location of the barcode on the label <NUM>. Other means for properly placing the specimen container <NUM> with label <NUM> located to the back side may be used.

Upon being loaded onto track <NUM>, the specimen containers <NUM> carried by carriers <NUM> may progress to a centrifuge <NUM> (e.g., a device configured to carry out fractionation of the specimen <NUM>). Carriers <NUM> carrying specimen containers102 may be diverted to the centrifuge <NUM> by inflow lane or a suitable robot (not shown). After being centrifuged, the specimen containers <NUM> may exit on outflow lane, or otherwise be moved by the robot, and continue on the track <NUM>. In the depicted embodiment, the specimen container <NUM> in carrier <NUM> may next be transported to a characterization apparatus <NUM> to be further described herein.

Characterization apparatus <NUM> is configured to characterize, through the use of hyperspectral imaging, the specimen <NUM> contained in the specimen container <NUM>, and may also be adapted to characterize the specimen container <NUM>. Quantification of the specimen <NUM> may include determination of HSP, HSB, or even HTOT, and may include determination of location of LA, SB and/or SG, and/or BG. The characterization apparatus <NUM> including hyperspectral imaging may also be configured for determining a presence of an interferent, such as one or more of hemolysis (H), icterus (I), and/or lipemia (L) contained in a specimen <NUM>.

In some embodiments, quantification of one or more physical attributes of the specimen container <NUM> may take place at the characterization apparatus <NUM> such as determining HT, TC, or even cap color or cap height. Once the specimen <NUM> is characterized and has passed predefined pre-screening criteria, the specimen <NUM> may be forwarded to be analyzed in the one or more analyzers (e.g., first, second, and third analyzers <NUM>, <NUM>, and/or <NUM>).

Additionally, one or more remote stations <NUM> may be provided on the specimen testing apparatus <NUM> even though the remote station <NUM> is not directly linked to the track <NUM>. Remote station <NUM> may be used to test for certain constituents, such as a hemolysis level, or may be used for further processing, such as to lower a lipemia level through one or more additions, for example. Other testing or processing may be accomplished on remote station <NUM>. For example, another characterization apparatus <NUM> may be located at the remote station <NUM>. An independent robot <NUM> (shown dotted) may carry specimen containers <NUM> containing specimens <NUM> to the remote station <NUM> and return them after testing/processing. Optionally, the specimen containers <NUM> may be manually removed and returned. Furthermore, in some embodiments, additional stations (not shown) may be arranged around the track <NUM> at various desirable locations, such as additional characterization apparatus <NUM>, a de-capping station, or the like.

The specimen testing apparatus <NUM> may include sensors <NUM> at one or more suitable locations around the track <NUM>. Sensors <NUM> may be used to detect a location of specimen containers <NUM> along the track <NUM> by means of reading the identification information 218i (see <FIG>) placed on the specimen container <NUM>, or like information (not shown) provided on each carrier <NUM> and communicating with computer <NUM>. In some embodiments, a barcode or RFID chip may be provided on the carrier <NUM> to aid in the tracking operation, for example. Other means for tracking the location of the carriers <NUM> may be used, such as proximity sensors. All of the sensors <NUM> may interface with the computer <NUM> so that the location of each specimen container <NUM> may be known at all times.

Specimen testing apparatus <NUM> may be controlled by the computer <NUM>, which may be a microprocessor-based central processing unit (CPU), having a suitable memory and suitable conditioning electronics, drivers, and software for carrying out the various computations and for operating the various components. Computer <NUM> may be housed as part of, or separate from, the base <NUM>. The computer <NUM> may operate to control movement of the carriers <NUM> to and from the loading area <NUM>, motion about the track <NUM>, and motion to and from the centrifuge <NUM>, motion to and from the characterization apparatus <NUM>. Computer <NUM> may also control operation of the characterization apparatus <NUM>. Computer <NUM> or a separate computer may control operation of the centrifuge <NUM>, and motion to and from each analyzer <NUM>, <NUM>, and <NUM>. In some embodiments, a separate integrated computer may control operation of each analyzer <NUM>, <NUM>, <NUM>.

For all but the characterization apparatus <NUM>, the computer <NUM> may control the specimen testing apparatus <NUM> according to software, firmware, and/or hardware commands or circuits such as those used on the Dimension® clinical chemistry analyzer sold by Siemens Healthcare Diagnostics Inc. of Tarrytown, New York, and such control is typical to those skilled in the art of computer-based electromechanical control programming and will not be further described herein. However, other suitable systems for controlling the specimen testing apparatus <NUM> may be used. The control of the characterization apparatus <NUM> may also be provided by the computer <NUM>, but according to methods described herein.

Embodiments may be implemented using a computer interface module (CIM) <NUM> that allows the user to readily access a variety of status and control display screens. These screens may describe some or all aspects of a plurality of interrelated automated devices used for preparation and analysis of specimens <NUM>. The CIM <NUM> may be employed to provide information about the operational status of a plurality of interrelated automated devices, as well as information describing the location of any specimen <NUM> as well as a status of tests to be performed on, or being performed on, the specimen <NUM>. The CIM <NUM> may thus be adapted to facilitate interactions between an operator and the specimen testing apparatus <NUM>. The CIM <NUM> may include a display screen adapted to display a menu including icons, scroll bars, boxes, and buttons through which the operator may interface with the specimen testing apparatus <NUM>. The menu may comprise a number of function buttons programmed to display functional aspects of the specimen testing apparatus <NUM>.

Pre-screening the specimen <NUM> and quantification as described herein may ensure that the specimen <NUM> can be stopped from progressing to the one or more analyzers <NUM>, <NUM>, and <NUM> if there is insufficient amount of serum or plasma portion 212SP available to carry out the ordered tests. Advantageously also, the ability to accurately quantify the physical location of LA and SB or SG may minimize not only the possibility of aspirating air, but also minimize the possibility of aspirating either settled blood portion 212SB or gel separator <NUM> (if the gel separator <NUM> is present). Thus, clogging and contamination of the specimen aspirating pipette used to aspirate serum or plasma portion 212SP for the analyzers <NUM>, <NUM>, <NUM> or other station(s) may be avoided or minimized.

Now with reference to <FIG>, a description of a first embodiment of a characterization apparatus <NUM> including hyperspectral imaging is provided. Characterization apparatus <NUM> may be configured to automatically characterize and/or quantify the specimen <NUM> (e.g., the serum or plasma portion 212SP, the settled blood portion 212SB, or both) and/or may quantify geometrical features of the specimen container <NUM>. The information obtained by the characterization apparatus <NUM> may also allow for identification of H, I, and/or L, and/or N of the specimen <NUM>. Thus, using the characterization apparatus <NUM> may help avoid one or more of gripper crashes, pipette clogging, and air aspiration by the pipette, and identifying HILN such that valuable analyzer resources are not wasted and that confidence in the test results may be improved.

Now referring to <FIG>, <FIG>, a first embodiment of a characterization apparatus <NUM> including hyperspectral imaging is shown. Characterization apparatus <NUM> may include an imaging location <NUM> configured to receive a specimen container <NUM> containing a specimen <NUM>, a light source <NUM> configured to provide lighting of the imaging location <NUM>, and a hyperspectral image capture device <NUM> configured to generate and capture a spectrally-resolved image of a portion of the specimen container <NUM> and specimen <NUM> at the imaging location <NUM>.

The light source <NUM> may be embodied as a light panel assembly including switchable light elements. In some embodiments, the light source <NUM> comprises a broad band light source. For example, the light source <NUM> may comprise one or more light panel assemblies, which may include a plurality of light sources. The light sources may collectively emit and provide a broadband light spectrum or a multi-band spectrum, wherein in the multi-band case, all of the light emissions from the multi-bands may be illuminated simultaneously.

In some embodiments, the broad band light source may provide a broadband emission anywhere in the range of <NUM> to <NUM>. For example, in some embodiments, one or more white light sources having a light emission range between about <NUM> and <NUM> may be used. In others, a broadband light source emitting near infrared (NIR and/or mid-IR) may be used, for example, emitting spectral output in the range between about <NUM> and <NUM>,<NUM>, wherein NIR may be considered as a wavelength range of between about <NUM> to about <NUM>, and mid-IR may be considered as a wavelength range of about <NUM> to about <NUM>,<NUM> herein. In some embodiments, combinations of white light and NIR light emissions may be used. In some embodiments, a broadband spectral range of greater than <NUM> within the spectral range of <NUM> to <NUM>,<NUM> may be used.

Furthermore, in some embodiments, multi-bands having discrete wavelength ranges within the spectral range of <NUM> and <NUM>,<NUM> may be used. For example, the light source <NUM> may simultaneously illuminate at multiple different spectra within the range of <NUM> to <NUM>,<NUM>. For example, multiple different discrete lighting elements may be illuminated at a time. For example, lighting elements <NUM> may be different colored LEDs, such as red LEDs (R), green LEDs (G), and blue LEDs (B) that emit light spectra at different nominal wavelengths. The light source <NUM> may simultaneously emit red light at <NUM> +/- <NUM>, green at <NUM> +/-<NUM> and blue at <NUM> +/-<NUM>, for example. In particular, the light arrays <NUM>, 456R may include clusters of R, G & B LEDs as lighting elements <NUM> that may be repeatedly arranged along the height of the light arrays <NUM>, 456R, such as in order RGB, RGB, RGB, etc.. High power Oslon SSL model LEDs available from Osram Opto Semiconductors GmbH of Regensburg, Germany may be used, for example. Each of the different-colored LEDs may be illuminated at once. For example, each or the R, G, and B LEDs as lighting elements <NUM> may be turned on simultaneously to provide multi- band illumination from the light source <NUM> to illuminate the specimen container <NUM> containing specimen <NUM> during imaging thereof. It should be recognized that R, G, and B are only examples, and that other combinations of discrete light elements may be simultaneously illuminated, such as any combination of R, G, B, UV, white light, NIR (wavelength range of about <NUM> to about <NUM>,<NUM>), and/or mid IR (wavelength range of about <NUM>,<NUM> to <NUM>,<NUM>), and the like in the spectral range of <NUM> to <NUM>,<NUM>.

Thus, one or more embodiments of light panel assembly may include at least two switchable lighting elements having different emission spectra. In some embodiments, switchable R, G and B lighting elements are provided. In some embodiments, switchable R, G, B, and white lighting elements are provided. In yet other embodiments, switchable R, G, B, and UV lighting elements are provided. In yet other embodiments, switchable R, G, B, and NIR or mid-IR lighting elements are provided. Any combination of two or more of switchable R, G, B, white light, UV, NIR, and mid-IR lighting elements may be provided in the light panel assembly. For NIR, an LED having a wavelength of <NUM> +/- <NUM> may be used in some embodiments. In such embodiments, the combination of switchable lighting elements may be provided in equal amounts and approximately evenly spaced along the height of the light guide <NUM>.

Referring to <FIG>, the characterization apparatus <NUM> including hyperspectral imaging may include light source <NUM> as an active backdrop, as shown, i.e., that may be provided by the light panel assembly to provide back lighting. That is, the one or more light panel assemblies are located on a back side of the imaging location <NUM> opposite from the hyperspectral image capture device <NUM> and configured to provide back lighting of the specimen container <NUM> and specimen <NUM> at the imaging location <NUM>.

The light source <NUM> may be embodied as a light panel assembly as shown in <FIG>, wherein each light panel assembly may include a frame <NUM>, a light guide <NUM>, and light assembly <NUM> configured to emit light into the light guide <NUM> and provide light emission from a front surface <NUM> of the light source <NUM>. In the depicted embodiment, the light assembly <NUM> may emit light into the lateral edges L, R (e.g., the side edges) of the light guide <NUM>, as best shown in <FIG>. The light source <NUM> may further include a diffuser <NUM>, where one panel surface of the diffuser <NUM> may be the panel front surface <NUM>. Protective films may be used on or in conjunction with the diffuser <NUM>.

The frame <NUM> may be made of a rigid material, such as plastic, and may include suitable fastening structures, such as bores <NUM> that are adapted to be mounted onto fixed mounting rods 455R (<FIG>). Other suitable mounting features may be included for mounting the light source <NUM> in a fixed orientation to the imaging location <NUM>. Frame <NUM> may include a pocket <NUM> that may include an open front and top and a closed back surface 458B and bottom, as shown, and may be configured to receive and position the light assembly <NUM>, the light guide <NUM>, and the diffuser <NUM> (if used) therein. The light assembly <NUM>, light guide <NUM>, and diffuser <NUM> may be inserted into the pocket <NUM> from the top and secured in place with securement member <NUM>. Other means for securing the light assembly <NUM>, light guide <NUM>, and the diffuser <NUM> in the frame <NUM> may be used.

The light guide <NUM> may be made of any suitably transparent light guide material including light diffusing capability, such as provided by a plastic sheet including internal light diffusing particles or other means of internal light diffusion. One suitable material is Acrylite LED® EndLighten, a product available from Evonik Industries AG of Essen, Germany. The light guide <NUM> may be made of a sheet having a width of between about <NUM> and about <NUM>, a height of between about <NUM> and <NUM>, and a thickness of between about <NUM> and about <NUM>, for example. Other suitable sizes and thicknesses may be used.

In the depicted embodiment of <FIG>, the light guide <NUM> may function by guiding light emitted laterally into the lateral edges L, R thereof by light arrays <NUM>, 456R (LED strip modules) of the light assembly <NUM> through the bulk material of the light guide <NUM> and emitting light on the front face 454F and rear surface 454R of the light guide <NUM> due to light interactions with the light diffusing particles therein. In some embodiments, the rear surface 454R of the light guide <NUM> may include a highly-reflective material formed thereon to reflect or backscatter any light transmission passing towards the back surface 458B and direct it back into the bulk material of the light guide <NUM> so that it may then be emitted from the front face 454F. Optionally, a highly-reflective material may be provided on the back surface 458B of the frame <NUM>, or as an individual element between the back surface 458B and the light guide <NUM>. The highly-reflective material may be provided as a mirror or a white plastic element, or other plastic or glass element with a metallic coating of silver, gold, chrome, tin, or combinations, for example. The light emitted from the front face 454F may be radiated substantially uniformly across the entire surface of the light guide <NUM> and illuminates the specimen container <NUM> and specimen <NUM> at the imaging location <NUM>.

The light arrays <NUM>, 456R may be LED strip modules including a linear array of individual light source elements (e.g., light emitting diodes - LEDs) arranged linearly along the lateral edges L, R of the light guide <NUM>. The light arrays <NUM>, 456R each may include a plurality of LEDs, such as between about <NUM> and <NUM> LEDs, for example, that may be arranged on a circuit board with a connector 456C provided to allow electrical connection with and operation by, the computer <NUM>. The light arrays <NUM>, 456R may be provided along the respective sides of the pocket <NUM> and are configured such that the emitting portion of each (e.g., LEDs) are provided directly adjacent to the lateral edges L, R and even touching the lateral edges L, R, if possible.

The light arrays <NUM>, 456R may be switchable, i.e., may be rapidly switched on and off to provide illumination of the imaging location <NUM>. The switching of the lighting elements <NUM> may be accomplished by software operable on the computer <NUM> coupled with an appropriate power source and drivers. The switching of the lighting elements <NUM> may coincide with the image capture.

The optional diffuser <NUM> including diffusing properties and may be provided as a sheet of Acrylite® Satince available from EVONIK of Essen, Germany in some embodiments. The 0D010 DF colorless was found to work well. The diffuser <NUM> may be a sheet having height and width dimensions approximately the same as the light guide <NUM> and a thickness of between about <NUM> and about <NUM>, for example. Other dimensions may be used. The diffuser <NUM> may function by scattering light passing through it. The diffuser <NUM> and the light guide <NUM> may be provided in spaced relationship to one another with a slight gap formed there between. The gap may be, for example, between about <NUM> and about <NUM>, and about <NUM> in some embodiments. Other gaps may be used.

The characterization apparatus <NUM> may include a housing <NUM> (shown dotted) that may at least partially surround or cover the track <NUM> and the imaging location <NUM>. The housing <NUM> may be a box-like structure provided to minimize outside lighting variances. Housing <NUM> may include one or more doors (not shown) to allow the carriers <NUM> to enter into and/or exit from the housing <NUM>. In some embodiments, the ceiling may include an opening to allow a specimen container <NUM> to be loaded into a carrier <NUM> by a robot including a gripper adapted to grasp the specimen container <NUM>.

The hyperspectral image capture device <NUM> will now be described in detail with reference to <FIG>. Hyperspectral image capture device <NUM> may include a first lens <NUM>, which is shown as a convex lens, including optical properties (e.g., focal length) configured to focus an image of a front surface FS (see also <FIG>) of the specimen container <NUM> and specimen <NUM> onto a plane of a slit aperture <NUM>. The first lens <NUM> is shown as a single lens. However, other lens systems including combinations of more than one lens may be used to accomplish this function. Slit aperture <NUM> may be a slit-shaped opening in a wall structure having a wide dimension L as shown in <FIG> that is aligned with a length dimension of the specimen container <NUM>, and a narrow dimension W as shown in <FIG> aligned with a width dimension across the specimen container <NUM>, wherein the large dimension L is much greater than the narrow dimension W. By way of example, and not by limitation, the wide dimension L may be between about <NUM> and <NUM>, and the narrow dimension W may be between about <NUM> and <NUM>. Other slit dimensions may be used. The focal length of the first lens <NUM> may be between about <NUM> and <NUM>, for example. Other suitable focal lengths may be used. Thus, the light emerging from the slit aperture <NUM> is representative of only a small width region (an imaged region - IR) as shown in <FIG>. The size of the slit aperture <NUM> may be chosen so that it is sized to provide a suitable image of the regions of the specimen container <NUM> and specimen <NUM> of interest. For example, the width of the imaged region IR may be between about <NUM> and <NUM> and may be located at an approximate center of the width of the specimen container <NUM>. The dimension of the width region is less than the overall width of the specimen container <NUM>. Other suitable widths may be used. The length L of the imaged region IR may encompass only the serum or plasma portion 212SP in some embodiments, but in other embodiments may include some or all of the cap <NUM>, some or all of the region including air <NUM>, serum or plasma portion 212SP, and the settled blood portion 212SB. In other embodiments, the length L of the imaged region IR may include only the serum or plasma portion 212SP and the settled blood portion 212SB. The location of the serum or plasma portion 212SP may be determined by any suitable segmentation method.

Hyperspectral image capture device <NUM> may further include a lens system including a second lens <NUM> and a third lens <NUM>, each of which may be concave lenses, wherein the second lens <NUM> may be located on a first side and the third lens <NUM> may be located on a second side of a spectrally-resolving element <NUM>. The lens system including second lens <NUM> and third lens <NUM> is configured and operable to project an image of the plane of the slit aperture <NUM> onto a spectral image capture device <NUM>. Thus, the second lens <NUM> operates to focus the light passing through the slit aperture <NUM> onto the spectrally-resolving element <NUM>, and the third lens <NUM> focuses the dispersed spectral image onto the spectral image capture device <NUM>. Any suitable lens or lens system may be used to accomplish these functions.

The spectrally-resolving element <NUM> is a device that spectrally disperses the incident light received thereat into a broader light spectrum, i.e., where the wavelength components (e.g., color or other components) making up the light spectrum are separated (like a rainbow) into continuous or discrete wavelengths.

The focal length of the second lens <NUM> may be between about <NUM> and <NUM>, for example. The focal length of the third lens <NUM> may be between about <NUM> and <NUM>, for example. Other focal lengths may be used.

The spectrally-resolving element <NUM> may be, for example, a prism, a diffraction grating, a spatially-varying filter, such as a linear band pass filter, or the like. A prism is a glass or other transparent object in prism form, especially one that is triangular with refracting surfaces at an acute angle with each other and that separates broadband light (e.g., separates white light into a spectrum of colors). A diffraction grating is an optical component with a periodic structure, which splits and diffracts light into several beams travelling in different directions. The diffraction grating may be transmissive or reflective. Other suitable devices for separating the light passing from the slit aperture <NUM> into its spectral components along the narrow dimension W may be used.

In the case of the prism, a right-angled or other suitable prism design may be used. The prism may include NBK-<NUM> borosilicate glass, B270 crown glass, N-SF1 glass, or fused silica glass, or the like, for example. In the case of a spatially-varying filter, such as a linear band pass filter, certain regions of the filter will operate to transmit/pass only certain wavelength ranges. Thus, different physical regions of the filter may pass different wavelength ranges, so that the incident light on the filter is spatially resolved into numerous discrete wavelength bands which than are projected onto the spectral image capture device <NUM>.

The spectral image capture device <NUM> may be any suitable detector capable of detecting light in the spectral wavelength range of interest, i.e., that can detect the spectrally-dispersed light. For example, the spectral image capture device <NUM> may be a photodetector, a charged coupled device (CCD), an array of photodetectors, array of phototransistors, array of photodiodes, one or more CMOS sensors, or the like. The resolution should be sufficient to match a predefined output spectral resolution. For example, the spectral image capture device <NUM> may include a resolution sufficient to produce spectral image size of about <NUM> pixels in the Z direction (corresponding to the wide dimension L of the slit aperture <NUM>) and about <NUM> pixels in the Y direction (corresponding to the narrow dimension W of the slit aperture <NUM>), for example. Other pixel densities and ratios of sizes in the L to Y dimensions may be used. In particular, a length of the spectral image capture device <NUM> in a direction associated with the L dimension may be greater than a width of the spectral image capture device <NUM> in a direction associated with the Y dimension. Further, the spectral image capture device <NUM> may have a spectral resolution of at least <NUM> pixels per cm (<NUM> pixels per inch) in the Y dimension, and may have a spectral resolution of at least <NUM> pixels per cm (<NUM> pixels per inch) in the L dimension. The length of the spectral image capture device <NUM> may be between <NUM> and <NUM>, for example. Other sizes may be used.

The spectral image capture device <NUM> communicates the captured image data of the spectrally-resolved image to the computer <NUM>, which is configured and operable to process the data of the spectrally-resolved image received at the spectral image capture device <NUM> and determine at least one of: segmentation of the specimen and/or specimen container, and determination of a presence or absence of an interferent in the serum or plasma portion 212SP of the specimen <NUM> (e.g., a fractionated specimen).

An example of a spectral image capture device <NUM> is shown in <FIG> and may include a resolution of about <NUM> pixel units associated with the Z (height) dimension of the specimen container <NUM>, and a resolution of about <NUM> pixel units associated with the Y dimension, which is the spectral dimension. Other numbers of units are possible, depending upon the degree of in the measurement accuracy that is sought. Calibration of the vertical Z dimension may be accomplished depending on the optics and the amount of spread of the image in the Z direction. In any event, if needed, calibration will enable direct correlation between the vision system and the actual physical coordinate location along the specimen container <NUM> in the Z dimension.

Hyperspectral image capture device <NUM> may include an enclosure <NUM> configured to house and support the various components thereof, such as the first lens, <NUM>, aperture assembly including slit aperture <NUM>, second lens <NUM>, third lens <NUM>, spectrally-resolving element <NUM>, and spectral image capture device <NUM> in a suitable orientation so that an image of the specimen container <NUM> and specimen <NUM> is from a single lateral viewpoint, as shown. <FIG> includes a coordinate break <NUM> to allow the third lens <NUM> and spectral image capture device <NUM> to be shown in parallel to the other components for clarity.

The imaging location <NUM> is a location in the housing <NUM> including an expected location of the specimen container <NUM>. In some embodiments, the specimen container <NUM> may be placed at or stopped at the imaging location <NUM>, such as by stopping the carrier <NUM> on the track <NUM> or otherwise placing the specimen container at the imaging location <NUM> by a robot (or even manually), so that it is approximately located in a center of the image window of the hyperspectral image capture device <NUM>.

<FIG> and <FIG> will be used to illustrate the processing of the spectral image data received at the spectral image capture device <NUM>. The image focused onto the spectral image capture device <NUM> is a spectrally-resolved image in the Y dimension, yet still correlated spatially to the height of the specimen container <NUM> in the Z dimension (see <FIG>). The imaged region IR (<FIG>), may be a small portion of the width of the specimen container <NUM>, such as approximately at or near the center thereof, but which is less than the full width of the specimen container <NUM>. The imaged region IR extends along at least a portion of the height (Z dimension) of the specimen container <NUM>, and even extending slightly above the cap <NUM> and/or below the bottom of the tube <NUM> in some embodiments. At each vertical location, correlated with each row of image units <NUM> along the Z dimension, the image is spectrally resolved in the Y dimension into the range of spectral interest. Thus, some or all of the spectral units <NUM> in the Y dimension each receive thereat a different small spectral portion of the overall range or ranges of spectra received at the spectral image capture device <NUM>. The spectral portion received at each spectral unit <NUM> depends on the resolution of the spectral image capture device <NUM> in the Y dimension and also on whether the light source is broadband or multi-band. The spectral resolution may be between about <NUM> pixel units and <NUM> pixel units over the width in the Y dimension and/or over the height in the Z dimension, for example. Other spectral resolutions may be used.

The spectral range of interest may be the same as the emitted spectra of the light source <NUM> in some embodiments, and therefore may be between about <NUM> and <NUM>,<NUM>. However, in some embodiments where white light is used, the emitted spectral range and thus the spectral range of interest may be between about <NUM> and <NUM>. The extent of the Z dimension used for computation depends on the results of segmentation. The segmentation process may be accomplished using the output of the hyperspectral image capture device <NUM>, or optionally by some other segmentation method. For example, in some embodiments, the segmentation may be performed using another method, such as is taught in <CIT>, for example. Once segmentation is completed, the extent of the Z dimension used for computation can be determined. In particular, the data associated with regions other that the serum or plasma portion 212SP in the Z dimension may be ignored for the interferent determination (e.g., for determining HILN). Thus, the portion of the imaged region IR above and below the serum or plasma portion 212SP may be unused.

At each vertical Z location associated with image units <NUM>, spectral data from spectral units <NUM> over the resolved spectral range received by the spectral image capture device <NUM> may be provided to the computer <NUM>, wherein each spectral unit <NUM> along the Y dimension (width) of the spectral image capture device <NUM> that receives light corresponds to a small portion of the spectral range received at the spectral image capture device <NUM>.

A representative spectral plot for various materials from the characterization apparatus <NUM> including hyperspectral image capture device <NUM> and back lighting with light source <NUM> is shown in <FIG>. The plot includes representative approximated blood spectral data signatures <NUM> for settled blood portion 212SB, gel separator spectral data <NUM>, normal spectral data <NUM> if a serum or plasma portion 212SP that is normal N, air spectral data <NUM>, and cap spectral data <NUM>. Thus, for each of the image units <NUM> in the Z dimension a representative intensity data plot of normalized intensity I (norm) versus each wavelength λ over the resolved spectral range may be obtained. Thus, for each image, a large amount of spectral information is captured. The captured image data represents intensity I (norm) measured by the spectral image capture device <NUM> at each wavelength λ, which then may be normalized to <NUM>. This data provides intensity responses over the wavelength spectra of interest for the material that is located at that associated Z dimension.

A similar data set may be generated for each respective vertical row of the image units <NUM>. Thus, a plurality of subsets of data, which may be in the form of a data matrix, may be obtained of intensity versus wavelength λ for some or all of the image units <NUM>. As discussed above, each subset of data may contain a spectral signature (e.g., data like in plot of <FIG>) wherein each is indicative of a particular class of material associated with that vertical Z location. Thus, processing the data with computer <NUM> to identify the representative signature may, in one or more embodiments, be used to provide segmentation. Segmentation as defined herein is the use of the data of the spectrally-resolved image to determine the respective classes of material of at least the specimen <NUM>, and possibly also the specimen container <NUM> along the vertical Z dimension.

The segmentation of the specimen container <NUM> and/or the specimen <NUM> may be accomplished by analyzing the various spectral signatures as a function of the Z dimension by any suitable analysis method. For example, the classification can be based, in one embodiment, on a set of expert-based rules. The expert-based rules may be based upon wavelength filters to process and examine only a certain frequency range or ranges, threshold limits, extrema (e.g., minima and/or maxima), or ranges within those frequency range or ranges that are indicative of certain material spectral signatures. One or more slopes or other characteristics may also be used as an expert rule.

For example, during a segmentation analysis, the imaged region IR in the wide Z dimension may be chosen to have a length L sufficient to encompass a full height of a tallest specimen container <NUM> that is expected to be received at the imaging location <NUM>. Referring again to <FIG>, the spectral signatures may be analyzed along image units <NUM>, from the top down. The cap <NUM> would first be encountered, which could be recognized by its very low transmittance. For example, the spectral signature of cap spectral data <NUM> of the cap <NUM> may be analyzed at <NUM> and if the intensity is less than an intensity threshold <NUM>, then cap <NUM> would be determined by the expert-based rule. Moving downward in the captured spectral data along image units <NUM>, the next element encountered would be air (air as seen through the specimen container <NUM>) including air spectral signature of air spectral data <NUM> that is representative of the class of air <NUM>, which may be discriminated by comparing the signature at <NUM> against an air threshold minimum <NUM>, for example. If above the air threshold minimum <NUM>, then that image unit <NUM> would be determined to be air <NUM>. Next, the liquid-air interface LA would be expected. Thus, a change to the associated spectral signature as shown in <FIG> or <FIG> illustrating various HILN conditions may be present. Normality N may be determined by identifying the spectral signature of normal spectral data <NUM> representative of being normal N, by using any form of expert rule. For example, dual liquid thresholds could be used at <NUM> and <NUM>, for example. Thus, a determination of normality N can be made for the particular image unit <NUM> in the vertical Z direction if a maximum liquid threshold <NUM> was not exceeded at <NUM>, while also a minimum threshold <NUM> is exceeded at <NUM>.

Other suitable expert rules may be used, including examining the location of local extrema (minima and/or maxima) in the spectrum or one or more slopes of the spectral signature at one or more wavelengths or between wavelengths. Next expected is either gel separator <NUM> or settled blood portion 212SB. Again expert rules can be used to determine these classes, as well. For example, an abrupt change in signature can be expected going from serum or plasma portion 212SP to settled blood portion 212SB, because of the very low transmittance of the settled blood portion 212SB. Thus, one or more thresholds on either slope and/or Intensity I (norm) may be used. The bottom most part of the specimen container <NUM> may be known based upon previous machine space calibration or machine space-image space calibration thereby knowing the exact location of the bottom register 122B (See <FIG>) of the receptacle receiving the specimen container <NUM> in the carrier <NUM>. Discrimination of gel separator <NUM> may be determined based on a slope or combination of slope or intensity threshold(s), for example. Other types of expert rules may be used for segmentation by analyzing the spectral data at each image unit <NUM> and subset thereof.

Other means for segmentation using the spectral image data may include using a model-based approach, wherein the various spectral data for some or all of the respective image locations in image units <NUM> and spectral units <NUM> may be feed as an input to a model, which then produces segmentation thereof. For example, in one embodiment, the segmentation may be carried out using an appropriately trained neural network as described herein.

Another embodiment of characterization apparatus <NUM> including a light source <NUM> configured to provide back lighting and hyperspectral image capture device <NUM> is shown and described with reference to <FIG>. Characterization apparatus <NUM> is similar to the embodiment of <FIG> and <FIG>, except that the spectrally-resolving element <NUM> is a diffraction grating and is embodied in a reflective configuration whereas the <FIG> embodiment is embodied as a transmissive configuration. The processing of the image data received at the spectral image capture device <NUM> may be the same as described herein. In this case where the spectrally-resolving element <NUM> is a diffraction grating, wherein separation into spectral components is by means of diffraction.

The light source <NUM> may be constructed as previously described, and the LEDs or other suitable lighting elements <NUM> arranged along the height of the light guide <NUM> may be broadband light sources such as white light LEDs or other white light emitting elements, or multi-band light elements (e.g., R, G, B). The slit aperture <NUM> may be as previously described.

Another embodiment of characterization apparatus 830A similar to characterization apparatus <NUM> is illustrated in <FIG>. Characterization apparatus 830A includes multiple ones of light sources 450A, 450B configured to provide illumination by front lighting of the imaging location <NUM>, and a hyperspectral image capture device <NUM> as previously described. The processing of the image data received at the spectral image capture device <NUM> may be as described herein. The spectral content for some of the components may be slightly different for the front side illumination case. However, processing may be accomplished by expert-based rules, or model-based methods configured to recognize or test for certain features of the spectral data. In an alternate embodiment, another light source 450C may be included in the characterizing apparatus 830A so that transmissive imaging may be provided in combination with front lighted imaging.

<FIG> illustrates yet another embodiment of a characterization apparatus 830B including hyperspectral imaging that may include multiple ones of the hyperspectral image capture devices 430A-C and multiple ones of the light source 450A, 450B, 450C arranged around the imaging location <NUM>. Three hyperspectral image capture devices 430A-430C are shown equally spaced, but more than three could be used. This configuration of characterization apparatus 830B maximizes the possibility that if the label <NUM> may be located on the front side in one viewpoint so as to occlude view of the serum or plasma portion 212SP, there may be an open viewing window to image the serum or plasma portion 212SP on at least one other lateral viewpoint, where the serum or plasma portion 212SP is fully visible or occluded only on a backside. Each hyperspectral image capture device 430A-C may include the configuration taught in any of the embodiments described herein, such as shown in <FIG>, <FIG>, and <FIG>.

Any of the characterization apparatus described herein (e.g., characterization apparatus <NUM>, <NUM>, 830A, 830B) may be used to determine an interferent, such as HIL, in the serum or plasma portion 212SP, following the segmentation process to determine the vertical location of the serum or plasma portion 212SP of the specimen <NUM>. As shown in <FIG>, which represents a schematic diagram <NUM> of the functional components of the characterization apparatus <NUM>, <NUM>, 830A, 830B, in block <NUM>, a specimen container <NUM> containing the specimen <NUM> (e.g., fractionated specimen) is provided at the imaging location <NUM>. The specimen container <NUM> containing the specimen <NUM> may be supported in an upright orientation in the carrier <NUM> as shown in <FIG>, for example. The carrier <NUM> may be configured to travel on a track <NUM> of a specimen testing apparatus <NUM> as shown in <FIG>.

In block <NUM>, a spectrally-resolved image of a portion (e.g., the imaged region IR - see <FIG>) of the specimen container <NUM> containing specimen <NUM> is captured by the spectral image capture device <NUM>. Optionally, in block 902A a non-spectral image of the specimen container <NUM> containing specimen <NUM> may be captured in addition to the spectrally-resolved image in <NUM>. The image data from either one of these can be used to accomplish segmentation of the specimen <NUM> and/or specimen container <NUM>, or both in segmentation block <NUM>. In particular, the goal of the segmentation in segmentation block <NUM> is to determine the vertical location of the serum or plasma portion 212SP in block <NUM>. In particular, the segmentation in segmentation block <NUM> may determine, as an optional output, the tube-cap interface TC in block <NUM>. This may be used to determine a height of the specimen container <NUM>. Segmentation in segmentation block <NUM> may also determine as an output, the liquid-air interface LA in block <NUM>, and may determine the serum-blood interface SB or the serum-gel interface SG in block <NUM>. Once segmentation in segmentation block <NUM> is completed, the region of interest can be isolated in block <NUM>, which may be to isolate any further analysis to only the serum or plasma portion 212SP, as the upper and lower boundaries thereof are now known.

In block <NUM>, the presence or absence of an interferent can be determined based on analyzing the spectrally-resolved image data obtained in block <NUM>. In particular, the interferent may be one or more of H, I, and L. Now referring to <FIG>, example spectral signatures of two levels of each of H, I, L and of N in the serum or plasma portion 212SP are provided. For example, high lipemic spectral data <NUM> indicative of a high level of lipemia L having relatively-low level of transmittance, and low lipemic spectral data <NUM> indicating a relatively-lower level of lipemia L (shown dotted) and having a relatively-higher level of transmittance are shown in <FIG>. Likewise, high icteric spectral data <NUM> indicating a relatively-high level of icterus I having a relatively-low level of transmittance, and low icteric spectral data <NUM> indicating a relatively-lower level of icterus I (shown dotted) and having a relatively-higher level of transmittance are shown in <FIG>. Furthermore, low hemolytic spectral data <NUM> indicating a relatively-high level of hemolysis H and having relatively-low level of transmittance, and high hemolytic spectral data <NUM> indicating a relatively-lower level of hemolysis H (shown dotted) and having a relatively-higher level of transmittance are shown in <FIG>. The spectral plot of the normal spectral data <NUM> of a specimen <NUM> that is normal N is also shown in <FIG>.

Some of the spectral signatures can be may be identified using expert rules, as described above, such as whether the specimen <NUM> is normal N. Likewise, the presence of lipemia L may be determined using expert rules. For example, for some levels of lipemia one or more thresholds based on Intensity I (norm) and/or slope(s) and/or local extrema (minima, maxima) at one or more wavelengths λ may be used.

In other embodiments, the determination of H, I, and/or L in block <NUM> may be accomplished by analyzing the spectral signatures by means of a suitable machine learning method. The machine learning method may include a training phase where a sufficient number of annotated samples (e.g., hundreds or even thousands) of different configurations and levels of H, I, and L are evaluated in the characterization apparatus <NUM>, <NUM>, 830A, 830A and are provided to form the classifier.

The annotation may involve graphically outlining various regions in a multitude of examples of specimen containers <NUM> having various specimen conditions, which is provided in a learning phase to form the classifier, along with the spectral information obtained from testing the example specimens. The classifier may be trained by annotating specimen conditions such as locations of air <NUM>, areas occluded by label <NUM>, locations of serum or plasma portion 212SP, locations of settled blood portion 212SB, locations of gel separator <NUM> (if included), and information about the type and level of interferent present, such as H, I, L and N. The number of levels of example specimens containing HIL may be provided in as many increments as is wanted from the classifier as an output. Thus, examples including different configurations of label <NUM> on the back side or even on the front side, different levels of serum or plasma portion 212SP, different levels of settled blood portion 212SB, different locations of gel separator <NUM>, and different index levels of HIL may be provided as training inputs along with the spectral matrices for each containing the Intensity I (norm) versus wavelength information as a function of the Z dimension obtained by testing in the characterization apparatus <NUM>, <NUM>, 830A, 830B. Areas of the holder <NUM> may be ignored, as well as areas having barcode provided thereon.

For example, the machine learning analysis may include a neural network based approach wherein the data of the spectrally-resolved portion of the specimen container <NUM> comprising the serum or plasma portion 212SP determined by the previous segmentation in <NUM> may be provided as a data matrix input to the neural network to classify the type of interferent present, such as H, I, and/or L or if the specimen <NUM> is normal N. The neural network may also output an index (a level) associated with one or more of H, I, and/or L. The input to the neural network may comprise intensity or transmittance information as a function of wavelength λ for each of the units <NUM> associated with the serum or plasma portion 212SP input as a data matrix for each specimen <NUM> tested.

In each of the characterization apparatus <NUM>, <NUM>, 830A, 830B described herein, the image capture may be triggered and captured responsive to a triggering signal sent by computer <NUM> and provided in communication lines when the computer <NUM> receives a signal that the carrier <NUM> is located at the imaging location <NUM>. A single image may be captured and that image is very informationally dense, including the spectral content information at each image unit <NUM>.

<FIG> illustrates a flowchart of a method of characterizing a specimen container <NUM> and/or a specimen <NUM>. The method <NUM> includes, in <NUM>, providing a specimen container (e.g., specimen container <NUM>, such as a capped blood collection tube) containing a specimen (e.g., specimen <NUM>) at an imaging location (e.g., imaging location <NUM>). Imaging location <NUM> may be inside of a characterization apparatus <NUM>, <NUM>, 830A, 830B). The specimen container (e.g., specimen container <NUM>) may be placed at the imaging location (e.g., imaging location <NUM>) by being transported thereto on a track (e.g., track <NUM>), by being placed there by a robot (e.g., robot <NUM> or the like), or manually.

The method <NUM> includes, in <NUM>, providing a hyperspectral image capture device (e.g., hyperspectral image capture device <NUM>, <NUM>) that is configured to capture an image at the imaging location (e.g., imaging location <NUM>). The method <NUM> includes, in <NUM>, providing one or more light sources (e.g., light panel assemblies <NUM>, 450A, 450B) configured to provide illumination of the imaging location <NUM>. The illumination may be provided by back lighting as shown in <FIG> or as front lighting as shown in <FIG>, or combinations thereof. The illumination may be provided as broadband illumination, and may include one or more subranges in the range of between about <NUM> and <NUM>,<NUM>. In some embodiments, multiple sources may be used in combination (e.g., white light, NIR, and/or mid-IR). In one embodiment, the illumination may be provided by a white light source (e.g., <NUM> - <NUM>). Other broadband spectral lighting ranges spanning at least <NUM> may be used. Further multi-band light sources may be used.

The method <NUM> includes, in <NUM>, capturing the spectrally-resolved image of a portion (e.g., the imaged region - IR) of the specimen container <NUM> and specimen <NUM> with the hyperspectral image capture device, and, in <NUM>, processing the spectrally-resolved image to determine at least one of: segmentation of the specimen and/or the specimen container, and a presence or absence of an interferent, such as HIL. The imaged region IR may be a small region having a width in the Y dimension that is a small fraction of the overall width of the specimen container <NUM>. Moreover, the length L of the imaged region IR may encompass at least the serum and plasma portion 212SP and at least some of the settled blood portion.

While the characterization apparatus <NUM> has been shown in <FIG> as being located such that the pre-screening characterization is performed immediately after centrifugation, it may be advantageous to include this pre-screening for HILN using the characterization apparatus (e.g., characterization apparatus <NUM>, <NUM>, 830A, 830B) directly on an analyzer (e.g., analyzer <NUM>, <NUM>, and/or <NUM>), or elsewhere in the specimen testing apparatus <NUM>. Furthermore, in some embodiments, the centrifugation may be performed prior to loading the racks <NUM> into the loading area <NUM>, so that the characterization apparatus <NUM>, <NUM>, 830A, 830B may be located at the loading area <NUM> and the pre-screening can be carried out as soon a specimen container <NUM> is loaded into a carrier <NUM>.

Claim 1:
A characterization apparatus (<NUM>), comprising:
an imaging location (<NUM>) configured to receive a specimen container (<NUM>) containing a specimen (<NUM>), wherein the specimen includes a settled blood portion and a serum or plasma portion;
a light source (<NUM>) configured to provide lighting of the imaging location (<NUM>);
a hyperspectral image capture device (<NUM>) comprising a spectrally-resolving element (<NUM>) and a spectral image capture device (<NUM>), the hyperspectral image capture device (<NUM>) configured to generate and capture a spectrally-resolved image of a portion of the specimen container (<NUM>) and specimen (<NUM>) at the spectral image capture device (<NUM>); and
a computer (<NUM>) configured and operable to process the spectrally-resolved image received at the spectral image capture device (<NUM>) and determine
a presence or absence of an interferent, the interferent being hemolysis, icterus and/or lipemia and at least one of:
segmentation of the specimen (<NUM>), or
segmentation of the specimen container (<NUM>) and the specimen (<NUM>).