Patent Publication Number: US-2021187502-A1

Title: Rapid assessment device for radiation exposure

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application claims priority to U.S. Provisional Application No. 62/584,199, filed on Nov. 10, 2017, and U.S. Provisional Application No. 62/584,204, filed on Nov. 10, 2017, and U.S. Provisional Application No. 62/584,208, filed on Nov. 10, 2017, which applications are hereby incorporated herein by reference in their entireties. 
    
    
     TECHNICAL FIELD 
     The present disclosure relates generally to a method and apparatus analyze components in a sample and in particular to determining an individual&#39;s level of exposure to radiation. 
     BACKGROUND 
     The ability to determine an individual&#39;s level of exposure to ionizing radiation (bio-dosimetry) due to a nuclear event is critical in the performing of triaging and determining the appropriate medical treatment of the exposed individuals. The following is a review of the current art for bio-dosimetry. 
     Molecular responses to ionizing radiation exposure fall in three broad categories according to the type of biomolecule chosen for analysis. These responses involve DNA, RNA, and protein. 
     Of these three, DNA responses occur first in the cell, i.e. DNA damage by radiation occurs instantly upon exposure. However, repair of the damage can take hours or even days to weeks, depending on the cell type and the nature of the damage. DNA damage occurs in several broad categories, including chromosomal breaks and rearrangements that are visible through the microscope, and point mutations including small (submicroscopic) deletions/insertions/rearrangements. Detection of chromosomal events is slow because the steps needed for their identification typically require cell culture followed by time-consuming analysis involving trained observers. The turn-around time is typically several days. Some chromosomal changes can be observed in cells that have not been cultured to metaphase, but these approaches require hybridization technologies followed by manual scoring involving trained observers. 
     Another type of cytogenetic response, that does not require cell culture, is the evaluation of micronuclei in erythrocytes. Micronuclei are formed by chromosomal fragments and by disruption of the mitotic spindle apparatus. Micronuclei are rare events and, in humans, erythrocytes bearing micronuclei are very efficiently filtered out by the spleen. In individuals who have had their spleen removed, micronucleated erythrocytes are not filtered out, and because the micronuclei are readily visible they can be efficiently counted by manual and by flow cytometric means. However, only about one adult per 1000 has been splenectomized, effectively negating this assay from broad-based population monitoring. 
     Another flow-cytometry-based approach might involve the analysis of rearranged chromosomes, e.g. dicentrics and/or translocations. In this approach, chromosomes that have undergone radiation-induced rearrangements would be detected and enumerated. Rearranged chromosomes would appear to be bi-colored because they would have been hybridized with whole chromosome paints in a manner that is analogous to in situ hybridization. However, for such flow-based analyses the chromosomes would need to remain in solution, meaning that hybridizations would also have to occur in solution. These chemistries have not been widely adapted due to technical difficulties related to the separation of unbound probe from the chromosomes. 
     Another approach to biological dosimetry involves protein analyses. Of the three biological response systems (DNA, RNA, protein), protein is the least well-characterized. There are several reasons why proteins may not be the subject of much research effort. First, the number of proteins is very large, perhaps an order of magnitude greater than the number of genes due to alternative splicing of mRNA and post-translational modification such as phosphorylation. Second, proteins often occur in complexes, sometimes making it difficult to enumerate quantities of specific peptide sequences. Third, proteins are all-too-readily denatured, which can interfere with common analysis methods such as enzyme-linked immunosorbent assay (ELISA). Finally, efficient quantitative analysis of proteins requires the use of monoclonal antibodies, which are not available for every protein, not to mention the vast array of protein variants that result from alternative splicing of mRNA and post-translational modification. Numerous changes to protein occur following irradiation. Unfortunately, the analytical techniques available for protein analysis lag substantially behind those for nucleic acid. To date, there are no systems using proteins that offer hope of performing meaningful biological dosimetry. 
     The third category of biological response to radiation is RNA. Specific RNA sequences are known to respond to ionizing radiation. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
         FIG. 1  is a perspective view of an example bio-dosimetry device. 
         FIG. 2  is a perspective view of the example bio-dosimetry device of  FIG. 1  in an open position as well as an example cartridge for performing analysis being received into bio-dosimetry device. 
         FIG. 3  is an exploded view of the cartridge shown in  FIG. 2 . 
         FIG. 4  is a block diagram of an example bio-dosimetry system including the bio-dosimetry device shown in  FIGS. 1 and 2  and the cartridge shown in  FIGS. 2 and 3 . 
         FIG. 5  is a perspective view of an example blister for injecting fluid into the cartridge at points in a processing flow. 
         FIG. 6A  is a top view of an example lysing region with a cover removed. 
         FIG. 6B  is a perspective view of an example lysing array with the cover removed. 
         FIG. 6C  is a side view of the example lysing array of  FIG. 6B  showing the cover and including a piezo electric layer. 
         FIG. 6D  is a side view of an example lysing array of  FIG. 6B  showing the cover and including a piezo electric element. 
         FIG. 6E  is a top view of a section of the example lysing array of  FIG. 6B  with the cover removed. 
         FIG. 6F  is a side view of the example the section of the lysing region of  FIG. 6E . 
         FIG. 7A  is a top view of an example binding region. 
         FIG. 7B  is a perspective view of a section of the stationary phase in the binding region of  FIG. 7A . 
         FIG. 7C  is a top view of a section of the stationary phase in the binding region of  FIG. 7A . 
         FIG. 7D  is a side view of a section of the stationary phase in the binding region of  FIG. 7A . 
         FIG. 8A  is a top view of an example mixing region. 
         FIG. 8B  is a perspective view of a portion of the mixing region of  FIG. 8A . 
         FIG. 9  is a block diagram of an example PCR region. 
         FIG. 10  is a top view of an example pre-PCR zone. 
         FIG. 11  is side view of the example pre-PCR zone of  FIG. 10  along an axis WW. 
         FIG. 12A  is a top view of an example PCR zone. 
         FIG. 12B  is a top view of the example PCR zone of  FIG. 12A  with the heating elements removed. 
         FIG. 12C  is a top view of a section of the PCR zone of  FIG. 12A . 
       
         
       
         FIG. 12D  is a side view along cross-section X-X of the section of the example PCR zone of  FIG. 12C . 
         FIG. 12E  is a side view along cross-section Y-Y of the section of the example PCR zone of  FIG. 12C . 
         FIG. 12F  is a perspective view of the section of the example PCR zone of  FIG. 12C . 
         FIG. 13  is an illustration of an example arrangement of a cartridge  14 . 
         FIG. 14  is a diagram of an example process for performing bio-dosimetry. 
         FIG. 15  is a diagram of an example process for lysing a sample. 
         FIG. 16  is a diagram of an example process for binding components of the sample. 
         FIG. 17  is a diagram of an example process for converting components of the sample to material for analysis. 
         FIG. 18  is a diagram of an example process for performing PCR on the material for analysis. 
         FIGS. 19A-19E  are graphs illustrating example responses of a respective genes to radiation. 
         FIG. 19F  is a graph comparing example in vivo and ex vivo gene responses to radiation. 
         FIG. 20A-20C  illustrate example comparisons of modeling results with ex vivo human data respectively for 12 hours, 24 hours and 48 hours post exposure. 
         FIG. 20D  illustrates an example receiver operating characteristic (ROC) for modeling exposure to radiation at 12 hours post exposure. 
         FIG. 21  illustrates example graphs of fluorescence v. cycles from PCR analysis of blood samples with varying degrees of lysing. 
         FIGS. 22A, 22B, 22C  illustrate an example fluorescent image from a PCR zone. 
         FIG. 23  illustrates an example amplification of two targeted genes with a PCR zone. 
     
    
    
     DETAILED DESCRIPTION 
     RNA levels can be quantified quickly, easily, and inexpensively. Cell culture is not required, and every tissue contains RNA, so the analyses are not limited to a particular cell type. Significantly, RNA is readily reverse-transcribed into cDNA which can be amplified via the polymerase chain reaction (PCR) and quantified by real-time PCR. This means that very small amounts of tissue can be analyzed rapidly, efficiently, and quantitatively. Furthermore, RNA analyses take advantage of the very rapid response to ionizing radiation. Further, specific sequences of RNA undergo increased expression within an hour of exposure, while other genes continue to have elevated expression up to 24 hours, providing a flexible range of options for performing bio-dosimetry. 
     A portable bio-dosimetry system that can perform rapid and reliable testing of radiation exposure is needed. Disclosed is a handheld field deployable real-time quantitative polymerase chain reaction (qPCR) based system that can assess exposure of individuals to ionizing radiation based on the levels of specific mRNA sequences present in the leukocytes from a drop of blood. The drop of blood may be obtained, for example, by a finger stick in a field or triage setting following a nuclear event. The system can predict whether an individual has received a dose of ionizing radiation greater than or less than a predetermined threshold value to allow field medical personnel to identify individuals in urgent need of medical treatment and to determine exposure levels to aid in medical treatment. 
     The portable-dosimetry system includes sub-systems for receiving a blood sample from a person, lysing (breaking up of) blood cells from the blood sample into component parts including mRNA, binding the mRNA in a binding region for further processing while discarding waste from the lysing process, converting the bound mRNA to complimentary cDNA and performing quantitative polymerase chain reaction (qPCR) on the complimentary cDNA. The results of the qPCR analysis can provide an indication of exposure to radiation of the person. 
       FIGS. 1-4  illustrate an example bio-dosimetry system  10 . The bio-dosimetry system  10  includes a bio-dosimetry device  12  and a cartridge  14  for processing a blood sample to determine the level of exposure to radiation of a respective person. As described in detail below, the bio-dosimetry device  12  may be a reusable, portable device that may be configured to receive one disposable cartridge  14  at a time, each cartridge  14  processing a blood sample of a respective person. The cartridge  14  is intended to be a single-use cartridge, used to measure a single sample. The bio-dosimetry device  12 , performs a number of functions on the sample, including pumping, lysing, binding, conversion and PCR analysis. Based on these operations, the bio-dosimetry device  12  can determine a level of exposure to radiation of the blood sample. 
     As an example, the bio-dosimetry system  10 , along with the various sub-systems and related processes, will be described in relation to analyzing a blood sample for exposure to radiation. It is envisioned within the context of this disclosure that the bio-dosimetry system  10  and/or some of the sub-systems, can be used for other applications such as analyzing cancer cells, types of cells (phenotypes) such as with use of flow cytometry, RNA viruses, types of bacteria, etc. 
     The bio-dosimetry device  12  comprises a housing  16 , a socket  18  for receiving the cartridge  14  and a door  20  coupled to the housing  16 . The door  20  encloses the socket  18  in a closed position ( FIG. 1 ) and provides access to the socket  18  in an open position ( FIG. 2 ). The housing  16  includes a reservoir  22  for receiving a blood sample and a cap  24  for enclosing the sample within the reservoir  22  (e.g., to avoid contamination). and a handle  26  for carrying the bio-dosimetry device  12 . The housing  16  further may include a display  28  for displaying information such as measurement data to a user, and a user input device  30  for receiving input from the user. 
     The door  20  may be coupled to the housing  16  with a hinge. When the door  20  is the opened position the socket  18  can receive the cartridge  14 . Upon receiving the cartridge  14  in the socket  18 , the door  20  can be closed. As described below, closing the door  20  can couple components in the bio-dosimetry device  12  to the cartridge  14 . 
     The bio-dosimetry device  12  includes a battery  48  which provides electrical energy to the bio-dosimetry device  12 . The battery  48  may be a standard battery such as a lead-acid, nickel-cadmium, lithium-ion battery etc. The battery  48  enables the bio-dosimetry device  12  to be portable and used in locations without access to a power grid. 
     The bio-dosimetry device  12  further include a computer  50  including one or more processors  52  and one or more memories  54 , the memories  54  including instructions for programming the one or more processors  52 . The computer  50  executes instructions which can be used carry out the processes for preparing and analyzing the sample for exposure to radiation as described herein. The computer  50  is communicatively coupled to components of the bio-dosimetry device  12  including the display  28 , the user input device  30 , sensors  56 , an illumination system  58 , optical detector  60 , heating systems  62 , pumps  64 , blister actuators  66  and a piezo electric element  68 . 
     The display  28  may be, for example, a liquid crystal display (LCD) or organic light emitting diode display (OLED) and can be used for outputting data, for example measurement data, to a user. The display  28  may further be a touch-screen device and used as an input device by the user. 
     The user input device  30  includes one or more input elements such as buttons, turn knobs, touch-screens, a microphone, etc. that permit a user to input data and/or instructions to the bio-dosimetry device  12 . 
     The sensors  56  can include, for example, temperature sensors, pressure sensors and contact sensors. The sensors  56  can receive instructions from and provide data to the computer  50  to enable the computer  50  to control the processes for preparing and analyzing the samples as described below. 
     The illumination system  58  provides light for performing PCR. The illumination system  58  is communicatively coupled to the computer  50  and includes a light source. The light source optimally emits light in a narrow band of wavelengths to stimulate the fluorescent tag. The fluorescent tag, in response, emits light at a different wavelength. The band of wavelengths radiated by the light source to stimulate the fluorescent tag needs to be limited to a range that does not overlap the light emitted by the fluorescent tag. There are many possible fluorescent tags (markers) and the wavelength of the light source needs to be matched to the tags. The light sources should be as monochromatic (narrow banded) as possible, such as a laser or a narrow emission light emitting diode or other light source with a narrow notch filter blocking wavelengths outside of the narrow band needed to stimulate the fluorescent tags. 
     The illumination system  58  may further include electronic circuitry to turn the light source on and off, bias the light source in an operating condition, control or limit the power supplied to the light source, etc. As described below, the illumination system  58  can radiate light on the cartridge  14  during analysis of material to be analyzed, based on commands from the computer  50 . Material to be analyzed as used herein is material that has been prepared to be analyzed based on components of the blood sample. The material to be analyzed may include fluorescent markers. 
     The optical detector  60  is communicatively coupled to the computer  50 , and can be, for example, a camera, a charge-coupled device (CCD), a light sensitive, CMOS array, etc. The optical detector  60  may include a filter to block the excitation wavelength (from the light source) used to stimulate a fluorescent tag and allow the fluorescent tag emission through for detection. Following or during illumination of the material to be analyzed by the illumination system  58 , the optical detector  60  receives light emitted by the material to be analyzed. The optical detector  60  can provide data to the computer  50  based on the received light. In an example, the material to be analyzed includes fluorescent markers. Following illumination, the fluorescent markers radiate light at a wavelength based on the markers and the optical detector  60  detects the light. In some cases, the optical detector  60  may be used to detect different fluorescent tags emitting light at different wavelengths. In these cases, the optical detector  60  may receive respective images of the two fluorescent tags by applying different filters for each of the respective fluorescent tags. 
     In at least one example, each of the heating system(s)  62  of the bio-dosimetry device  12  includes a heating element such as a resistance heater and may include electronic circuitry to turn the heating element on and off, bias the heating element in an operating condition, control or limit the power to the heating element, etc. The heating system  62  may further include a sensor  56 , such as a temperature sensor, that can be used to detect a temperature of the heating element and provide feedback to the heating system  62  such that the heating system  62  can regulate the temperature of the heating element. As will be explained in greater detail below, the heating system(s)  62  may be used to heat a portion(s) of the cartridge  14  during processing of the blood sample. Further, the heating elements included in the heating system  62  may in some cases be contained in the bio-dosimetry device  12  and in other cases included as part of the cartridge  14 . 
     In at least one example, the pump(s)  64  of the bio-dosimetry device  12  may be electrically coupled to and controlled by the computer  50  such that the computer  50  can turn them on and off. As described in additional detail below, when the cartridge  14  is inserted into the bio-dosimetry device  12 , and the door  20  is closed, the pumps  64  may further be in fluid communication with the cartridge  14  such that the pumps  64  can move fluids, e.g., from the reservoir  22  to the cartridge  14  or within the cartridge  14  from one region to another. 
     The bio-dosimetry device  12  may further include one or more blister actuators  66 . Each blister actuator  66  may be communicatively coupled with the computer  50  and include a plunger. The blister actuators  66  may be arranged within the bio-dosimetry device  12  such that, when the cartridge  14  is inserted in the bio-dosimetry device  12  and the door  20  is closed, the blister actuator  66  is within a range of elements on the cartridge  14  such that the plunger, in an extended state, extends into and compresses the elements on the cartridge  14  and releases fluids into a processing flow on the cartridge  14 . 
     The bio-dosimetry device  12  may further include a piezo electric element  38 . The piezo electric element  68  may be communicatively coupled with the computer  50  such that it can receive commands, for example, to turn the piezo electric element  68  on and off or adjust a power level of the piezo electric element  68 . When the piezo electric element  68  is turned on, it may vibrate at a frequency, for example  20  kHz. When a cartridge  14  is inserted into the bio-dosimetry device  12  and the door  20  is closed, the piezo electric element  68  may be coupled, either directly (in physical contact), or indirectly (for example, via a small air space) with elements on the cartridge  14  and cause the elements to vibrate, transferring mechanical energy to the elements. 
     The cartridge  14  is a disposable cartridge that is intended to be used to analyze a single blood sample and is configured to be inserted into the bio-dosimetry device  12 . The cartridge  14  includes a cassette  32  and a substrate  34 . The cassette  32  is adapted to be inserted into the socket  18  of the bio-dosimetry device  12  and includes a recessed area  40  for receiving and supporting the substrate  34 . The substrate  34  has a first side  41  and a second side  42  and supports a plurality of sections configured to process and analyze a blood sample and determine a level of exposure to radiation. The cartridge  14  further includes a cover  44  having a first side  45  and a second side  46 . The second side  46  of the cover is attached to the first side  41  of the substrate  34  and encloses the processing regions formed in the substrate  34 . The substrate  34  and the cover  44  can be made of a material such as glass (e.g., Pyrex), silicon, a silicone-based material, sapphire, a polymer, or any transparent substrate. 
     The cartridge  14  contains a plurality of regions for processing and analyzing a blood sample to determine a level of exposure to radiation. An overview of the regions of the cartridge  14  will be presented in the next paragraph. Thereafter, the structure and features of different sections of the cartridge  14  will be described in additional detail. 
     The blood sample is pumped into a lysing region  90  on the cartridge  14 , by a micropump  82  or via a port  84 . In the lysing region  90 , the blood sample is broken into components parts including mRNA. After lysing, the blood sample including the mRNA, is transferred to the binding region  92 . In the binding region  92 , mRNA from the blood sample is bound (held). The remaining components are removed, by flushing. Thereafter, the bound mRNA in the binding region  92  is flushed with a material, which forms cDNA with the bound mRNA. The cDNA, formed in the binding region  92  is then transferred to a mixing region  94  where the cDNA is more evenly distributed within a reaction mixture. Next, the cDNA is transferred to a polymerase chain reaction (PCR) region  96  where the cDNA is analyzed for exposure to radiation of the mRNA with which the cDNA was previously formed (in the binding region  92 ). 
     The micropump  82  may be built into and/or on the substrate  34  and based on a micro-electro-mechanic systems (MEMS) technology. When the cartridge  14  is inserted in the bio-dosimetry device  12  and the door  20  is closed, an input to the micropump  82  may be fluidly coupled to the reservoir  22  in the bio-dosimetry device  12  such that the micropump  82  can pump the sample into the cartridge  14  (for example, into the lysing region  90 ). 
     The cartridge  14  may further include one or more ports  84 . The ports  84  may be openings formed in the cover  44  of the cartridge  14 . The ports  84  may be arranged to fluidly couple, for example, with an outlet of a pump  64  in the bio-dosimetry device  12  when the cartridge  14  is inserted in the bio-dosimetry device  12  and the door  20  is closed. For example, the outlet of the pump  64 , which may be a nozzle or a tube, may be inserted into the port  84  when the door  20  of the bio-dosimetry device  12  is closed, such that when the pump  64  is activated, the pump  64  can output a fluid into the port  84 . 
     To store and, at a proper time during the sample processing, inject liquid materials in the processing flow, the cartridge  14  may include one or more blisters  86  fluidly coupled to respective regions in the cartridge  14 . For example, one or more blisters  86   a  may be fluidly coupled to an inlet to the lysing region  90 . One or more blister  86   b  may be coupled to an inlet to the binding region  92  and one or more blister  86   c  may be coupled to an inlet to the mixing region  94 . The blisters  86  may be, for example, a flexible, polymer based, fluid container, that is formed, for example, on a first side  55  of the cover  44 . 
     Microchannels  85  may be used to fluidly couple regions and features of the cartridge  14 . For example, a microchannel  85   a  may couple one or more of the micropump  82 , one or more blisters  86   a  and one or more ports  84  to an inlet of the lysing region. A microchannel  85   b  may fluidly couple an outlet of the lysing region  90  and an outlet of one or more blisters  86   b  to an inlet of the binding region  92 . A microchannel  85   c  may couple an outlet of the mixing region  94  to an inlet of the PCR region  96 , etc. Each of the microchannels  85  may be formed in the substrate  34  and closed on a first side  41  of the substrate  34  by a second side  46  of the cover  44 . 
     An example blister  86  is shown in  FIG. 5 . The blister  86  may be a hollow, flexible container made of a polymer that forms a cavity for containing a fluid. The blister  86  may have a flat portion  100  for attaching to a surface, for example the first side  45  of the cover  44 . The blister  86  may further have a dome-shaped structure  102  arranged on a side of the flat portion and forming the cavity. The dome shaped structure  102  may be, for example, round, or in the form of an elongated rectangle or oval. The blister  86  may have an outlet  104  which may be fluidly coupled, to one of the processing regions on the cartridge  14 . The blister  86  is configured such that, when pressure is applied to the dome shaped structure  102 , fluid contained in the blister  86  is expelled through the outlet  104 . 
     A blister  86  may be actuated by a blister actuator  66  on the bio-dosimetry device  12 . The blister actuator  66  may be arranged in the bio-dosimetry device  12  such that when the cartridge  14  is inserted in the socket  18 , the blister actuator  66  is in a range to actuate the blister  86 . At an appropriate time during a processing cycle, the computer  50  in the bio-dosimetry device  12  may instruct the blister actuator  66  to extend a plunger toward and apply pressure to the dome-shaped structure  102 , and expel the liquid contained in the blister  86  into the processing region to which the blister  86  is fluidly coupled. A size of each blister  86  in the cartridge  14  can be selected based on a volume of liquid required for the processing step in which the blister  86  participates. 
     An example lysing region  90  is illustrated in  FIGS. 6A-6F . The lysing region lyses biological samples by applying ultrasonic stimulation. In an example, the sample to be lysed may be a drop of blood. In other examples, the sample may be any fluid with cells, for example waver with bacteria, cells in lymphatic fluid, cells in urine, etc. The lysing region  90  may also be used for other applications such as lysing microbes, animal and plant cells, fracturing bonds in DNA, RNA, protein aggregates and lipid aggregates. The lysing region  90  may be applied to prepare a sample for polymerase chain reaction (PCR) and rolling circle genetic amplification. 
     The lysing region  90  includes an inlet  130 , a distribution manifold  132 , a lysing array  134 , an outlet manifold  136 , and an outlet  138 . The lysing region  90  is configured to receive a sample via the inlet  130 , pass the sample through the distribution manifold  132 , lyse, by transfer of mechanical energy, the sample into component parts in the lysing array  134  and output the component parts through the outlet manifold  136  to the outlet  138 . Each of the inlet  130 , the distribution manifold  132 , the lysing array  134 , the outlet manifold  136  and the outlet  138  can be formed in the substrate  34  ( FIGS. 6C, 6D ). 
     The inlet  130  can receive the sample and/or other input material from one or more of the micropump  82 , a blister  86  or a port  84 , and deliver the sample or input material to the lysing array  134  via the distribution manifold  132 . The distribution manifold  132  may be triangular shaped, with an apex of the triangle connected a microchannel  85   a  delivering the sample and/or input material and a base of the triangle connected to the lysing array  134 , such that the distribution manifold  132  broadens from the microchannel  85   a  to the lysing array  134  to distribute the material across a side of the lysing array  134 . 
     The lysing array  134  may be square and with a typical length of the side in a range from 0.5 cm to 1.0 cm. Other shapes are possible. For example, the lysing array  134  may be rectangular with either a short side of the rectangle of a long side of the rectangle connecting to the distribution manifold. Further, the size of the lysing array  134  is scalable. Increasing the size of the lysing array  134  increases a volume of material that can be lysed in a period of time. A smaller lysing array  134 , can be used, for example, to lyse viruses or other small components. 
     The outlet manifold  136  and outlet  138  output material from the lysing array  134  to a microchannel  85   b.  The outlet manifold  136  may be triangular shaped with a base of the triangle connected to the lysing array  134  and an apex connected to the microchannel  85   b,  such that the material output from the lysing array  134  is directed into the microchannel  85   b.    
     The lysing array  134  includes a diaphragm  155  formed in the substrate  34 . Referring to  FIG. 6B , the diaphragm  155  has a first side  156 , a second side  157  and a thickness t 1 . The thickness t 1  can be in a range from 0.01 millimeters to 1 millimeter with a typical value of 0.1 millimeter. Referring to  FIG. 6C , the first side  156  of the diaphragm  155 , together with the substrate  34  and the cover  44 , define a cavity  160 . The cavity  160  contains the sample to be lysed during lysing. The cavity  160  may have a typical volume of four microliters and a typical volume range from one to 100 microliters. This range is not intended to be limiting. In addition to cavity volumes within this range, a lysing array  134  according to this disclosure can also have volumes greater than or less than this range. 
     A plurality of micropillars  162  extend outwardly from the first side  156  of the diaphragm  155  into the cavity  160 . Each of the micropillars  162  has a top end  164 . In an example, the micropillars  162  are arranged in columns and rows and may be evenly spaced. As an example, the lysing array  134  may include 40 rows and 40 columns of micropillars  162 , for a total of 1600 micropillars. The micropillars  162  may be formed, for example, by an embossing or etching process. 
     The lysing array  134  may include, coupled to the second side  157  of the diaphragm  155 , a piezo electric layer  166 . During lysing, the piezo electric layer  166  may be stimulated (i.e., an electric current may be applied). Based on the stimulation, the piezo electric layer  166  may apply an oscillating force to the diaphragm  155 , causing the diaphragm  155  to flex. At points towards a center of the lysing array  134 , the flexing of the diaphragm  155  may be substantially perpendicular to a plane of the diaphragm  155 . In an example, the flexing of the diaphragm can result in a displacement in a range from 1 to 20 microns at points towards the center of the lysing diaphragm. In other examples, the displacement can be larger, up to a range of 1 millimeter or more. The amount of displacement may vary with the frequency of stimulation as there is a relaxation time dependent on displacement. That is, a lower frequency of stimulation may result in a larger displacement. The flexing causes a lateral motion on the top ends  164  of the micropillars  162 . The lateral motion of the top ends  164  can create a liquid shock wave that can lyse the sample contained in the cavity  160 , breaking the sample into component parts. An energy level applied to the diaphragm can be controlled to allow for lysing, for example, of cell walls, without damaging internal organelles in the blood cells. 
     As an alternative to the piezo electric layer  166 , a piezo electric element  68  included in the bio-dosimetry device  12  can be used to stimulate the diaphragm of the lysing region  90 . This is illustrated in  FIG. 6D . In this case, when the cartridge  14  is inserted in the bio-dosimetry device  12  and the door  20  closed, the piezo electric element  68  is arranged to physically couple with the diaphragm  155  such that when the piezo electric element  68  is turned on, ultrasonic waves radiated from the piezo electric element  68  cause the diaphragm  155  to flex. 
       FIG. 6E  is a top view of a section of the lysing array  134 , with the cover  44  removed. The section shows two rows R 1  and R 2  and two columns C 1  and C 2 . The micropillars  162  may be cylindrically shaped, and have a diameter d. The diameter d may, for example, be in a range from 0.01 millimeters to 10 millimeters with a typical value of 0.075 millimeters. The micropillars  162  may be evenly spaced and have a spacing W 1  between adjacent micropillars  162  in a row (e.g., the row R 1 ) and evenly spaced and have a spacing W 2  between two micropillars  162  in a column (e.g., the column C 1 ). In a typical example, W 1  may be substantially equal to W 2 . The spacings W 1  and W 2  may be in a range from 0.015 millimeters to 10 millimeters and have a typical value of 0.075 millimeters which is effective for most cells. The ranges of W 1  and W 2  may be selected based on a particle size of the sample to be lysed and would typically increase as the particle size to be lysed increases. 
     In another example, the lysing array  134  may be applied to open or break apart protein chains and viruses. In this case, the spacings W 1 , W 2  and the diameter d may be in a range of tens of nanometers. 
       FIG. 6F  is a side view of the section of the lysing array  134  shown in  FIG. 6D . The micropillars  164  can have a height h 1 . The height h 1  can be in a range from 0.1 millimeters to 100 millimeters and can depend on the diameter d chosen for the micropillars  164  and the sample to be lysed. The height h 1  and diameter d are typically selected such that an aspect ratio h 1 /d is 10, with a range from 5 to 20. For example, in the case that the diameter d is selected to be 2 millimeters, the height h 1  can be selected to be 20 millimeters. 
     The diaphragm  155  has a thickness t 1 . As noted above, the thickness t 1  can be in the range 0.01 millimeters to 1 millimeter with a typical value of 0.1 millimeters. The thickness of the piezo electric layer  166  (when present) may be in a range from 0.1 millimeters to 10 millimeters. 
     Other shapes may be used for the cross-section of the micropillars  162 . For example, the cross-section may be rectangular or triangular, the micropillars  164  may be conical with a base of the cone located at the first side  156  of the diaphragm  155 , etc. 
       FIGS. 7A-7D  illustrate an example binding region  92 . The binding region  92  includes a microchannel  200  having an inlet  203  and an outlet  204 . During operation, material, for example, a lysed blood sample flows through the microchannel  200  in a direction  212 . 
     The microchannel  200  includes a stationary phase  202 . The stationary phase  202  includes a plurality of micropillars  220 . The micropillars  220  are formed in and extend outwardly from the substrate  34 . As shown in  FIGS. 7B and 7C , the micropillars  220  are arranged in rows, for example rows R 3  and R 4 . Each row is offset from a previous row such that the micropillars  220  in the row R 4  are arranged to correspond with an opening between micropillars  220  in the row R 3 . 
     As an example, as shown in  FIG. 7C , the micropillars  220  may have a cross-section having a hexagonal shape. The micropillar  220  has first, second, third, fourth, fifth and sixth sides  222 ,  224 ,  226 ,  228 ,  230  and  232 . First and fourth sides  222 ,  228  are parallel, and have a length li. Second and fifth sides  224 ,  230  are parallel and have a length l 2 . Third and sixth sides  226 ,  232  are parallel and have length l 3 . In an example, the micropillar  220  has a diameter d 1  and a spacing between the micropillars d 2 . The diameter d 1  may be in a range of two microns to 2,000 microns, with a typical value of 20 microns. The spacing may be in a range from one micron to 1,000 microns, with a typical value of 10 microns. The micropillars  220  have a height h 2 . The height h 2  may be in a range of 10 microns to 10 millimeters, with a typical height of one millimeter. 
     In some cases, the micropillars  220  may be coated with a binding material to bind target components of the sample. For example, in the case of a blood sample containing mRNA, the micropillars  220  may be coated with oligo dT25 to bind the mRNA in the blood sample to the micropillars  220 . 
     The binding region  92  further may include a heating element  210  having two contacts  211  on opposite ends. The heating element  210  may be supported on the cover  44 . The heating element  210  may be formed of a resistive material such that, when an electrical voltage is applied across the two contacts  211 , a current will flow through the heating element  210  and generate heat. 
     Additionally or alternatively, during operation of the bio-dosimetry system  10 , a heating system  62  in the bio-dosimetry device  12  may be arranged such that when the cartridge  14  is inserted into the socket  18  and the door  20  closed, the heating system  62  is thermally coupled to (i.e., can heat) the binding region  92 . 
     The binding region  92  can be used to bind mRNA in a blood sample to the stationary phase  202 . This is described below as the process  1600  in reference to the  FIG. 16 . 
     The binding region  92  can further be used to convert the mRNA, bound in the stationary phase, to cDNA. This is described below as the process  1700  in reference to the  FIG. 17 . 
       FIGS. 8A and 8B  illustrate the mixing region  94 . The mixing region  94  includes a microchannel  240  having an inlet  241  and an outlet  242 . The microchannel  240  may be in the form of a meander having straight portions coupled at angles of, for example, 90° to one another. A length l 8  of a straight portion in the meander may have a typical value of 1 millimeter to 2 millimeters. The microchannel  240  is formed in the substrate  34  and has a width d 3 . The width d 3  can be in a range of 0.1 millimeters to 2 millimeters with a typical value of 0.5 millimeters. The depth U 4  of the microchannel  240  can be in a range from 0.1 millimeters to 1 millimeter with a typical value in a range from 0.1 to 0.2 millimeters. 
     The microchannel  240  can be used to remove gradients of cDNA in reaction mixture. The reaction mixture, containing the cDNA may enter the mixing region  94  at the inlet  241 , pass through the microchannel  240 , continue through the outlet  242 . As the reaction mixture containing the cDNA passes through the microchannel  240 , the changes of directions as the microchannel  240  passes through the  90  passing through 90° angles, may cause turbulence in the fluid flow, mixing the cDNA within the reaction mixture. Through the mixing process, the cDNA may be more evenly distributed, reducing or eliminating gradients in the cDNA within the reaction mixture. 
     The mixing region  94  may have other shapes or features. For example, the width d 3  of the microchannel  240  may be varied alternatively between a first width and a second width or the depth U 4  of the microchannel  240  varied alternatively between a first depth and a second depth to create turbulence in the reaction mixture flow. As another example, extensions or bumps may be formed, extending from a bottom of the microchannel  240  into the microchannel  240  to increase turbulence in the reaction mixture flow. 
       FIG. 9  is a diagram of an example PCR region  96 . The PCR region  96  receives material including cDNA from the mixing region  94 . The PCR region analyzes the cDNA for exposure to radiation of the mRNA from which the cDNA was generated. Analyzing the cDNA includes thermally cycling the material to be analyzed including the cDNA between two temperatures, as described below. The PCR region  96  receives the material via a microchannel  85   c.  The microchannel  85   c  couples the mixing region  94  ( FIG. 4 ) via four sub-microchannels  98   a,    98   b,    98   c,    98   d  to four respective PCR channels  250   a,    250   b,    250   c,    250   d.    
     A PCR region  96  including four PCR channels  250   a,    250   b,    250   c,    250   d  is only an example. The PCR region  96  may include any number of PCR channels  250 . 
     As shown in  FIG. 9 , the first PCR channel  250   a  includes the pre-PCR zone  252   a,  the blister  86   e  and the PCR zone  254   a,  the second PCR channel  250   b  includes the pre-PCR zone  252   b,  the blister  86   f  and the PCR zone  254 , the third PCR channel  250   c  includes the pre-PCR zone  252   c,  the blister  86   g  and the PCR zone  254   c  and the fourth PCR channel  250   d  includes the pre-PCR zone  252   d,  the blister  86   h  and the PCR zone  254   d.  The four PCR channels  250   a,    250   b,    250   c,    250   d  may be the same or similar, and will be described as an example PCR channel  250  including an example pre-PCR zone  252 , an example blister  86  and an example PCR zone  254  below. 
       FIGS. 10 and 11  illustrate an example pre-PCR zone  252 . The pre-PCR zone  252  heats material prior to beginning analysis in the PCR zone  254 . The pre-PCR zone  252  includes a microchannel  255  including an inlet  256  and an outlet  257 . Material enters the microchannel  255  at the inlet  256  and flows towards the outlet  257 . The outlet  257  may be coupled, for example, to a sub-microchannel  98  such as the sub-microchannel  98   a.  The outlet  257  may be coupled to the PCR zone  254 . 
     The microchannel  255  may have a width d 4  and a depth U 6 . Adjacent parallel sections of the microchannel  255  may be spaced away from each other by a distance d 5 . The width d 4  may be in a range from 0.1 millimeters to 2 millimeters with a typical value of 1 millimeter. The depth U 6  of the microchannel 255 may be in a range from 0.05 millimeters to 2 millimeters with a typical value of 1 millimeter. The spacing d 5  between adjacent sections of the microchannel  255  may have a range from 0.1 millimeters to 5 millimeters with a typical value of 2 millimeters. 
     The pre-PCR zone  252  further may further include a heating element  260  including two contacts  261 . on opposite ends. The heating element  260  may be supported on the cover  44 . The heating element  260  may be formed of a resistive material such that, when an electrical voltage is applied across the two contacts  261 , a current will flow through the heating element  260  and generate heat. 
     Additionally or alternatively to including the heating element  260 , a heating system  62  in the bio-dosimetry device  12  may be arranged such that when the cartridge  14  is inserted into the socket  18  and the door  20  closed, the heating system  62  thermally couples to (i.e., can heat) the pre-PCR zone  252 . 
       FIGS. 12A through 12F  illustrate an example PCR zone  254 . The PCR zone  254  thermally cycles the material between a first temperature and a second temperature, and then images the material using fluorescent markers. The imaging provides data indicating a degree of exposure to radiation of the original sample. 
     The PCR zone  254  includes a microchannel  300 . As shown in  FIG. 12A , the microchannel  300  meanders from an inlet  302  to an outlet  304 . In practice, material to be analyzed by the PCR zone  254  is pumped into the inlet  302  and flows through the microchannel  300  to the outlet  304 . 
     The PCR zone  254  includes a first heating region  310 , a detection region  320  and a second heating region  330 . One or more first heating elements  340  may extend across the first heating region  310 . Similarly, one more second heating elements  350  may extend across the second heating region  330 . The first and second heating elements  340 ,  350  may be arranged on and supported by the cover  44 . 
     The first and second heating regions  310 ,  330  are spaced away one from the other, to eliminate or reduce thermal interference between the first and second heating regions  310 ,  330 . That is, maintaining the first heating region  310  at a first temperature should not interfere with maintaining the second heating region  330  at a second temperature, and vice versa. 
     In an example, the detection region  320  is arranged between the first heating region  310  and the second heating region  330 . The detection region  320  is characterized in that there are no heating elements organized above the microchannel  300 . Other arrangements of the detection region  320 , are possible. However, placing the detection region  320  between the first and second heating regions  310 ,  330  has the advantage of efficient use of the space required for separating the first and second heating regions  310 ,  330 . 
     Additionally or alternatively to the first and second heating elements  340 ,  350 , a heating system  62  included in the bio-dosimetry device  12  can be used respectively to heat each of the first the second heating regions  310 ,  330 . 
     The microchannel  300  is formed of a plurality of interconnected segments  303  ( FIG. 12C ) each segment  303  is arranged to perform one thermal cycle on the material flowing through the microchannel  300 . One thermal cycle is defined as passing through the first heating region  310  followed by passing through the second heating region  330 . Typically, in a PCR zone  254 , the microchannel  300  may include approximately 35 segments  303 . The number of segments  303  can be adjusted, based on the specific analysis to be performed in the PCR zone  254 . 
       FIG. 12C  is an enlarged view of a section of the microchannel  300  illustrating an example segment  303   a.  The segment  303   a  has a first end  305  and a second end  307 . The segment  303   a  starts at the first end  305 , continues through the first heating region  310 , through the detection region  320 , through the second heating region  330  and back through the detector region to the second end  307 . The microchannel  300  then continues in a next segment  303   b.  As described below, when performing PCR, the first heating region  310  will be maintained at a first temperature and the second heating region  330  will be maintained at a second temperature, so that, as the material to be analyzed passes through the microchannel  300 , it is repeatedly cycled between the first temperature and the second temperature. 
     As shown in  FIG. 12C , the microchannel  300  has a width d 8 . The width d 8  can be in a range from 0.01 millimeters to 1 millimeter with a typical value of 0.05 millimeters. The width d 8  can be selected depending on a volume of fluid to be passed through the microchannel  300 , a particle size of the fluid passing through the microchannel  300 , the manufacturing process used to create the microchannel  300  in the substrate  34 , etc. 
     The microchannel  300  further has a spacing d 9  between parallel portions of microchannel  300 . The spacing d 9  can be in a range from 0.01 millimeters to 3 millimeters with a typical value of 1 millimeter and can depend on factors such as the manufacturing process used to create microchannel  300 . To conserve space in the substrate, the spacing d 9  will typically be chosen to be close to or equal to a minimum feature size allowed by the manufacturing process used to form the microchannel  300  in the substrate  34 . 
     The microchannel  300  further has a height h 3 . The height h 3  is the distance, as measured along the axis Y-Y, from a furthest extension of the microchannel  300  within the first heating region  310 , for example the point  306   a,  to a furthest extension of the microchannel  300  within the second heating region  330 , for example the point  308   a.    
     As further illustrated in  FIG. 12C , the first heating region  310  has a height h 4 . The detection region  320  has a height h 5  and the third heating region  330  has a height h 6 . The height h 3  of the microchannel  300  is equal to the sum of the height h 4  of the first heating region  310 , the height h 5  of the detector region and the height h 6  of the second heating region  330 . That is, h3=h 4 +h 5 +h 6 . 
     The height h 4  of the first heating region  310  can be determined, for example, by the area required for the first heating element  340 , based on the target first temperature of the first heating region  310 . Similarly, the height h 6  of the second heating region  330  can be determined by the area required for the second heating element  350 , based on the target second temperature for the second heating region. The height h 5  of the detection region  320  can be determined, for example, based on having a specifically defined region for detecting (counting) cDNA based on a tag emission. The sensitivity of the detection increases as an area of the detection region  320  increases. Further, as described above, in the case that the detection region  320  is located between the first heating region  310  and the second heating region  330 , the height h 5  can be determined based on a distance required to thermally isolate the first heating region  310  from the second heating region  330 . 
       FIG. 12D  is a partial side view of a section of the PCR zone  254  shown in  FIG. 12C , cut along the axis X-X. The microchannel  300  is formed in the substrate  34 . The cover  44  is attached to the substrate  34 , forming a top of the microchannel  300 . The microchannel  300  has a depth U 5 . The depth U 5  can be in a range from 10 microns to 1000 microns, with a typical value of 100 microns. 
       FIG. 12E  is a partial side view of a section of the PCR zone  254  shown in  FIG. 8C , cut along the axis Y-Y. As above, the microchannel  300  is formed in the substrate  34  and covered by the cover  44 . First and second heating elements  340 ,  350  are arranged on and supported by the cover  44 . 
       FIG. 12F  is a partial perspective view of the section of the PCR zone  254  shown in  FIG. 12D . The microchannel  300  is shown extending through the second heating region  330  and back again with each segment of the microchannel  300 . 
     In an example, the pre-PCR zone  252  may be physically arranged on the substrate  34  near (e.g., adjacent) the PCR zone  254  in the same PCR channel  250 . More specifically, the pre-PCR zone  252  may be arranged near the first heating region  310  of the PCR zone  254 . This allows the pre-PCR zone  252  to be heated to a substantially same temperature as the temperature of the first heating region of the PCR zone  254 . The heating element  260  in the pre-PCR zone  252  may be electrically coupled to the first heating element  340  in the first heating region. In a case that the heating system  62  is used to heat the first heating region  310 , the same heating system  62  can also be used to heat the pre-PCR zone  252 . 
       FIG. 13  illustrates an example organization of a cartridge  14 . A cassette  32  supports a substrate  34  in a recessed area  40 . The substrate  34  has a first side  41  and a second side  42 . A micropump  82  receives a blood sample and pumps the blood sample to the lysing region  90 . After performing lysing on the blood sample, the lysing region  90  outputs the lysed blood sample to the binding region  92 . A blister  86   b  is arranged above the binding region  92 , which may be used to inject fluid, for example, a reaction material for conversion of the mRNA to cDNA in the binding region  92 . A single blister  86   b  is only an example. One or more blisters  86   b  may be used to inject fluid into the binding region  92 . The binding region  92  outputs material including cDNA to the mixing region  94 . In the mixing region  94 , gradients of the cDNA in the material are reduced or eliminated. The mixing region  94  outputs the material including the cDNA to four pre-PCR zones  252   a,    252   b,    252   c,    252   d.  Four blisters  86   e,    86   f,    86   g,    86   h  are arranged respectively above the four pre-PCR zones  252   a,    252   b,    252   c,    252   d.  The four pre-PCR zones  252   a.    252   b,    252   c,    252   d  output material respectively to the four PCR zones  254   a,    254   b,    254   c,    254   d.  Additionally, the four blisters  86   e,    86   f,    86   g,    86   h  can output primer including fluorescent markers to the material to be analyzed respectively in the four PCR zones  254   a,    254   b,    254   c,    254   d.    
     In an embodiment, DNA based cells (bacteria and other cells) can be analyzed. In this case, a DNA trapping region can be substituted for the RNA binding region. The trapped DNA can go directly to amplification in a PCR region  96 . 
       FIG. 14  is a diagram of an exemplary process  1400  for performing bio-dosimetry on a sample with the bio-dosimetry system  10 . A sample size can be in a range of 1 to 1000 microliters and can typically be approximately 100 microliters. An example sample is a drop of blood. The process  1400  begins in a block  1402 . 
     In the block  1402 , the computer  50  is programmed to initiate bio-dosimetry on a sample. The computer  50  may receive a trigger event, such as an input from the user via the user input device  30  to begin the bio-dosimetry process  1400 , or inputs, for example from sensors  56 , that the sample is available in the reservoir  22  and a fresh cartridge  14  has been inserted into the bio-dosimetry device  12 . Based on the trigger event, the computer  50  may instruct the micropump  82  on the cartridge  14  to pump the sample from the reservoir  22  into the lysing region  90  on the cartridge  14 . Alternatively, in a case that the cartridge  14  does not include the micropump  82 , the computer  50  may instruct a pump  64  on the bio-dosimetry device  12  to pump the sample into the lysing region  90 . The process  1400  continues in a block  1404 . 
     In the block  1404 , the computer  50  is programmed to lyse the sample according to process  1500 , described below. Lysing the sample is defined herein as breaking the sample into component parts. Upon lysing the sample, the process  1400  continues in a block  1406 . 
     In the block  1406 , the computer  50  is programmed to bind target components of the sample in the binding region  92  according to a process  1600 , described below. Target components are components of the sample that the bio-dosimetry system  10  targets to remove from the lysed sample and contain in the binding region  92 . During the binding process, target components in the lysed sample are bound to the stationary phase  202  in the binding region  92 , and other components are discarded. The process  1400  continues in a block  1408 . 
     In the block  1408 , the computer  50  is programmed to convert target components from the sample bound in the binding region  92  to a material to be analyzed by PCR according to the process  1700 , described below. For example, mRNA, bound in the binding region, can be converted to cDNA. Upon converting the bound target components into material to be analyzed by PCR, the process  1400  continues in a block  1410 . 
     In the block  1410 , the computer  50  is programmed to perform PCR analysis on the material to be analyzed according to the process  1800  described below. The process  1400  continues in a block  1412 . 
     In the block  1412 , the computer  50  is programmed to store and/or output results of the PCR analysis. The computer  50  may output data on the display  28  on the bio-dosimetry device  12 . Additionally or alternatively, the computer  50  may transmit the data, for example via wireless communications to another computer. Still further, the computer  50  may be programmed to store the data from the PCR analysis for retrieval at a future time. Upon outputting and/or storing the data, the process  900  may end. 
       FIG. 15  is a diagram of an exemplary process  1500  for lysing the sample received from the bio-dosimetry device  12  in an example lysing region  90 . The sample to be lysed may be a drop of blood. Alternatively, the lysing region may be used to lyse other biological samples such as gram positive bacteria, gram negative bacteria, and encapsulated bacteria such as Anthrax. The process  1500  can be initiated by block  1404  of process  1400  and begins in the block  1502 . 
     In the block  1502 , the computer  50  is programmed to pump the sample through the lysing region  90 . For example, the sample may be a drop of blood or other liquid with a size of 100 microliters. A micropump  82  on the cartridge  14  may perform the pumping ( FIG. 4 ). Alternatively, a pump  64  in the bio-dosimetry device  12  can perform the pumping. As an example, the sample can be pumped at a flow rate corresponding to a resonance time within the cavity  160  of the lysing array  134  in a range of 5 to 30 seconds. With an average resonance time of 20 seconds, and a cavity volume of four microliters, a sample can be lysed within a time of five minutes or less. The process  1500  continues in a block  1504 . 
     In the block  1504 , which can occur concurrently with all or a portion the block  1502 , the computer  50  is programmed to apply ultrasonic stimulation to the lysing array  134 . The lysing array  134  may include a piezo electric layer  166  as shown in  FIG. 6C . Alternatively, the lysing array  134  may be coupled to a piezo electric element  68 , included in the bio-dosimetry device  12 , as shown in  FIG. 6D . The computer  50  is programmed to activate the piezo electric layer  166  or piezo electric element  68  to induce flexing of the diaphragm  155  in the lysing array  134 . At points of maximum flexing, for example near a center of the diaphragm  155 , a displacement of the diaphragm  155  may be in a range from one to 100 microns, with a typical displacement of 5 microns, and may be in a direction substantially perpendicular to a plane of the diaphragm  155 . The induced movement can typically be at a frequency of 20 kHz, with a range between 10 kHz and 40 kHz. The range is not intended to be limiting, and other frequencies above or below this range can also be used. 
     Based on the induced flexing of the diaphragm  155 , the micropillars  162  will be induced to move laterally, back and forth through their respective non-stimulated positions in the lysing array  134 . The induced lateral motion of the micropillars  162  can transfer mechanical energy to the sample, causing the sample to break into component parts. In an example, the sample includes human blood. Blood cells (leukocytes and/or erythrocytes) can be broken up to free mRNA from the cells. The process  1500  continues in a block  1506 . 
     In the block  1506 , that can occur concurrently with all or a portion of the blocks  1502  and  1504 , the computer  50  is programmed to output the lysed sample to the binding region  92 . Based on pressure from one of the micropump  82  and the pump  64 , as described with regard to block  1502 , the lysed sample passes through the outlet manifold  136 , through a microchannel formed in the substrate, and into the binding region  92 . The process  1500  ends. 
       FIG. 16  is a diagram of an exemplary process  1600  for binding components of the sample in the binding region  92 . The process  1600  can be initiated by block  1406  of process  1400  and begins in a block  1602 . 
     In the block  1602 , the computer  50  is programmed to pump the lysed sample through the binding region  92 . The block  1602  can occur concurrently with all or a portion of the process  1500 , as described above. That is, one of the micropump  82  or the pump  64  can pump the sample into the lysing region  90  and continue pumping such that the sample continues to flow into the binding region  92 . As described above, a volume of the sample in a typical case can be 100 microliters. The volume of the lysing array  134  can be in a range of four microliters. Accordingly, a portion of the sample can have exited the lysing region  90  and entered the binding region  92  while another portion of the sample has not yet entered the lysing region  90 , such that a continuous flow exists from the reservoir  22 , through the lysing region  90  and through the binding region  92 . 
     As the lysed sample flows through the binding region  92 , the binding region  92  binds to target components in the sample, containing the target components. As described above, the binding region  92  includes a stationary phase  202 . The stationary phase  202  includes a plurality of micropillars  220 . The micropillars  220  may be coated with a binding material that binds with target components in the lysed sample. 
     As the lysed sample flows through the binding region  92 , the target components bind with the binding material coated on the micropillars  220 . During the block  1602 , a sufficient quantity of the lysed sample can be passed through the binding region  92  such that the binding material on the stationary phase  202  is saturated. That is, substantially every site available to bind with a target component is occupied by a target component. As an example, the target components may be mRNA cells. The binding material may be oligo dT25 that specifically binds to any mRNA present in the sample. An amount of lysed sample through the binding region  92  to saturate the stationary phase  202  for a set of target components can be determined empirically. For example, for the binding region  92  as described above, including a stationary phase  202  coated with oligo dT25, a quantity of lysed sample of 60 microliters was determined to be sufficient to saturate the stationary phase  202 . 
     As the sample continues to be pumped through the lysing region  90  and the binding region  92 , waste material, after passing through the binding region, can be discarded. The computer  50  can be further programmed to turn off the micropump  82  or pump  64  when sufficient material has been passed through the binding region to saturate the stationary phase  202 . The process  1600  continues in a block  1604 . 
     In the block  1604 , the computer  50  is programmed to rinse the binding region  92 . The computer  50  sends instructions to force a rinsing fluid through the binding region  92 . For example, the rinsing fluid may be contained in a blister  86  arranged at the inlet  203  of the binding region  92 . The computer  50  may instruct a blister actuator  66  in the bio-dosimetry device  12  to actuate the blister  86 . The blister actuator  66  may extend a plunger into the blister  86 , expelling contents of the blister  86  into the binding region  92 . The rinsing fluid may be selected to be a material that mixes with the non-bound material in the binding region but does not react with the target components. In this manner, the target components can remain bound in the binding region  92  after rinsing. The process  1600  continues in a block  1606 . 
     In the block  1606 , which can occur during all or a portion of the block  1604 , the non-bound material, mixed with the rinsing fluid is discarded. The process  1600  ends. 
       FIG. 17  is a diagram of an exemplary process  1700  for converting the target components of the sample, bound in the binding region  92 , to material for analysis. The process  1700  can be initiated by block  1408  of process  1400  and begins in a block  1702 . 
     In the block  1702 , the computer  50  is programmed to flush the binding region  92  with a reaction mixture. For example, the cartridge  14  can include a blister  86   b  containing the reaction mixture fluidly coupled to the inlet  203  of the binding region  92 . The computer  50  can instruct a blister actuator  66  to actuate the blister  86   b  such the reaction mixture is forced into and through the binding region  92 . The reaction mixture can include the components necessary to convert the bond target components in the binding region  92  to material to be analyzed. 
     In an example, the target components may be mRNA from the sample. The reaction mixture can include the components necessary to convert, through reverse transcription, the mRNA to cDNA which can then be subjected to PCR analysis. The process  1700  continues in a block  1704 . 
     In the block  1704 , components in the reaction mixture react with the target components that are bound in the binding region  92 , forming material to be analyzed. In some cases, heat may be applied to the binding region  92  to cause/support the reaction. For example, in the case of forming cDNA from mRNA bound in the binding region  92 , the computer  50  cay be programmed to heat the binding region 92 to 50° C. for 30 minutes. The newly formed material may be bound to the stationary phase  202  of the binding region  92 . The process  1700  continues in a block  1706 . 
     In the block  1706 , the computer  50  is programmed to release the material to be analyzed for analysis. For example, in the case that the material to be analyzed is cDNA, the cDNA may be bound to the stationary phase  202 . In this case, the computer  50  can be programmed to heat the binding region to 84° C. for 5 minutes+sufficient time to release the cDNA. A time that is sufficient to release the cDNA can be determined empirically by performing tests with sample cartridges  14 . The process  1700  continues in a block  1708 . 
     In the block  1708 , the computer  50  is programmed to remove gradients of the material to be analyzed in the reaction mixture. The computer  50  sends commands to pump the reaction mixture through the mixing region  94 . As the reaction mixture flows through the mixing region  94 , the material to be analyzed, for example, cDNA, is distributed throughout the reaction mixture, reducing or eliminating gradients of the cDNA. The process  1700  ends. 
       FIG. 18  is a diagram for a process  1800  for performing qPCR analysis on the material to be analyzed. The process  1800  includes thermally cycling the material to be analyzed between a first temperature and a second temperature, and then imaging the material to be analyzed using fluorescent markers. The process  1800  can be initiated by block  1410  of process  1400  and begins in a block  1802 . 
     In the block  1802 , the computer  50  is programmed to distribute the reaction mixture including the material to be analyzed to four PCR channels  250   a,    250   b,    250   c,    250   d  for analysis. The four PCR channels  250   a,    250   b,    250   c,    250   d  include respectively four pre-PCR zones,  252   a,    252   b,    252   c,    252   d.  As shown in  FIG. 9 , the four pre-PCR zones  252   a,    252   b,    252   c,    252   d  (collectively pre-PCR zones  252 ) may be associated respectively with the four PCR zones  254   a,    254   b,    254   c,    254   d.  The computer  50  can activate a pump  64  to pump the reaction mixture into the pre-PCR zones  252   a,    252   b,    252   c,    252   d.    
     In an example, the microchannel  85  coupling the mixing region  94  to the four pre-PCR zones  252   a.    252   b,    252   c,    252   e  can split into four sub-channels  98   a,    98   b,    98   c,    98   d.  The reaction mixture including the material to be analyzed can be pumped through the microchannel, and divide between the four sub-channels  98   a,    98   b,    98   c,    98   d,  under pressure from the pump  64 , such that a portion of the reaction mixture including the material to be analyzed is directed toward each of the four pre-PCR zones  252   a,    252   b,    252   c,    252   d.    
     The process  1800  continues in a block  1804 . 
     In the block  1804 , the computer  50  may be programmed to heat the reaction mixture including the material to be analyzed in the pre-PCR zones  252   a,    252   b,    252   c,    252   d.  For example, the material to be analyzed may include cDNA. The computer  50  may be programmed to heat the reaction mixture including the material to be analyzed, to separate the cDNA into single strands prior to adding the primers and markers. The computer  50  may be programmed to heat the reaction mixture to 95° C. for 10 minutes. Other temperatures and times can be used for different materials to be analyzed. The times and temperatures can be determined empirically. The process continues in a block  1806 . 
     In the block  1806 , the computer  50  is programmed to add a set of markers and primers to the reaction mixture flowing respectively into each of the four PCR zones  254   a,    254   b,    254   c,    254   d.  A different set of markers may be used respectively for each of the four PCR zones  254   a,    254   b,    254   c,    254   d.  For example, as shown in  FIG. 9 , a first blister  86   e  may be coupled to add markers and primers to a microchannel between the pre-PCR zone  252   a  and PCR zone  254   a,  a second blister  86   f  may be coupled to add markers and primers to a microchannel between the pre-PCR zone  252   b  and PCR zone  254   b,  a third blister  86   g  may be coupled to add markers and primers to a microchannel between the pre-PCR zone  252   c  and the PCR zone  254   c  and a fourth blister  86   h  may be coupled to add markers and primers to a microchannel between the pre-PCR zone  252   d  and the PCR zone  254   d.  At a time that the reaction mixture including the material to be analyzed is flowing from the pre-PCR zone  252   a  to PCR zone  254   a,  the computer  50  may actuate the first blister  86   e.  At a time that the reaction mixture including the material to be analyzed is flowing from the pre-PCR zone  252   b  to PCR zone  254   b,  the computer  50  may actuate a second blister  86   f.  Similarly, the computer  50  may actuate a third blister  86   g  at a time that the reaction mixture including the material to be analyzed is flowing from the pre-PCR zone  252   c  to PCR zone  254   c  and a fourth blister  86   h  at a time that the reaction mixture including the material to be analyzed is flowing from the pre-PCR zone  252   d  to PCR zone  254   d.  The times that the reaction mixture is flowing respectively from pre-PCR zone  252   a  to PCR zone  254   a,  from pre-PCR zone  252   b  to PCR zone  254   b,  from pre-PCR zone  252   c  to PCR zone  254   c  and from pre-PCR zone  252   d  to PCR zone  254   d  may be the same, partially overlapping, or different, depending on an organization of the microchannel and four sub-channels between the mixing region  94  and the four pre-PCR zones  252 . For example, each of the four sub-channels coupling the respective pre-PCR zones  252   a,    252   b,    252   c,    252   d  to the microchannel coming from the mixing region  94 , may be a same length or a different length. The lengths of the sub-channels may impact a timing for adding the sets of markers to the reaction mixture for the respective PCR zones  254   a,    254   b,    254   c,    254   d.    
     The primers for each PCR zone  254  are selected to mark prepare a particular component or pair of components in the material to be analyzed for qPCR analysis. For example, for each of the four PCR zones  254   a,    254   b,    254   c,    254   d,  respective primers containing fluorescent markers may be selected to allow two genes (or DNA sequences) to be evaluated simultaneously. One gene may be an endogenous control (EC) gene that is stable with regard to exposure to radiation. An example EC gene is RB1. A second gene may be a radiation response gene. Examples of radiation response genes include CDKN1A, ASTN2, and GDF15. The process continues in a block  1808 . 
     In the block  1808 , the computer  50  is programmed to pump the reaction mixture, including the material to be analyzed and combined with the primers, into the respective PCR zones  254   a,    254   b,    254   c,    254   d.  The blocks  1808  through  1816  will be described with respect to an example PCR zone  254 . It can be understood that the process of block  1808  can be applied similarly to each of the PCR zones  254   a,    254   b,    254   c,    254   d.    
     The computer  50  may, for example, instruct a pump  64  in the bio-dosimetry device  12  to pump the reaction mixture including the material to be analyzed and the primers into the example PCR zone  254 . As described above, the PCR zone  254  includes a microchannel  300 . The microchannel  300  is formed of a plurality of segments. Each segment extends back and forth between a first heating region  310 , a detection region  320  and a second heating region  330 . In an example, the microchannel  300  includes 35 segments. 
     As an example, the material to be analyzed includes cDNA. The computer  50  may be programmed to maintain a first temperature of the first heating region at 95° C. and to maintain second temperature of the second heating region at 60° C. As the reaction mixture together with the material to be analyzed and the primer passes through the first heating region  310 , the 95° C. temperature causes the cDNA to separate into single strands. As the reaction mixture together with the material to be analyzed and the primer passes through the second heating region  330 , the 60° C. temperature allows single stranded primers to attach to single stranded DNA to form complementary strands. As the reaction mixture continues to pass through the segments of the microchannel  300 , the material to be analyzed continues to thermally cycle, that is, to cycle between the first and second heating regions  310 ,  330  at a rate of one cycle per segment, such that, with each consecutive segment, a greater number of strands of the cDNA form complementary strands with the primer. 
     The computer  50  can be programmed to continue to pump the reaction mixture in the PCR zone  254  until all or most of the microchannel  300  is filled with the reaction mixture. In this case, the farther through the microchannel  300  the reaction mixture has passed, the greater number of cycles of the first and second heating regions the reaction mixture will have experienced. The process  1800  continues in a block  1810 . 
     In the block  1810 , the computer  50  is programmed to illuminate the microchannel  300  in the PCR zone  254 . The computer  50  may radiate the detection region  320  of the PCR zone  254  with light for a time. The time may be in a range of from 1 to 2000 seconds, with a typical time of 1200 seconds. The process  1800  continues in a block  1812 . 
     In the block  1812 , the computer  50  receives images from the detection region  320  of the PCR zone  254 . The fluorescent markers that combined with the cDNA will fluoresce at wavelengths determined by the markers. The optical detector  60  in the bio-dosimetry device  12  receives the light fluoresced from the markers. Before being received by the optical detector  60 , a filter may be used to select which wavelength of light can be received by the optical detector  60 . As described above, in the case that different markers were used for two different components of the material to be analyzed (e.g., two different genes), a first image can be received from the detection region  320  with a first filter for a first range of wavelengths and a second image can be received from the detection region  320  with a second filter for a second range of wavelengths. The optical detector  60  can provide data representing the first and second images to the computer  50 . The process  1800  continues in a block  1814 . 
     In the block  1814 , the computer  50  analyzes the data of the first and second images. For example, the computer  50  may determine the Ct of each of the genes combined with a respective fluorescent marker. The Ct (cycle threshold) is defined as the number of cycles required for the fluorescent signal to exceed background level. The number of cycles required for the fluorescent signal to exceed the background level can be determined by a position within the PCR detection region  320  along an axis of increasing cycles that shows a radiation of light at the wavelength of the particular fluorescent material that exceeds the radiation of the background. The position can be directly correlated to a number of cycles to which the material to be analyzed (i.e., the genes with the fluorescent material currently being measured) were exposed before their collective illumination exceeded the background illumination. A measurement taken with an example PCR zone is described below in reference to  FIGS. 22A, 22B, 22C . The process  1800  continues in a block  1816 . 
     In the block  1816 , the computer  50  reports and/or stores the results of the PCR analysis. The computer  50  may report the results to another computer and/or on a display such as the display  28  on the bio-dosimetry device  12 . Further, the computer  50  may store the results, for example with a time stamp, or an identification of the cartridge that was measured, in memory associated with the computer  50 . The process  1800  ends. 
     As used herein, the adverb “substantially” modifying an adjective means that a shape, structure, measurement, value, calculation, etc. may deviate from an exact described geometry, distance, measurement, value, calculation, etc., because of imperfections in materials, machining, manufacturing, data collector measurements, computations, processing time, communications time, etc. 
     As an example, the bio-dosimetry system  10  can be used to determine radiation exposure of an individual. A drop of blood from the individual can be evaluated to determine the radiation dose they received based on their gene expression levels. As evidence of efficacy ex vivo irradiation of human blood was utilized and the subsequent culturing of that blood for quantitative study of utilizing gene expression for human dosimetry. 
     Selection of valid control genes is critical to have a point of reference to evaluate radiation responsive genes. Several very stable endogenous control (EC) gene products were identified. As an example,  FIG. 19A  illustrates a fold increase in expression level as a function of dose for the gene RB1 at 12 hours (trace  1902 ), 24 hours (trace  1904  and 48 hours (trace  1906 ). This EC gene is stable with respect to dose and time. It is also stable in its expression across all donors in our study. 
     Having selected a series of EC genes as controls, radiation responsive genes can be evaluated by examining the gene expression level of a potential response gene relative to that of a selected EC gene for exposed sample. A fold increase in expression levels as a function of dose and time following exposure can be determined by comparing that ratio to the population average of the non-exposed ratio of those same genes. The results for three potential radiation response genes (CDKN1A, ASTN2, and GDF15) that can be utilized as dosimetry biomarkers as shown respectively in  FIGS. 19B, 19C and 19D  referenced to RB 1 . 
       FIG. 19B  illustrates a fold increase of expression level as a function of dose for the gene CDKN1A at 12 hours (trace  1912 ), 24 hours (trace  1914 ) and 48 hours (trace  1916 ). 
       FIG. 19C  illustrates a fold increase of expression level for the gene ASTN2 at 12 hours (trace  1922 ) 24 hours (trace  1924 ) and 48 hours (trace  1926 ). 
       FIG. 19D  illustrates a fold increase of expression level as a function of dose for the gene GDF15 at 12 hours (trace  1932 ), 24 hours (trace  1934 ) and 48 hours (trace  1936 ). 
     For each of these genes CDKN1A, ASTN2 and GDF15, the curves show clear increases in expression for each of the times post irradiation examined. In a case of a nuclear event, the time of the nuclear event is known. A user can, in this case, follow the appropriate dosimetry curve for each gene product. All the gene products shown, and many others, exhibit significant increases in genes expression even at doses as low as 0.5 Gy. In the case of some genes, up to 16-fold increases of expression can be observed at higher doses. Fold changes in gene expression of 1.5 or 2 are large can easily and accurately be detected by qPCR analysis, indicating that these changes will be readily detectable by a portable PCR system. 
     In the case of several radiation responsive genes such as CDKN1A ( FIG. 19B ) gene expression levels slowly increase with time post-exposure and were highest at the longest time point examined in our studies. While other gene products such as ASTN2 and GDF15 ( FIGS. 19C and 19D ) rapidly increase at very short times post exposure and then decrease with time, they still maintain an easily-detectable increase in expression out to 48 hours post exposure. Also of note is that while a number of gene products such as CDKN1A and GDF15 ( FIGS. 19B and 14D ) show a response to radiation over the entire range of doses examined, other gene products such as ASTN2 ( FIG. 19C ) show a relatively steep dose response over a short range of doses and plateau in their expression at higher doses. This wide range in dose and time responses observed in the radiation response genes identified provides the ability to select a series of genes that can be utilized and optimized for dosimetry at various exposure levels and times post exposure. 
       FIG. 19E  illustrates a fold increase of expression of the gene CDKN1A v. dose for a in vivo radiation utilizing an animal model (C57BI/6 male mice). Traces  1941 ,  1943  and  1945  show the fold increase in expression of CDKN1A v. dose respectively at 1 day, 3 days and 7 days. Although direct comparisons of gene expression levels between humans and mice cannot be made, the measurements indicate that fold increases in CDKN1A expression due to radiation exposure in vivo can be detected in the animal models. 
       FIG. 19F  illustrates a comparison between human cancer patients irradiated in vivo and blood samples from these patients irradiated ex vivo. Sample of blood was drawn from the patients immediately prior to the patient receiving total body irradiation doses of between  2  and 4 Gy from a Cobalt 60 source A portion of the pre-radiation therapy blood sample was then irradiated with the same source and conditions as that received by the patient, and this blood was then cultured in the absence of mitogen for 24 hours. A second blood sample was drawn from the patient at approximately 24 hours post exposure. An in vivo/ex vivo exposure comparison for one of the genes examined (CDKN1A) was referenced to DPM1. Similar increase in gene expression can be observed in both the in vivo and ex vivo irradiated samples for each patient. In this study (#501, #505, #507) were given higher levels of chemotherapy while the later patients (#509, #510, #511, #513) were given lower doses of chemotherapy in their treatment. In these later patients (#509, #510, #511, #513), increases in gene expression from in vivo exposure closely match those seen in ex vivo exposure for CDKN1A shown as well as for other genes examined, and in several cases such as CDKN1A expression increases match those seen in samples from healthy donors. The fact that gene expression levels for several genes, which could be utilized for radiation dosimetry, remain viable as biomarkers even in cancer patients receiving light doses of chemotherapy is a good indication of the robustness of these markers and to their potential resistance to confounding factors in the general population. 
     Both the results from our mouse study and our evaluation of TBI patients support the validity of the use of ex vivo irradiated blood as a valid model for radiation exposure in humans for the study of gene expression from a blood sample. The animal study showed that comparable gene expression increases, resulting from radiation exposure, occur in an in vivo irradiation animal model as those seen with the ex vivo irradiation of cultured blood samples. The animal study further confirms that gene expression is a valid and feasible dosimetry method even at times of seven days post exposure and possible beyond. In vivo/ex vivo comparisons of gene expression levels in human cancer patients confirm that increases in gene expression due to radiation exposure for the genes examined in our study are not dependent on whether the radiation exposure was performed in vivo or ex vivo. 
     The results from individual genes as shown in  FIGS. 19A-19F  illustrate a time dose dependence to the genes expression levels. To accurately perform dosimetry based on data from these genes, a statistical model is required. To develop a statistical model, multiple predictive models to determine dosimetry were developed for the various time points post exposure. These regression models, a quadratic fit to the log of dose, were fit using forward stepwise regression. At each step a gene was added to the model based on the following: (a) the greatest contribution to increasing R-squared and (b) maintaining the available sample size. The statistical model for time point post exposure is shown in  FIGS. 20A-20C  where the actual radiation dose is plotted against the model&#39;s predicted dose. The genes utilized in this model were CDKN1A, ATM, ASTN2, and GDF15.  FIG. 20A-20C  compare modeling results with ex vivo human data respectively for 12 hours, 24 hours and 48 hours post exposure. 
     While the modeling of the gene expression data is modeled with respect to the log of dose, due to the exponential nature of Ct values, this data is converted to a linear scale for the plots in  FIGS. 20A-20C . It is this conversion from the native log scale of the model to a linear dose for plotting that makes the data at high doses in  FIGS. 20A-20C  appears to be more scattered then at the lower doses. 
     An example result is shown in  FIG. 20D  where a receiver operating characteristic (ROC) curve for a prediction of exposure of 2 Gy or greater is shown at 12 hours post exposure. For the data shown in  FIG. 20D , the area under the ROC curve was 0.9816 (value of 1 being perfect), which is very good for biomedical screening application. Similar ROC curves were obtained for the predictive models for the other post exposure time points examined in the ex vivo human study and for the data from the animal model. In order to further reduce any false negatives, sensitivity could be maximized at the cost specificity resulting in increased false positives. 
     Tests were performed on an example lysing device to estimate the lysing capability of the lysing region  90  described above. The example lysing device used for the test had included 1600 micropillars arranged in a 40 by 40 lysing array. The micropillars were supported on a 6 mm×6 mm×0.1 mm thick diaphragm. The micropillars had a diameter of 0.075 mm and a height of 0.75 mm. The spacing between the micropillars was 0.075 mm. To estimate the effectiveness of the lysing region, PCR analysis was performed on four different samples exposed to varying degrees of lysing. The results are of the tests are shown in  FIG. 21 , which illustrates example traces of cycle threshold (Ct) from PCR analysis of the four different blood samples. The Ct level indicates a number of remaining unlysed lymphocites in each blood sample. 
     Trace  2102  in  FIG. 21  is an example trace from PCR analysis of whole blood that is passed through an example lysing device without powering the lysing device. 
     Trace  2104  is an example trace from PCR analysis of whole blood that was not passed through the lysing device (a sample representing the condition of the blood as input into the lysing device). 
     Trace  2106  is an example trace from PCR analysis of a control_lynmphocyte GM40. The control lymphocyte is a set amount that is injected as a known control for reference. 
     Trace  2108  is an example trace from PCR analysis of whole blood that is passed through the lysing device under power. That is, the diaphragm is actuated at 20 kHz while the whole blood is passed through the lysing device. 
       FIGS. 22A, 22B, 22C  illustrate an example image of a PCR zone  254  containing material to be analyzed that has been illuminated and is fluorescing. As described above, the PCR zone  254  includes a microchannel  300  including a plurality of segments. Each segment cycles through a first heating region  310 , a detection region  320  and a second heating region  330 . 
     In the test, material to be analyzed is pumped into the microchannel  300  in the PCR zone  254 . In an example, sufficient material can be pumped into the microchannel  300  that all of the plurality of segments contain material. When, the microchannel  300  is filled in this manner, the detection region  320  in the PCR zone  254  can be illuminated. Following illumination, an image can be taken of the detection region  320 . The resulting image can show, by an intensity of fluorescent light at locations (i.e., in different segments) of the microchannel  300 , the cycle threshold (Ct) required to detect a component (for example, a gene) within the material to be analyzed. The cycle threshold (Ct) is defined as the number of cycles (through the first and second heating regions  310 ,  330 ) required for the fluorescent signal from the tag to exceed background level. The cycle threshold provides an indication of a degree of exposure to radiation of the original sample used to prepare the material to be analyzed. 
     In  FIGS. 22A, 22B, 22C , the number of cycles of the PCR to which the material to be analyzed has been exposed increases from left to right.  FIG. 22A  is a beginning (left) portion of the PCR zone  254 .  FIG. 22B  is a middle portion of the PCR zone  254 .  FIG. 22C  is an end (right) portion of the PCR zone  254 . Imaging is performed in the detection region  320  between the first and second heating regions  310 ,  330  (see, for example,  FIGS. 12A and 12C ). In the area labelled  2202 , the fluorescing signal from the material to be analyzed begins to exceed the background level. This is indicated by the dark oval shaped areas  2203  in the middle of the image. As can be seen, the intensity of the fluorescing signal continues to increase from left to right. This increasing discontinues at approximately a point  2204 . This indicates that the material to be analyzed was only pumped into the PCR zone up to this point. 
       FIG. 23  shows an example amplification of two targeted genes with a PCR. The first target gene was cDNA made from human 18S rRNA. The sample was run through the sample PCR and yielded a Ct (PCR cycled threshold) value of 13.7 (trace  2302 ). The same sample was then diluted by a factor of 1 to 1000 and run on the PCR. This dilution should correspond to a Ct shift of ˜10 cycles. The resulting trace  2304  corresponds well with this expected shift. 
     With regard to the media, processes, systems, methods, etc. described herein, it should be understood that, although the steps of such processes, etc. have been described as occurring according to a certain ordered sequence, such processes could be practiced with the described steps performed in an order other than the order described herein. It further should be understood that certain steps could be performed simultaneously, that other steps could be added, or that certain steps described herein could be omitted. In other words, the descriptions of systems and/or processes herein are provided for the purpose of illustrating certain embodiments and should in no way be construed so as to limit the disclosed subject matter. 
     Accordingly, it is to be understood that the present disclosure, including the above description and the accompanying figures and below claims, is intended to be illustrative and not restrictive. Many embodiments and applications other than the examples provided would be apparent to those of skill in the art upon reading the above description. The scope of the invention should be determined, not with reference to the above description, but should instead be determined with reference to claims appended hereto and/or included in a non-provisional patent application based hereon, along with the full scope of equivalents to which such claims are entitled. It is anticipated and intended that future developments will occur in the arts discussed herein, and that the disclosed systems and methods will be incorporated into such future embodiments. In sum, it should be understood that the disclosed subject matter is capable of modification and variation. 
     The article “a” modifying a noun should be understood as meaning one or more unless stated otherwise, or context requires otherwise. The phrase “based on” encompasses being partly or entirely based on.