Patent Publication Number: US-2019168213-A1

Title: System and method for determining efficacy and dosage using parallel/serial dual microfluidic chip

Description:
CROSS-REFERENCE TO RELATED APPLICATIONS 
     This application claims priority to and the benefit of U.S. Provisional Application No. 62/584,653, filed Nov. 10, 2017, and entitled SYSTEM AND METHOD FOR DETERMINING EFFICACY AND DOSAGE USING PARALLEL/SERIAL DUAL MICROFLUIDIC CHIP, the contents of which are incorporated by reference herein in their entirety. 
    
    
     TECHNICAL FIELD 
     The present invention pertains in general to a microfluidics lab-on-chip system and, more particularly, to the use of a microfluidics chip and testing at the point of care. 
     BACKGROUND 
     The emergence and spread of antibiotic-resistant bacteria are aggravated by incorrect prescription and use of antibiotics. Courts have this problem is the fact that there is no sufficiently fast diagnostic test to guide correct antibiotic prescription at the point of care. Currently, some fluid sample is retrieved from a patient and forwarded to a lab for testing to determine a specific treatment regimen. As a safeguard, the patient is sometimes initially given large doses of a general antibiotic until a more specific antibiotic can be determined to target the specific bacteria. This can take upwards of two or three days, as the process requires growing the bacteria in some culture medium and observing its response to various antibiotics. 
     SUMMARY 
     The present invention disclosed and claimed herein, in one aspect, comprises a method for determining a treatment agent and dosage level for a predetermined biologic material receives a biologic sample containing the predetermined biologic material for treatment by one of a plurality of treatment agents. The biologic sample containing the predetermined biologic material is held within a first reservoir. A portion of the biologic sample is pumped into each of a first plurality of parallel pathways from the first reservoir using a micro-pump. A separate treatment agent of the plurality of treatment agents is applied within each of the first plurality of parallel pathways to the portion of the biologic sample within the parallel pathway. The treatment agent of the plurality of treatment agents providing a best treatment efficacy for the predetermined biologic material within the biologic sample is determined responsive to the plurality of treatment agents applied to the portion of the biologic sample within each of the first plurality of parallel pathways. A second portion of the biologic sample is pumped into a selected second parallel pathway associated with the determined treatment agent of a second plurality of parallel pathways from the first reservoir using a second micro-pump. The determined treatment agent at a plurality of different dosage levels is applied within the selected second parallel pathway to the second portion of the biologic sample within the second parallel pathway. A dosage level of the plurality of different dosage levels of the determined treatment agent is determined with respect to the predetermined biologic material providing the best treatment efficacy. An output indicating the treatment agent and the dosage level of the treatment agent providing the best treatment efficacy is then provided. 
    
    
     
       BRIEF DESCRIPTION OF THE DRAWINGS 
       For a more complete understanding, reference is now made to the following description taken in conjunction with the accompanying Drawings in which: 
         FIG. 1  illustrates a high-level view of a microfluidics chip of the present disclosure; 
         FIGS. 2A-2C  illustrate detailed views of the multiple stages of analysis provided by the microfluidics chip of  FIG. 1 ; 
         FIGS. 3A-3D  illustrate diagrammatic views of the various cell capture regions and the interspersed pumps for the microfluidics chip of  FIG. 1 ; 
         FIGS. 4A-4G  illustrates detailed views of the first viewing stage; 
         FIG. 5  illustrates a detailed view of the first parallel driving stage; 
         FIGS. 5A and 5B  illustrate details of the coating applied to the micro channels in the first driving stage; 
         FIG. 6  illustrates a detail of the serial driving stage; 
         FIGS. 7A-7D  illustrate detailed views of a valveless nozzle/diffuser micropump; 
         FIG. 8  illustrates a detailed view of a piezoelectric micropump; 
         FIG. 9  illustrates a detailed view of a multi-chamber micropump with check valves; 
         FIG. 10  illustrates a flowchart for the high-level operation of the microfluidics chip; 
         FIG. 11  illustrates a flowchart for the initial loading operation of the fluid sample; 
         FIG. 12  illustrates a flowchart for the viewing or cell counter stage of analysis; 
         FIGS. 13A-13C  illustrate diagrammatic use for the cell counter; 
         FIG. 14  illustrates a flowchart for the main parallel stage of analysis; 
         FIG. 15  illustrates the serial stage of analysis; 
         FIG. 16  illustrates a simple fight diagrammatic view of the microfluidics chip; 
         FIG. 17  illustrates a simplified diagrammatic view of a parallel module; 
         FIG. 18  illustrates simplified diagrammatic view of a serial module; 
         FIG. 19  illustrates a simplified diagrammatic view of a serial module arranged in parallel; 
         FIGS. 20A and 20B  illustrated a diagrammatic view of an embodiment utilizing a chemostat; 
         FIG. 21  illustrates a diagrammatic you have the microfluidics chip utilizing valves; 
         FIGS. 22A-22B  illustrate cross-sectional views of a micro valve 
         FIG. 23  illustrates a diagrammatic view of preparing a biologic sample and disposing it in the well on the microfluidic chip; 
         FIG. 24  illustrates a cross-sectional view of an RT-lamp interfaced with a cell phone; 
         FIG. 25  illustrates a perspective view of the RT lamp interfaced with a microfluidic chip and a cell phone; 
         FIG. 26  illustrates a side view of a cell phone interfacing with the micro fluidic chip; 
         FIG. 27  illustrates a window view of the camera and the alignment process; 
         FIGS. 28A-28H  illustrate multiple views of a diagram of the microfluidic chip in schematic form and various loading and analysis steps associated there with; 
         FIG. 29  illustrates a flowchart for the overall analysis process utilizing the microfluidic chip; and 
         FIG. 30  illustrates a flowchart to pick in the details of the test path. 
     
    
    
     DETAILED DESCRIPTION 
     Referring now to the drawings, wherein like reference numbers are used herein to designate like elements throughout, the various views and embodiments of a System and Method for Determining Efficacy and Dosage Using Parallel/Serial Dual Microfluidic Chip is illustrated and described, and other possible embodiments are described. The figures are not necessarily drawn to scale, and in some instances the drawings have been exaggerated and/or simplified in places for illustrative purposes only. One of ordinary skill in the art will appreciate the many possible applications and variations based on the following examples of possible embodiments. 
     Referring now to  FIG. 1 , there is illustrated a diagrammatic view of a microfluidics chip  102  at a high-level view. There is provided in the microfluidics chip  102  an input stage  104  that is operable to receive a biological specimen. As used herein, a “sample” must be capable of flowing through microfluidic channels of the system embodiments described hereinbelow. Thus, any sample consisting of a fluid suspension, or any sample that be put into the form of a fluid suspension, that can be driven through microfluidic channels can be used in the systems and methods described herein. For example, a sample can be obtained from an animal, water source, food, soil, air, etc. If a solid sample is obtained, such as a tissue sample or soil sample, the solid sample can be liquefied or solubilized prior to subsequent introduction into the system. If a gas sample is obtained, it may be liquefied or solubilized as well. The sample may also include a liquid as the particle. For example, the sample may consist of bubbles of oil or other kinds of liquids as the particles suspended in an aqueous solution. 
     Any number of samples can be introduced into the system for analysis and testing, and should not be limited to those samples described herein. A sample can generally include any suspensions, liquids, and/or fluids having at least one type of particle, cellular, droplet, or otherwise, disposed therein. In some embodiments, a sample can be derived from an animal such as a mammal. In a preferred embodiment, the mammal can be a human. Exemplary fluid samples derived from an animal can include, but are not limited to, whole blood, sweat, tears, ear flow, sputum, bone marrow suspension, lymph, urine, brain fluid, cerebrospinal fluid, saliva, mucous, vaginal fluid, ascites, milk, secretions of the respiratory, intestinal and genitourinary tracts, and amniotic fluid. In other embodiments, exemplary samples can include fluids that are introduced into a human body and then removed again for analysis, including all forms of lavage such as antiseptic, bronchoalveolar, gastric, peritoneal, cervical, athroscopic, ductal, nasal, and ear lavages. Exemplary particles can include any particles contained within the fluids noted herein and can be both rigid and deformable. In particular, particles can include, but are not limited to, cells, alive or fixed, such as adult red blood cells, fetal red blood cells, trophoblasts, fetal fibroblasts, white blood cells, epithelial cells, tumor cells, cancer cells, hematopoeitic stem cells, bacterial cells, mammalian cells, protists, plant cells, neutrophils, T lymphocytes, CD4+, B lymphocytes, monocytes, eosinophils, natural killers, basophils, dendritic cells, circulating endothelial, antigen specific T-cells, and fungal cells; beads; viruses; organelles; droplets; liposomes; nanoparticles; and/or molecular complexes. In some embodiments, one or more particles such as cells, may stick, group, or clump together within a sample. 
     In some embodiments, a fluid sample obtained from an animal is directly applied to the system described herein at the input stage, while in other embodiments, the sample is pretreated or processed prior to being delivered to a system. For example, a fluid drawn from an animal can be treated with one or more reagents prior to delivery to the system or it can be collected into a container that is preloaded with such a reagent. Exemplary reagents can include, but are not limited to, a stabilizing reagent, a preservative, a fixant, a lysing reagent, a diluent, an anti-apoptotic reagent, an anti-coagulation reagent, an anti-thrombotic reagent, magnetic or electric property regulating reagents, a size altering reagent, a buffering reagent, an osmolality regulating reagent, a pH regulating reagent, and/or a cross-linking agent. 
     At this point in the process, a finite amount of biofluids is disposed in the reservoir ready for transferring to subsequent stages. This amount of fluid is then transferred to another stage via a driving stage  106  in order to transfer this biofluid to another reservoir, that associated with a viewing stage  108 . At this stage, a technician can examine the biofluid and determine the makeup of the biofluid, discriminate cells, etc. in order to make certain decisions as to going forward with remaining tests. The microfluidic chip then transfers the biofluid at the viewing stage  108  to a parallel analysis stage  115  through a parallel driving stage  110  wherein the biofluid is divided among a plurality of parallel path this for analysis of the reaction of the material in the biofluid with different reagents in a reading. This requires a certain amount of the biofluid to be transferred to this analysis stage. Thereafter, a decision is made as to whether to transfer the remaining biofluid from the viewing stage  108 , in order to perform more testing and/or analysis on the biofluid. At this stage the process, only one of the multiple second stage or serial stage path is selected. One reason for this is that there is only a finite amount of biofluid available and there is no need for testing along paths that are associated with previous decisions indicating that the results will be negative along these paths. Each of these serial passes associated with one of the parallel paths. Thus, if there are five parallel paths, there will be five serial paths. Note that the term “serial path” is a term meaning that it is within the serial decision tree and it need not actually be a plurality of serial paths that are linked together in a serial manner, although they could be and are in some embodiments described hereinbelow. It is necessary to perform the testing/analysis along each of the five parallel paths, but a decision at this point indicates that only one of the serial paths will be required for the testing/analysis purpose. This will be described in more detail hereinbelow. 
     Referring now to  FIGS. 2A-2C , there are illustrated diagrammatic views of the various stages of the process. With specific reference to  FIG. 2A , there is illustrated a diagrammatic view of first viewing stage, wherein the amount of biofluid stored in the input stage reservoir  104  is driven to the viewing stage  108  reservoir. At this stage, optical device  202 , for example, can be used to view the cells disposed within the medium. This medium could actually be the actual biofluid that was provided in the sample from the human/animal or could be some diluted version thereof. However, this biofluid will contain some cellular material or some particulate of interest. This can be viewed with the out device  202  and then passed to a processor  204 , or a human could analyze the results. With utilization of the processor  204 , the actual form of biofluid, and analog form, is transferred to a digital form. This could be in the form of cell counting for verification of a particular cell. As will be described hereinbelow, affinity labels can be associated with each of the cells or particulates in the biofluid and this could facilitate visual recognition of different characteristics or different types of cells, such as proteins, bacteria, etc. Each of these cellular materials can have a particular affinity label associated there with that allows it to be visually identified via some characteristics such as florescence or even magnetic properties associated with the affinity label. Again, this will be described hereinbelow. Although an optical device  202  was illustrated and described, any other type of device for analyzing the characteristics of a particular affinity labeled cell can be utilized, such as some type of magnetometer, etc. 
     Referring now to  FIG. 2B , there is illustrated the next parallel drive stage. At this stage, a micropump is utilized in the parallel drive stage  110  to pump at least a portion of the biofluid stored in the reservoir associated with the viewing stage  108  is transferred to all of the parallel reading/analysis paths. In this step, it can be seen that a portion of the biofluid in the reservoir associated with the viewing stage  108  and is biofluid exists in each of these parallel paths for analysis. There is an indication in one of these parallel paths, associated with the reservoir  210 , that shows a positive indication of a reaction of some type that is viewable. If, for example, this were bacteria, one reagent could be an antibiotic in a large dosage that would destroy the particular target bacteria and this would be recognized by an observer. The other three paths, associated with reservoirs  214 ,  216  and  218  (an example of  4  paths), would have no reaction and, as such, would not have affected the bacteria associated therewith. In this example, a high level of concentrated antibiotic is provided that would destroy the bacteria, but at this level of analysis, there is no indication provided as to the actual dosage of that antibiotic that would destroy the bacteria, other than the fact that a large dosage of this particular antibiotic will destroy the target bacteria. It is important to keep in mind that this particular biofluid may have multiple and different bacteria, proteins, etc. contained therein. 
     Referring now to  FIG. 2C , there is illustrated a diagrammatic view of the final serial stage of analysis/testing. Since the first stage of testing/analysis transferred some of the biofluid from the viewing stage  108  to the parallel stages  114 , there is still some biofluid remaining in the viewing stage  108 . This is a selectively transferred to one of the serial paths, that associated with the testing reservoir  210 . There are provided a plurality of bypass channels  220  associated with each of the serial paths and only the bypass channel  220  associated with the reservoir  210  in the parallel path  114  will be selected for transferring biofluid to this particular serial path associated with the reservoir  210  for testing. It will first be pumped to be a micropump in a serial drive stage  222  to a first serial reservoir  224  for testing/analysis. If the test is negative, it can then be passed to a subsequent serial driving stage  226  to a subsequent serial reservoir  228  for testing/analysis and so on. As will be described hereinbelow, there can be provided a single bypass path  220  which is connected to a manifold associated with each of the serial paths and each of the manifolds can be associated with each of the different reservoirs for testing, i.e., at this point the testing is parallel to all of the subsequent testing reservoirs. In the mode illustrated in this  FIG. 2C , it is necessary to transfer all of the necessary biofluid, i.e., typically the remaining biofluid in the viewing stage reservoir  108 , to the reservoir  224  and pass all of that biofluid to the next reservoir  228  and so on. Thus, at each stage, all of the biofluid transferred in the subsequent stages is tested at each subsequent stage. In a parallel configuration, the remaining biofluid in the viewing stage  108  would be required to be divided among the different testing reservoirs at each of the subsequent stages. This will be described in more detail hereinbelow. 
     Referring now to  FIGS. 3A-3D , there are illustrated diagrammatic views of the process and fluid flow. In  FIG. 3A  come there is illustrated an overall process flow for the embodiment described hereinabove. This embodiment, there is provided an input well  302  for receiving the biologic sample indicated by numeral  303 . This constitutes a finite volume that must be transferred via a micropump to a viewing reservoir  306 . At this point, substantially all of the biofluid is transferred from the reservoir  302  to the viewing reservoir  306 . This is the first stage of the process. The second stage of the process is illustrated as providing three separate testing reservoirs  308 ,  310 ,  312 , attached at one to a microchannel manifold  314 . Each of the testing reservoirs  308 ,  310 ,  312 , as will be described hereinbelow, is comprised of a serpentine microchannel  316  attached at one end to the manifold  314  and at the other end to a viewing reservoir  318 . A micropump  320  is provided for transferring biofluid from the viewing reservoir  306  to the manifold  314 . This will be divided among the three testing reservoirs  308 ,  310 ,  312  and substantially even amounts. The biofluid will traverse the serpentine microchannel  316 , which is coated with a particular reagent, one example being an antibiotic. In this example, the antibiotic is at a very high concentrated level, each of the testing reservoirs  308 ,  310  and  312  having a different antibiotic associated there with. Only a portion of the biofluid in the viewing reservoir  306  will be transferred to these three testing reservoirs  308 ,  310  and  312  for testing/analysis and viewing at the associated viewing reservoir  318 . The serpentine shape, when used with a medium containing cells such as in a biologic sample, facilitates and enhances mixing due to the increased interfacial contact area between the cells within the biofluid sample. 
     The next step of testing/analysis will be selected only upon a positive test occurring within one of the three testing reservoirs  308 ,  310  and  312 . However, each of the testing reservoirs  308 ,  310  and  312  has associated there with a subsequent group of testing reservoirs. In this embodiment, each of the subsequent testing reservoirs is comprised of a plurality of sub reservoirs  330 , each of the sub reservoirs  330  being configured identical to the testing reservoirs  308 ,  310  and  312 , with a serpentine microchannel region  316  and a viewing reservoir  318 . A single bypass microchannel  220  is provided to connect viewing reservoir  306  to a sub reservoir manifold  332 . Each of the particular sub reservoir paths have associated there with a separate micropump  334 . Only one of these micropumps  334  is selected for transferring the remaining portion of the biofluid stored in the viewing reservoir  306  to the selected path. In this embodiment, the remaining portion of the biofluid is transferred to the first reservoir  330  bypassing the biofluid through the serpentine microchannel  316  to the associated viewing reservoir  318 . This particular microchannel will have coating of antibiotic, in this example above, at a relatively low dose. If the bacteria, for example, do not react accordingly with this level of antibiotic, it can be recognized as such in the viewing reservoir  318 . It is noted that the antibiotic associated with the coating on the walls of the microchannel  330  at this dosage will not be picked up by the bacteria and, as such, the bacteria in the viewing reservoir  318  for the first sub reservoir  330  in the selected path will still be intact. It can then be pumped from the reservoir  318  associated with the first testing reservoir  330  in the chain to a subsequent testing reservoir  330  with a subsequent micropump  336 . This subsequent sub reservoir will have a concentration of antibiotic in its serpentine microchannel  316  that is at a higher level. As the level increases, a gradient is tested for, such that the dosage can be gradually increased until the bacteria are destroyed. If, for example, the bacteria were associated with an affinity label that made it fluoresce, this would be recognized. It could also be that there are multiple bacterial types contained within the biofluid that are each associated with a different affinity label and this could be recognized. It could, in fact, the case that one type of bacteria perfected at a first dosage level of the antibiotic and a second bacteria were affected at a another dosage level of the antibiotic. 
     Referring now to  FIG. 3B , there is illustrated a diagrammatic view of an alternate process flow. This will work substantially identical to the embodiment of  FIG. 3A , come up until the operation at the manifold  332  associated with the sub reservoirs. In this embodiment, the three micropumps  334  each feed a sub reservoir manifold  340 . Each of the sub reservoir manifolds  340  is connected to a plurality of the sub reservoirs  330  associated with each path. In this embodiment, there are only illustrated three sub reservoirs  330  for each of the sub reservoir manifolds  340 , although each path could have a different number of sub reservoirs  330  associated therewith. The difference between these two embodiments is that, at this point, the amount of biofluid remaining in the viewing reservoir  306  now must be divided amongst all of the sub reservoirs attached on one end thereof to the associated sub reservoir manifold  340  selected by the activated one of the micropumps  334 . This will result in potentially less biofluid being available for the testing/analysis step. This will also mean that each of the viewing reservoirs  318  associated there with will have a smaller volume associated therewith. 
     Referring now to  FIG. 3C , there is illustrated a diagrammatic view that provides a simplified diagram of the transfer from reservoir to reservoir. In this illustration, the input stage is illustrated as an input reservoir  350  labeled R 0 . A micropump  352  is operable to transfer the contents of this input reservoir, the biofluid, to a second reservoir, a viewing reservoir  354 , labeled R 1 . A portion of the contents of this reservoir are then transferred via a micropump  356  to a plurality of parallel stage reservoirs  358  labeled R 2 . This is the first testing/analysis stage. After this stage, the remaining contents of the viewing reservoir  354  are transferred to the subsequent serial stage reservoirs via a pump  360  via a bypass path and microchannel  362 . The serial stage reservoirs are labeled R 3 , R 4 , etc. This illustration sets forth how the entire contents of the input reservoir R 0  are transferred down the chain. This is best illustrated in  FIG. 3D . In this illustration, it can be seen that entire contents of reservoir R 0  are transferred to reservoir R 1 . At this point, only a portion of the contents are transferred to reservoir R 2 . The remaining contents are sequentially transferred to R 3 , R 4 , and so on. For this illustration, the entire remaining contents of the reservoir  354 , R 1 , will be transferred down the chain entirely to reservoir R 3 , then to reservoir R 4 , and so on. In the alternate embodiment, as described hereinabove, and not illustrated in  FIG. 3D , the bypass  362  could be connected to each of the reservoirs R 3 , R 4 , etc. in parallel, noting that the remaining contents of the reservoir R 1  will then be divided amongst the parallel connected reservoirs R 3 , R 4 , etc. 
     Referring now to  FIGS. 4A-4G , there are illustrated diagrammatic views of the initial processing section associated with the viewing stage  108 . There is provided a substrate  402  upon the surface of which are formed a plurality of wells and microchannels. A first well  404  is provided for receiving the biofluid sample in this well has a finite volume associated there with. At the bottom of this well a microchannel  406  extends outward and up to the surface to an opening  408 . The purpose of this microchannel  406  extending to the bottom of the well  404  is to ensure that the biofluid can be completely pumped from the well  404 . For the formation of this microchannel  406 , it might be that the microchannel is formed through the surface of the substrate  402  and then a cover plate (not shown) having a surface that extends down into the open microchannel. An adjacent channel  410  is disposed proximate the opening  408  to provide another opening therefore in order to accommodate a micropump  412  (shown in phantom) interface with the opening  408  and the one end of the microchannel  410  for transferring fluid from the well  404  to the microchannel  410 . The microchannel  410  extends along the surface of substrate  402  in order to interface with a viewing well/reservoir  412 . As the biofluid passes through the microchannel  410  and the viewing well  412 , a desired analysis can be performed on the contents of the biofluid. As described hereinabove, in one example, various cells in the biofluid might consist of different types of bacteria, proteins, etc. and each of these may have associated there with a specific affinity label, which is optically detectable. There are, of course, other means by which affinity labels can be detected. As the cells contained within the biofluid pass through the viewing well/reservoir  414 , they can be examined. The viewing well/reservoir  414  on the other side thereof is connected to one side of a microchannel  416 , the other side thereof connected to a reservoir  418 . Since the micropump  412  must force the biofluid through the microchannels and the viewing well/reservoir  414 , there is required the necessity for a holding reservoir  418  to be present. However, initially, this reservoir  418 , the microchannel  410  and the viewing well/reservoir  414  will have air disposed therein. This air must be removed. This can be done with a negative pressure of some sort or just a waste gate output to the atmosphere. This is provided by a waste gate microchannel  420  that is connected to an opening  422  through the cover glass (not shown) or to the side of substrate  402 . A valve  423  could be provided above the opening  422 . As biofluid enters the reservoir  418 , air will be pushed out through the microchannel  420 . It is desirable for this microchannel  422  to have as low a profile as necessary such that only air exits therefrom. Depending upon the size of the cells contained within the biofluid, the microchannel  420  can be significantly smaller and have a lower profile than the microchannels  410  and  416 . Is important to note that, once the micropump  412  transfers the biofluid from the well  404 , the volume transferred will be spread between the two microchannels  410  and  416 , the viewing well  414  and the reservoir  418 . Thus, the reservoir  418  has a significantly larger volume that any of the microchannels  410  and  416  and the viewing well/reservoir  414 . Additionally, it may be that the depth of the wells/reservoirs  404  and  418 , as well as the viewing well reservoir  414  are also as shallow as the microchannels  410  and  416  but significantly wider to accommodate the required volume. 
     The outlet of the reservoir  418  is connected from the bottom thereof through a microchannel  426  to an opening  428  on the upper surface of the substrate  402 . This is interfaced with a micropump  430  (in phantom) to an adjacent microchannel  432  for subsequent processing. These micropumps  412  and  430 , although illustrated as being flush with the substrate, will typically be disposed above the cover plate (not shown) with holes disposed through the cover plate. The opening  428  will be a horizontal microchannel associated with the manifold  314  described hereinabove. This will be associated with a plurality of micropumps  430  for each of the parallel paths or the bypass path. A cross-sectional view of the embodiment of  FIG. 4A  is illustrated in  FIG. 4B , with a cover plate  440  disposed over the substrate  402  with an opening  442  disposed above the well  404  for receiving the biofluid sample. 
       FIGS. 4C and 4D  illustrate top view and cross-sectional views of the reservoir  418  illustrating how the microchannel  416  feeds biofluid into the top of the reservoir  418 , and the flow path for the biofluid from the reservoir  418  through the microchannel  426  from the bottom of the reservoir  418 . However, it may be that, with capillary action, the depth of the reservoir  418  could be equal to that of the microchannels  416  and  426  such that they are all at the surface of the substrate  402  for ease of manufacturing. When a negative pressure is placed upon the reservoir  418 , air will be pulled into the microchannel  426  through the microchannel  420 . It is possible in this mode that the micropump  412  could be operated to actually create a positive pressure in the microchannel  416  to force the biofluid in the reservoir  418  into the opening  428  through the microchannel  426 . Again, the microchannel  420  would preferably have a dimension that was smaller than the smallest cell size within the biofluid. 
     Referring now to  FIGS. 4E and 4F , there are illustrated top view and cross-sectional views of the reservoir  418  with an alternate embodiment illustrating microchannel  426 ′ as being beneath the bottom of the reservoir  418  to allow more complete emptying of the reservoir  418 . 
     Referring now to  FIG. 4G , there is illustrated an alternate embodiment of inlet wells for receiving the biofluid sample. There is provided the well  404  for receiving the biofluid sample and a second well  464  receiving an additional fluid sample. This fluid sample in well  460  could be some type of dilutant or it could be a medium containing various affinity labels. As noted hereinabove, the fluid sample could have associated there with affinity labels prior to the biofluid sample being disposed in the well  404 . However, it is possible that the microfluidic chip have disposed in the well  460  a medium containing affinity labels, for example. The well  460  would be interfaced through a microchannel  462  to an opening  464  adjacent the opening  408 . A two input, one output, micropump  412 ′ that interfaces with the microchannel  410 . 
     Referring now to  FIG. 5 , there is illustrated a diagrammatic view of the microchannel structure associated with the parallel stage of operation. The microchannel  426  is interfaced with a microchannel manifold  502  which corresponds to the opening  428 . This microchannel manifold  502  is interfaced with a plurality of micropumps  504 , corresponding to the micropump  430 . These micropumps  504  are disposed in pairs, each pair associated with one testing reagent. As noted hereinabove, there are provided a plurality of parallel paths, each associated with a reservoir  312  having a serpentine microchannel  316  and a viewing reservoir  318 . The first micropump  504  in the pair of micropumps  504  is connected to one end of the associated serpentine microchannel  316 . When this micropump  504  is activated, biofluid from the reservoir  418  is passed through the manifold microchannel  502  and through the serpentine microchannel  316  to the viewing reservoir  318 . As was the case above, there is provided a waste microchannel  506  for each of the reservoirs  318  to allow air to escape therefrom as biofluid is forced through the microchannel  316 . The micropump  504  associated with this serpentine microchannel  316  will be operated for a sufficient amount of time to transfer sufficient biofluid from the reservoir  418  through the serpentine a channel  316  and finally into the reservoir  318  to fill the reservoir  318 . The microchannel  506  can have some type of valve associated with the opening thereof to prevent the escape of any biofluid therefrom or, alternatively, the dimensions of that microchannel  506  could be small enough to prevent any appreciable amount of cells escaping therefrom. Although not illustrated, the one of the pair of micropumps  504  associated with the parallel stage of operation and associated reservoirs  312  will also be operated to fill the associated serpentine microchannel  316  and reservoir  318 . 
     Referring now to  FIGS. 5A and 5B , there are illustrated cross-sectional views of the serpentine microchannel  316 . As described hereinabove, the sides of these channels  316  are coated with some type of reagent. For example, if a Urinary Tract Infection (UTI) were suspected and were being tested for in the microfluidic chip, the sensitivity for common antimicrobial agents for UTI treatment might include ampicillin (AMP), ciprofloxacin (CIP), and trimethoprim/sulfamethoxazole (SXT), these being three agents that could be tested for and three different paths. The bacteria that might exist within the urine samples from an individual could be any of uropathogenic  E. coli  strains (EC132, EC136, EC137, and EC462). Some prior research has shown that, through antimicrobial resistance profiles of these pathogens that EC132 is resistant to AMP and CIP but not SXT. EC136 is resistant to AMP only. EC137 is sensitive to all the antibiotics tested. EC462 is resistant to AMP and SXT but not CIP. In order to coat sides of the serpentine microchannels  316 , one technique would to have a certain amount of the antibiotic dissolved in sterile water to the serpentine microchannels  316  at different levels. Subsequently, the diluted antibiotic is dried by incubation at a desired temperature and desired time. The original diluted antibiotic has a starting concentration of a predetermined μg/ml concentration. The surface area is sufficiently covered such that, when the biofluid passes thereover, it will interact with reagent. 
     Referring now to  FIG. 6 , there is illustrated a microchannel diagram of the reservoir  330  on the surface of the chip  402 . This is connected by the microchannel  507  from the associated one of the micropumps  504 . After the results in the viewing reservoir  318  have been determined to yield a positive result, for that particular path in the parallel analysis/testing operation, the other of the pair of micropumps  504  is activated and the remaining amount of micro-fluid from the reservoir  418  is transferred to the reservoir  330 . This will be passed through the serpentine microchannel  316  and stored in the reservoir  318 , labeled  602  in  FIG. 6 . This is substantially larger than the reservoir  318  associated with the reservoir  312 . Thus, for this embodiment, the remaining portion of the biofluid from the reservoir  418  will be substantially stored in the reservoir  602 . This will have associated there with a waste microchannel  604  and an outlet microchannel  608  that extends outward from the bottom of the reservoir  602  and up to an opening  610  in the surface of the substrate for interface with the micropump  336 . The micropump  336  is operable, at the next stage of the testing/analysis, to move the contents of the reservoir  602  over to the next reservoir  330  for testing at that next concentration level associated with the next reservoir  330  in the sequence. 
     Referring now to  FIGS. 7A-7D , there is illustrated an example of a valveless MEMS micropump. The micropump includes a body  702  with two pumping chambers  704  and  706 . At the inlet side of each of the chamber  704  and  706  is disposed a conical inlet  710  and  712 , respectively. The conical inlets  710  and  712  are wider at the pump chamber side and narrower at the inlet side thereof. The inlet sides of conical inlet  710  and  712  are connected to respective inlet channel  714  and  716 . The outlet side of the chambers  704  and  706  are interfaced with conical outlets  718  and  720 , respectively, the conical outlets  718  and  720  having a narrower portion at the outlet of the respective pump chamber  704  and  706  and a wider portion at the respective outlet thereof interfacing with respective outlet channels  722  and  724 . The conical inlets  710  and  712  and outlets  718  and  720  are frustro conical in shape. A piezoelectric membrane and actuator  726  is dispose between the two pumping chambers  704  and  706  and is operable to be extended up into one of the chambers  704  and  706  at one time to increase the pressure therein and at the same time extend away from the other of the chambers  704  and  706  to decrease the pressure therein. The operation is then reversed. 
     The piezoelectric membrane and actuator  726  is comprised of a piezoelectric disc  740  on one side of a membrane  742  and a piezoelectric disc  744  on the other side thereof. Each of the piezoelectric discs  740  and  744  are formed by stratifying a layer of use electric material  748  between two layers of conducting material  750 . Piezoelectric material  748  can be made with Piezo Material Lead Zirconate Titanate (PZT-SA), although other piezoelectric materials can be used. The conducting material  60  may be composed of an epoxy such as commercially available EPO-TEK H31 epoxy. The epoxy serves as a glue and a conductor to transmit power to the piezoelectric discs  750 . The piezoelectric discs  750  are secured to the surface of the intermediate layer  748 , so that when a voltage is applied to the membrane  742 , a moment is formed to cause the membrane  742  to deform. 
     The operation of the micropump will be described with reference to  FIG. 7D . At rest, the upper chamber  704  and the lower chamber  706  are separated by a diaphragm pump membrane  742 . The diffuser elements  710 ,  712 ,  718  and  720  are in fluid communication with each respective chamber. Diffuser elements are oriented so that the larger cross-sectional area end of one diffuser element is opposite the smaller cross-sectional area end of the diffuser element on the other side of the chamber. This permits a net pumping action across the chamber when the membrane is deformed. 
     The piezoelectric discs are attached to both the bottom and the top of the membrane. Piezoelectric deformation of the plates is varied by varying the applied voltage so as to excite the membrane with different frequency modes. Piezoelectric deformation of the cooperating plates puts the membrane into motion. Adjustments are made to the applied voltage and, if necessary, the choice of piezoelectric material, so as to optimize the rate of membrane actuation as well as the flow rate. Application of an electrical voltage induces a mechanical stress within the piezoelectric material in the pump membrane  742  in a known manner. The deformation of the pump membrane  742  changes the internal volume of upper chamber  704  and lower chamber  706 . As the volume of the upper chamber  704  decreases, pressure increases in the upper chamber  706  relative to the rest state. During this contraction mode, the overpressure in the chamber causes fluid to flow out the upper chamber  704  through diffuser elements on both sides of the chamber. However, owing to the geometry of the tapered diffuser elements, specifically the smaller cross-sectional area in the chamber end of the left diffuser element relative to the larger cross-sectional area of the right diffuser element, fluid flow out of the left diffuser element is greater than the fluid flow out the right diffuser element. This disparity results in a net pumping of fluid flowing out of the chamber to the left. 
     At the same time, the volume of the lower chamber  706  increases with the deformation of the pump member  742 , resulting in an under pressure in the lower chamber  706  relative to the rest state. During this expansion mode, fluid enters the lower chamber  706  from both the left and the right diffuser elements. Again owing to the relative cross-sectional geometry of the tapered diffuser elements, fluid flow into the lower chamber  706  through the right diffuser element is greater than the fluid drawn into the lower chamber  706  through the left diffuser element. This results in a net fluid flow through the right diffuser element into the chamber, priming the chamber for the pump cycle. 
     Deflection of the membrane  742  in the opposite direction produces the opposite response for each chamber. The volume of the upper chamber  704  is increased. Now in expansion mode, fluid flows into the chamber from both the left and right sides, but the fluid flow from the right diffuser element is greater than the fluid flow from the left diffuser element. This results in a net intake of fluid from the right diffuser element, priming the upper chamber  704  for the pump cycle. Conversely, the lower chamber  706  is now in contraction mode, expelling a greater fluid flow from the lower chamber  706  through the left diffuser element than the right diffuser element. The result is a net fluid flow out of the lower chamber  706  to the left. 
     Referring now to  FIG. 8 , there is illustrated a cross-sectional view of a piezoelectric micropump with check valves. Membrane  802  is disposed within a pump chamber  804  and secured to a body  806 . A piezoelectric disc  808  is disposed beneath the membrane  802  and electrode  810  is disposed below the piezoelectric disc  808 . Deformation of the membrane  802  with the piezoelectric disc at the appropriate frequency will cause a volume of the pumping chamber  804  to change. An inlet valve  810  allows fluid to flow into the chamber  804  and an outlet valve  812  allows fluid to flow out of the chamber  804 . 
     Referring now to  FIG. 9 , there is illustrated a micropump  960  in which a nanofabricated or microfabricated fluid flow pathway is formed between structures. A first reservoir  961  terminates with a first gate valve  966  which permits or restricts fluid flow between the first reservoir  961  and a second reservoir  973 . An electrolytic pump  985  drives a first diaphragm  965  which is communication with the second reservoir  973 , to close the first gate valve  966 , and pulls a second diaphragm  969 , which opens a second gate valve  968  to drive fluid from the second reservoir  973  to a third reservoir  973 . The electrolytic pump  985  is driven by electrowetting of a first membrane  962  on the first gate valve  916  side of the pump. By switching to electrowetting of a second membrane  963 , as depicted in  FIG. 16B , fluid within the third reservoir  973  is emitted from an exit opening  170  by actuation of the second diaphragm  969 . 
     Referring now to  FIG. 10 , there is illustrated a flowchart depicting the overall operation of the system. The process is initiated at a Start block  1002  and then proceeds to a block  1004 , wherein the biofluid sample is loaded. The process enclosed a block  1006 , wherein the biofluid is transferred to the viewing window or the cell counter. The process then flows to a decision block  1008  to determine when the counting operation is done, i.e., when the cells have been discriminated. As noted hereinabove, each of these cells could be associated with, depending on upon the type, a particular affinity label to allow them to be discriminated between within the viewing window. The process then flows to a block  1010  in order to pump the biofluid material to the next phase, that associated with the parallel testing/analysis step. A decision is then made at a block  1012  as to whether this is a positive state, i.e., has any of the biofluid material interacted with a particular reagent to give a positive result. If not, the process is terminated at a block  1014  and, if so, the process flows to a block  1016  in order to capture the biofluid material in a secondary reservoir. Once the path is selected, the appropriate micropump is activated and the biofluid material is pumped to the next reservoir along the secondary path, as indicated by a block  1018 . The process then flows to a block  1022  in order to analyze the results at each secondary reservoir and, if there is a positive result, as indicated by block  1022 , the process is terminated at a block  1024 . If the result is not positive, the process then flows to a block  1026  to determine if that is the last testing reservoir and, if so, the process flows to the terminate block  1024 . If there are more testing/analysis blocks through which to process the biofluid material, the process then flows back to the input of a block  1018  to pump the biofluid serial to the next testing reservoir. 
     Referring now to  FIG. 11 , there is illustrated a flowchart for the loading operation, which is initiated at a block  1101  and then flows to a block  1102  wherein the sample is placed in the well and then to a decision block  1104  to determine if this is a process wherein the biofluid sample is to be mixed with some other diluted product or an affinity label. If it is to be mixed, the process flows to a block  1106  to mix the biofluid sample and, if not, the process bypasses this step. The process then flows to a block  1108  in order to activate the pump and transferred the biofluid material after mixing to the next reservoir in the process. 
     Referring now to  FIG. 12 , there is illustrated a flowchart for the process of the cell counting operation, i.e., the operation at the viewing reservoir. This is initiated at a block  1202  proceeds to a block  1204  in order to transfer the biofluid material to the viewing chamber. The process enclosed a block  1206  in order to view the cells in real time as they pass through the various microchannels and viewing window. The process then flows to a block  1208  in order to count the cells. At this stage, the cells can have various affinity labels associated there with such that the target cells can be viewed and discriminated between based upon the affinity labels associated therewith. If, for example, there were multiple types of bacteria contained within the biofluid sample and each of these types of bacteria had associated therewith different affinity label that clips arrest at a different color, they killed be discriminated between. Additionally, proteins would have a different affinity label than a bacteria and this would also allow discrimination between the two types of cells. The process then flows to a block  1210  to store the transferred biofluid in the reservoir and into a block  1212  to terminate. 
     Referring now to  FIGS. 13A-13C  from their illustrated various configurations for the cell counting operation. In the first embodiment of  FIG. 13A , there are provided a three-part microchannel  1302 , a middle section microchannel  1304  and an outlet microchannel section  1306  the middle section  1304  has a diameter that is slightly larger than the largest cell that could be contained within the biofluid. This allows the cells to be transferred in a more orderly manner. The cell viewing would be performed at this middle section microchannel  1304 . In the embodiment of  FIG. 13B , there are provided three varying diameter middle microchannel sections  1308 ,  1310  and  1312 , each with different diameters to allow different size cells to flow therethrough. This type of embodiment may facilitate some selection in the cells for viewing. In the embodiment of  FIG. 13C , there is illustrated the above disclose embodiment wherein the microchannel  416  empties into the reservoir  418  and the viewing is basically performed upon the cells within the reservoir  418 . 
     Referring now to  FIG. 14  come there is illustrated a flowchart for the parallel cell capture in the first testing/analysis stage. This is initiated at a block  1402  and a process and proceeds to a block  1406  in order to preload all of the cell capture areas having reagent associated there with, such that the portion of the biofluid stored in the reservoir  418  is transferred to the reservoirs associated with the parallel cell capture areas. The process enclosed a block  1408  wherein the pump is activated to fill all of the cell capture wells associated with this stage of testing/analysis. The process then flows to a block  1410  to possibly allow the cells to slowly go through the microchannels in order to interact with the reagent. If so, this requires a certain amount of time and this would result in the micropumps operating at a lower rate to allow sufficient time for the cells to flow through the serpentine microchannels  316  to interface with the particular coating on the surfaces thereof. This basically is the amount of time required for the micropumps to fill up the reservoir  318  associated there with. The length of the serpentine microchannel  316  would determine the amount of time required to fill up the reservoir  318 . Once the reservoir has been filled, as indicated by a block  1412 , then the viewing window in the reservoir  318  is analyzed, as indicated by a block  1414 . The path from the block  1410  to the input of the block  1414  indicates a path by which the micropumps can be run at a higher rate. The process then is terminated at a block  1416 . 
     Referring now to  FIG. 15 , there is illustrated flowchart for the second phase of the analysis, provided that the first phase indicated a positive result for one of the cell capture areas and the associated reagent. This is initiated a block  1502  and then proceeds to a block  1504  to preload all of the secondary cell capture areas with reagent and into a function block  1506  to pump all of the remaining biofluid material from the reservoir  418  into the first reservoir in the secondary reservoirs  330 . This also goes through and incubate step to allow the micropumps to pump at a slower rate to allow the biofluid material to go through the serpentine microchannel  316  at a slower rate before it enters the associated reservoir  318 . When the reservoir  318  is filled, as indicate a by block  1510 , the contents of the reservoir  318  are analyzed at a block  1512 . If the pump can be run at a faster rate, this is provided by a path around the block  1510 . If the result is positive, as indicated by a block  1514 , then the process is terminated at a block  1516 . If not, the process flows from the block  1514  to a block  1518  in order to the next reservoir  330  in the back to the input of the serpentine microchannel  316  and then float the input of the block  1508 . 
     Referring now to  FIG. 16 , there is illustrated a simplified diagrammatic view of the microfluidics chip for processing a plurality of modules. The sample  303  is input to the well  302  and then pumped into the viewing window  306 . A waste microchannel  1602  is provided an interface to the viewing window  306  that is interfaced with a micro valve  1604  to allow air to escape, or any bubbles that may be present, from the viewing window  306 . Additionally, the waste microchannel  1602  could interface with an external vacuum source aid in fluid flow. A cell counter/discriminator  1606  is provided for optically viewing the contents of the viewing window  306 , the output thereof processed via a processor  1608 . The outlet of the viewing window  306  is interfaced with a manifold microchannel  1610  through a connecting channel  1612 . At this point, the micro valve  1604  is closed such that the biofluid contained within the viewing window  306  and the interfaced with microchannel manifold  1610  to allow fluid to be pump therefrom to a plurality of distribution paths along distribution microchannels  1614 . It may be that pump  304  would need to be activated in order to reduce the pressure at the upper end of the viewing channel  306  or, alternately, a microchannel  1618  interfaced with a micro valve  1620  could be provided to, when open, relieve the pressure in the upper end of the viewing window  306  to allow biofluid to be pumped therefrom to the microchannel manifold  1610 . 
     Each of the distribution microchannels  1614  is interfaced with a separate module via an associated distribution pump  1624  to interface with and associated one of modules  1625 , labeled A-Z, for example. There can be any number of modules provided. However, each module  1625  has associated there with a finite capacity and, therefore, the number of modules  1625  that can be interfaced to the viewing window  306  is a function of the volume of biofluid contained therein and the capacity of the reservoirs of each of the individual modules  1625 , each of the individual modules  1625  potentially having a different capacity, depending upon the configuration thereof. However, selecting among the various distribution pump  1624  can allow desired tests to be done with the available biofluid contained within the viewing window  306 . 
     Referring now to  FIG. 17  there is illustrated a diagrammatic view of one of the modules  1625  associated with the parallel testing configuration, wherein biofluid is loaded into a plurality of testing reservoirs. The distribution pump  1624  associated there with transfers fluid from the distribution microchannels  1614  to an intermediate microchannel manifold  1702  which is then interface with a plurality of testing reservoirs  312 , as described hereinabove. Each of these testing reservoirs has a serpentine microchannel  312  and a viewing window  318  associated there with. As described hereinabove, each of these testing reservoirs can have a different volume and a different configuration mechanically and can be associated with a different test. They can each have a particular coating of reagent, such as an antibiotic, to interact with the biofluid for testing purposes to determine if there is any reaction of the biofluid in the cells contained therein to the material coated on the sides of the serpentine microchannels  316 . In the operation of this particular module  1625 , all of these testing reservoirs  312  are associated with different reagents and will be loaded in parallel. For this embodiment, will be desirable for each of the reservoir  312  to have the same volume. If, however, they had different volumetric capacities, it would be necessary to have some type of waste gate with a micro valve to allow all of the viewing windows  318  to achieve full capacity. 
     Referring now to  FIG. 18 , there is illustrated a diagrammatic view of the serially configured wherein a plurality of testing reservoirs  330  our arranged in a series configuration. In this configuration, the associated distribution pump  1625  will transfer biofluid from the microchannel manifold  1610  through the distribution microchannels  1614  to the first of the testing reservoirs  330 . The biofluid will be contained within the viewing chamber  318  and, as noted hereinabove, there will be possible he some type of waste microchannel associated micro valve to allow air/bubbles to escape during filling of the viewing window  318 . Thereafter, a second serial pump  1706  is activated to transfer the contents of the viewing window  318  to a second testing reservoir  330  in the associated serpentine microchannel  316  and viewing window  318 . In this transfer, there may be required a relief microchannel (not shown) at the inlet end thereof to reduce the pressure therein during the pumping operation. This will continue until all of the tests have been done. Each of the serpentine microchannels  316  associated with each of the testing reservoirs  330  will have a graduated increase in the particular reagent to determine the dosage, in this example. It may be that, upon being exposed to the dosage of the reagent in the first testing reservoir  330  that cellular material in the biofluid is somewhat affected by the reagent, i.e., the antibiotic, for example. By moving to a higher concentration of the reagent in the next sequential testing reservoir  330 , this could be accounted for in the overall analysis. It may be that the actual concentration in the next sequential testing chamber  330  is not an exact incremental increase in the reagent. For example, if it was desired to expose the biofluid to reagent increments of 10%, 20%, 30%, etc. in 10% increments, it may be that the first testing chamber  330  has a concentration of 10% and then the second testing chamber has a concentration of possibly 16%, accounting for the fact that the accumulated effect of passing through the 10% testing chamber  330  and the 16% testing chamber  330  effectively provides a 20% accumulated exposure in the second testing chamber  330  and so on. 
     Referring now to  FIG. 19 , there is illustrated a diagrammatic view of a configuration for providing parallel loading of the serial configuration for the incremental testing. This is similar to the embodiment of  FIG. 17 , except that the testing chambers  330  are all interfaced with the associated distribution pump  1625  through a microchannel manifold  1902  in a parallel configuration, such that they are all loaded at the same time, with each having a different concentration of reagent associated there with. In this configuration, however, since all of the testing chambers  330  will be loaded in parallel, there are required to be a sufficient volume of biofluid contained within the viewing window  306  initially to facilitate complete filling of each of the associated viewing windows  318 . 
     Referring now to  FIGS. 20A-20B  come there is illustrated a diagrammatic view of chemostat, wherein the associated distribution pump  1625  transfers biofluid from the distribution microchannel  1614  to a chemostat  2002 . The details of the chemostat  2002  are illustrated in  FIG. 20B . A main microchannel  2004  is interfaced on one and thereof with the output of the distribution pump  1625  associated there with, with the other end of the microchannel  2004  interfaced with a waste gate via a micro valve (not shown). There are a plurality of cell storage microchannels  2006  connected between one surface of the main microchannel  2004  and a waste microchannel  2008 . Each of these cell storage microchannels  2006  associated there with a filter  2010  disposed at the end thereof proximate to the waste microchannel  2008 . Each of the cell storage microchannels  2006  has a size that will receive a particular target cell having a particular dimension, such that the target cell will flow into the cell storage microchannel and cells of smaller size will pass through the associated filter  2010 , which filter  2010  is a microchannel with a diameter that is smaller than that of the target cell. This waste material will flow out through the waste gate or micro valve (not shown) associated with the waste microchannel  2008 . By maintaining a pressure differential between the main microchannel  2004  and the waste microchannel  2008 , the target cells will be stored within the cell storage channels  2006 . Larger cells than the target cells in the main microchannel  2004  will bypass the cell storage microchannels  2006  and pass out of the waste gate associated with the main microchannel  2004 , keeping in mind that there is required to be a lower pressure within the waste microchannel  2008  as compared to the main microchannel  2004 . 
     Referring now to  FIG. 21 , there is illustrated an embodiment of the microfluidic chip utilizing micro valves as opposed to intermediate micropumps. In this embodiment, there are illustrated a plurality of input wells  2102  for interfacing with an initial micropump  2104  to pump fluid through a viewing window  2106  to a first reservoir  2108 . Having multiple wells  2102  allows multiple samples to be input through the viewing window  2106  or to actually mix two different materials together for flowing through the viewing window  2106  to the reservoir  2108 . The waste gate  2110  can be provided at the reservoir connected thereto via a waste microchannel  2112  to allow air/bubbles to escape. A micropump  2114  is operable to pump fluid from the reservoir  2108  to a main microchannel manifold  2116 . During this pumping operation, some type of pressure relief is required which can either be provided via one of the pumps  2104  being activated or a relief micro valve  2118  Interface with the input end of the viewing window  2106  through a relief microchannel  2120 . 
     Interfaced with the main microchannel manifold  2116  is a plurality of distribution micro valves  2124 . These distribution micro valves  2124  can be interfaced with various modules, as described above herein with respect to  FIGS. 17-20A /B. The only difference is that the associated distribution pump  1624  has been replaced by a distribution valve  2124 . Additionally, each of the parallel loaded testing reservoirs  312  can be individually associated with one of the distribution valves  2124  to selectively certain ones thereof for testing. Since each one of these testing reservoirs  312 , after selection, is required to be completely filled, by allowing individual selection of the testing reservoirs  312 , certain ones thereof can be eliminated. It may be that, in pre-analyzing the biofluid sample, it can be predetermined that certain ones of the associated reagents in the reservoir  312  are not required the testing/analysis step and can therefore be eliminated from the step of filling. This is opposed to the embodiment of FIG. 17 , wherein all of the testing reservoirs  312  are complete the filled. 
     Referring now to  FIGS. 22A-22B , there is illustrated cross-sectional views of a micro valve in an open and a closed position. The substrate  402  has cover plate  440  disposed on top thereof. There are provided to microchannels  2202  and  2204  that are to be connected together with the micro valve. The microchannel  2202  is interfaced with a hole  2006  to the surface of the cover plate  440  to an opening  2208 . The microchannel  2204  is interfaced to a hole  2210  to an opening  2212  in the cover plate  440 . The micro valve has a fixed body  2214  with a membrane  2216  disposed on the surface there above to define a pumping chamber  2218 . The pumping chamber  2218  has a hole  2220  interfacing the pumping chamber  2218  with the opening  2208  on the cover plate  440 . Similarly, the hole  2212  is interfaced to the pumping chamber  2218  through a hole  2222 . The membrane  2216  is operable to reciprocate away from the surface of the fixed body  2214  exposing the top of the hole  2220  in the pumping chamber  2218  to allow fluid to flow through the pumping chamber  2218  and down through the opening  2222  through the cover plate  440  and through to the microchannel  2204 . In the closed position, the membrane  2216  is forced down against the upper end of the hole  2220 . A pneumatic cavity  2230  is disposed above the membrane  2216  in a housing  2232  and interfaces with a pneumatic source through a hose  2234 . Thus, by drawing a vacuum in the pneumatic cavity  2230 , the membrane  2216  will be pulled away from the hole  2220  to allow fluid to flow and, when pressurized air is forced into the pneumatic cavity  2230 , and the membrane  2216  is forced down to the surface of the fixed body  2214  to seal the opening  2224  in a closed position. 
     Referring now to  FIG. 23 , there is illustrated a process flow for preparing the biologic sample for the microfluidic chip  102 . The preparation of the biologic sample can take many forms. In this example, the raw biologic sample can be preprocessed, depending upon the type of sample that is being considered. For example, if blood is being tested, the Complete Blood Count (CBC) can be determined, as well as the White Blood Cell Count (WBC), the liver functions and the kidney functions. For urinalysis, the sample can be prepared for testing for WBC&#39;s and nitrates, as well as proteins and Bilirubin. There are many well-known processes for preparing biologic samples prior to testing. Once the biologic sample has been prepared a, affinity labels are attached thereto. Typically, there will be a vial  2302  provided with the biologic sample that is mixed with affinity labels in a vial  2304  resulting in the vial  2304  containing a labeled sample. These labels are sometimes referred to as “affinity labels” or “microspheres.” These functional polymeric microspheres typically have a diameter of less than 5 μm and have been developed for use with immunological methods. The reagents were initially used as visual markers to identify specific cell types and analyze the distribution of cell surface antigens by scanning electron microscopy. They have also been used, due to their inherent properties, two separate labeled from unlabeled cells by techniques such as centrifugation, a electrophoresis, magnetic chromatography and fluorescence cell sorting. The cells contained within the biologic sample are basically cells bearing defined antigens or receptors, ligands which bind with a high degree of selectivity an affinity to these cell surface sites. The microspheres interact with the specific ligand, which can allow for separation based upon the characteristic properties of the microspheres. This allows for displaying of these labeled cells with the target receptor or antigen with antibiotics or other ligands directly or indirectly bound to the microspheres. Specific types of microspheres or affinity labels can be the type that will fluoresce at a particular wavelength. Thus, specific cells can be identified the optical techniques to identify target cells or differentiate between various types of, for example, bacterial cells and proteins, etc. This labeled sample is then disposed within the well  302  on the microfluidic chip  102  for later processing. 
     Referring now to  FIG. 24 , there is illustrated a side cross-sectional view of an RT-lamp. The RT-lamp is a Reverse Transcription Loop-mediated isothermal Amplification device, which is an a technique for the amplification of RNA. This combines the advantages of the reverse transcription without of the LAMP technique. The LAMP technique is a single to technique for the application of DNA. This technique is an isothermal nucleic acid application technique, in which a chain reaction is carried out at a constant temperature and does not require a thermal cycler. The target sequences animal five at a constant temperature using either two or three sets of primers and polymerase with high strand displacement activity in addition to a replication activity. The addition of the reverse transcription phase allows for the detection of RNA and provides a one-step nucleic acid amplification method that is used to diagnose infectious diseases caused by bacteria or viruses. 
       FIG. 24  illustrates an example in which a multimode instrument  2401  is coupled to a smartphone  2402 . The smartphone  2402  includes an LED  2404  and a camera  2406 . The camera  2406  includes an image sensor, such as a CCD. The instrument  2400  includes a sample chamber  2408  for receiving an optical assay medium. The optical assay medium comprises the labeled biologic sample disposed within the viewing window  306  on the microfluidic chip  102 . The sample chamber  2408  may include a door  2410  that prevents stray light from entering 
     The optical assay medium is positioned over a detection head  2412  in the sample chamber  2408 . The instrument  2400  includes an optical output path for receiving an optical output from the optical assay medium in the sample chamber  2408  via the detection head  2412 . The optical output path may include a multimode fiber  2414  that directs light from the detection head  2412  to a cylindrical lens  2416 . The optical output path may further include a wavelength-dispersive element, such as a diffraction grating  2418 , that is configured to disperse the optical output into spatially-separated wavelength components. The optical output path may also include other optical components, such as collimating lenses, filters, and polarizers. 
     The instrument  2401  can include a mount for removably mounting the smartphone  2402  in a working position such that the camera  2406  is optically coupled to the optical path, for example, in a predetermined position relative to the diffraction grating  2418 . In this working position, the camera  2406  can receive at least a portion of the dispersed optical output such that different locations are received at different locations on the image sensor. 
     The instrument  2401  may also include an input optical path for directing light from a light source to the optical assay medium in the sample chamber  2408 , for example, through the detection head  2412 . In some instances, the LED  2404  on the smartphone  2402  could be used as the light source. To use the LED  2404  as the light source, the input optical path may include a collimating lens  2420  that receives light from the LED  2404  when the smartphone  2402  is mounted to the instrument  2400  in the working position. The input optical path may further include a multimode fiber  2422  that directs the light from the collimating lens  2420  to the detection head  2412 . The input optical path may also include other optical components, such as collimating lenses, filters, and polarizers. 
     The instrument  2400  may also include an additional input optical path that directs light form an internal light source, such as a laser  2424 , to the optical assay medium in the sample chamber  2408 . The additional input optical path may include a multimode optical fiber  2426 , as well as collimating lenses, filters, polarizers, or other optical components. 
     Referring now to  FIG. 25 , there is illustrated the view of the RT-lamp  2401  with a microfluidics chip  102  disposed within the sample chamber  2408 . 
     Referring now to  FIG. 26 , there is illustrated a side view of the smart phone  2402  interfaced with the microfluidic chip  102  four imaging the surface thereof, which is illustrated in a window view in  FIG. 27 . This window view illustrates the viewing window as a box  2702  in which the image of the microfluidic chip  102  is displayed. The application automatically recognizes various markers  2704 ,  2706  and  2708  1 three corners thereof. This will allow orientation of the window with respect to the application. A box  2710  shown in phantom dashes will be oriented by the application running on the smart phone  2402 . Once the box has been oriented visually about the image of the microfluidic chip  102 , then processing can proceed. The processing is basically focusing upon the chip to gain the best optical image of the target sites. The target sites are storage reservoirs  312  and  330 , for example. Each of these will have a viewing well  318  associated there with and these viewing wells  318  will have, in one example, a process biologic sample having affinity labels associated there with that fluoresce. By recognizing the florescence, the presence of the cell can be determined. The lack of florescence indicates that the cell, a bacteria for example, has been destroyed. This can be a positive test. By examining at each stage of the testing process the chip, a determination can be made as to results in essentially real time. This will be described in more detail hereinbelow. Once the image is believed to be in focus, the user can actually take the picture or the application can automatically determine that the focus is adequate and take that. This is very similar to character recognition techniques that are utilized in recognizing faces in camera images received by the phone. 
     Referring now to  FIGS. 28A-28H , there are illustrated various images of the microfluidic chip  102  at different stages, this view being a diagrammatic view for simplicity. In this view, there is provided the sample well  2802  which then feeds into the viewing well  2804 . As described hereinabove, there are multiple pumps that allow fluid to be moved from the sample well  2802  over to the viewing well  2804  and these are not shown for simplicity purposes. 
     There is provided a multiplexer  2802  which represents the micropumps/valves described hereinabove. The multiplexer  2008  may be associated with one bank  2808  of reservoirs  2802  for the parallel processing stage. These reservoirs  2012  correspond to the reservoirs  312 . This requires that the multiplexer  2806  distribute fluid to a microchannel manifold  2810  and one testing phase. The multiplexer  2006  also is connected via a plurality of microchannels to a bank of reservoirs associated with the serial processing stage to selectively distribute fluid to one of the strings in a second testing phase. This bank of reservoirs includes the reservoirs  330  described hereinabove. Each of these reservoirs  330  is arranged in series such that each has a valve or pump disposed there between. The multiplexer  2806  also interfaces with a bank  2830  of reservoirs, these, in this example, associated with the serial testing/analysis stage and having reservoirs  330  associated there with. In this example, there are provided five test reservoirs in the bank  2808 , wherein each of these test reservoirs has associated there with one serial string of test reservoirs  330  in the bank  2814  and one parallel loaded string of reservoirs  330  in the bank  2820 . Additionally, there is a separate testing reservoir  2824  which could correspond to the cell storage area utilizing a chemostat described hereinabove, which is interfaced with multiplexer  2806  through a microchannel  2826 . 
     Referring now to  FIGS. 28B-28H , there are illustrated various stages of the loading and analysis.  FIG. 28B  illustrates the first step in the process wherein the biologic sample is loaded into the viewing window  2004 . That the step in the process, the microfluidic chip is disposed within the RT lamp  2401  and analyzed to determine the number of cells and the type of cells. If, for example, a certain bacteria were being tested for on this particular microfluidic chip  102 , the lack of bacteria cells, as indicated by the particular affinity labels that would be attached to these particular bacteria cells, would indicate that further testing is not required. However, if the correct cells are labeled and the number of cells is at an appropriate level for testing, then the next step of the process is taken. 
       FIG. 28C  illustrates a next step of the process wherein a portion of the contents of the viewing well  2804  are transferred to all of the reservoirs  2812  in the bank  2808 , there being five reservoirs  2812  disposed therein, it being noted that there could be more reservoirs  2012  provided on the microfluidic chip  102 . There will be a certain amount of time required for the pump associated with the multiplexer  2808  to actually move the desire portion of the biologic sample through the manifold  2810  to the reservoirs  2812 . As noted hereinabove, each of the reservoirs  2812  corresponds to the reservoirs  312 , each having a serpentine microchannel  316  and a viewing reservoir  318  associated there with. The micro pumps associated with the multiplexer  2806  in communication with the very small widths of the microchannels can require this process to take upwards of 10 or 20 minutes. After this period of time, the microfluidic chip  102  can be imaged to determine if the cells have been destroyed by the coating on the surfaces of the serpentine channel  316 . (It should be noted that the viewing well  318  could also be coated). If the cells are destroyed, this indicates that the reagent that coats the walls of the microchannel associated with the reservoirs  312  reacted in a manner indicating self-destruction. However, any visual indication in the viewing wells that can be a vehicle for discrimination between interaction with the particular reagent coating the walls of the serpentine microchannels  316  will provide the ability for a decision to be made as to which reagent is required for further testing. 
       FIG. 28D  illustrates the next phase of operation, which is the phase in which the dosage level is determined. In the example above, the middle reservoir in the bank  2808  provided a trigger indication that triggered a decision to then test for dosage in the middle string within the bank  2814 . This will require a multiplexer  2806  to only transfer the remaining portion of the biologic sample from the viewing reservoir  2804  into this particular string. As described hereinabove, this process will involve first passing of fluid to the first reservoir  330 , which will take a certain amount of time to actually pump the biologic sample through the microchannels into the viewing window  318 . This can be a multiphase process, which requires viewing at each stage. In this particular example, the third stage of testing in this middle string in the bank of reservoirs  2814  resulted in a perceivable result, i.e., a lack of florescence, for example. At this point, the image will actually show the perceivable result in both the bank  2808  and in the bank  2814 . Thus, in the three phases of testing, the particular cells have been a defined, an indication has been provided as to which of multiple reagents that could possibly provide the desired therapeutic results would be the best choice for the patient and then the third phase of testing provides the actual dosage of that determine reagent. It may be that for ten individuals that had exactly the same symptoms and processed a similarly processed biologic sample for testing in the same way with the microfluidic chip and the RT-lamp  2401  came up with different results. Each individual&#39;s particular physiology can vary and, as such, the results could differ. In a typical medical environment, the particular reagent of choice or drug of choice is determined by an individual based upon various criteria. Since the medical professional does not have the test directly in front of them, they might just prescribe, for example, a broad based antibiotic. They might follow that up with testing of a biologic sample in a lab, which could take a number of days just to determine exactly what bacteria is present and what would be the best antibiotic to use in order to attack this particular bacteria. Of course, the broad-spectrum antibiotic might have worked by the time the test results come back. If not, these results might be useful to the medical professional. However, these tests seldom if ever actually focus in on the dosage that would be preferable for a particular individual. If even the particular antibiotic could be identified which was specific to that particular bacteria tested for and found be present in the biologic sample, the dosage prescribed is typically a medium or high dosage, depending upon the criteria that the medical professional utilizes. However, the medical professional typically generalizes the physiology of any individual and maybe filters that based upon age, gender, etc. However, the individual physiology is not taken into account. 
     With use of the present microfluidic chip  102 , the entire testing process can be performed at the Point of Care (POC) in a relatively short amount of time. The result is not only the identification of the best reagent to use but also the dosage. This is all accomplished with a very small amount of biologic sample. 
       FIG. 28E , there is illustrated a potential further processing that can be provided. In this embodiment, the bank  2820  can have a different modification of the antibiotic that was determined from the test associated with the bank  2808 . This modification could be associated with the pH of the antibiotic, wherein it has been determined with respect to some antibiotics that the pH of the antibiotic can affect the efficacy thereof. In this example, it can be seen that the third reservoir with respect to dosage is the one that is selected in the bank  2814  but in the bank  2820 , is the lowest dosage. Thus, the multiplexer  2806  needs to first test the bank  2814  and then test the bank  2820 . However, it should be understood that both the bank  2814  and the bank  2020  could be identical, either serially loaded or parallel loaded, the commonality being that they have a gradually increasing dose of antibiotics that can be tested for. 
       FIGS. 28F-28G , there are illustrated two additional examples of two different patients with substantially the same symptoms and utilizing substantially the same process for preparing the biologic sample. With respect to  FIG. 28F , the fifth reservoir and the antibiotic associated there with exhibited the highest efficacy at the highest dose as to destroying the particular bacteria, in the example of the bacteria. The associated dosage determined from testing the biologic sample in the bank  2814  was considered to be the second level of dosage. In the bank  2020 , the third level of dosage was considered to be the lowest dose. With respect to  FIG. 28G , the first reservoir and the antibiotic associated there with was considered to have the highest efficacy with respect to dealing with the particular bacteria and it was the lowest dose in that case when tested in the bank  2814 , as compared to the fourth level dosage in the bank  2820 . It can be seen thus that different patients will have different “fingerprints” associated with the testing of the same biologic sample repaired and substantially same way. 
       FIG. 28H , there is illustrated an alternate embodiment wherein the test performed at the bank  2808  resulted in a slight ambiguity in that the bacteria were killed in two other reservoirs. In this case, the indication would be that either of these antibiotics would work against this particular strain of bacteria. Thus, the next phase the test would require the multiplexer  2808  to distribute the contents of the reservoir  2804  through the microchannels to actually two different strings. Thus, for this type of test to be carried out, it is important that there be sufficient volume in the viewing window  2804 , i.e., sufficient amount of biofluid introduced to the well  2802 , in order to fill both of these reservoirs and allow the testing to progress down to the highest dosage level in either or both of the banks  2814  and  2820 . The results of this test show that, for the rightmost reservoir in the bank  2808  having been determined to be effective at the highest dose, the next of the last dosage was required in order to achieve the desired results, whereas the next to the left reservoir in the bank  2808  having been determined to be effective at the highest dose required only the smallest dose to achieve the results. Therefore, this test shows that, although two antibiotics would work, one would actually work with the lower dose. 
     It should also be understood that, in addition to the test being different for the same strain of bacteria in a biologic prepared men substantially the same way, it should also be understood that this particular set of results could be different for different strains of the same bacteria. It may be that, for one strain, one antibiotic would work at a particular dose and, for another strain of the same bacteria, a different antibiotic work or just a different dose of the same antibiotic. The microfluidic chip described and disclosed in the present disclosure allows this determination to be made utilizing a single sample in a parallel/serial testing method at the POC wherein the first step or phase of selection is made among a plurality of potential antibiotics that could arguably target different bacteria and, once a determination is made at the first phase, then the next and serial decision is made to determine dosage at a second phase. 
     Referring now to  FIG. 29 , there is illustrated a flowchart depicting the overall analysis process. The process is initiated at a block  2902  and then proceeds to a block  2904  wherein the biologic sample is prepared. As described hereinabove, this preparation involves labeling the cells within the biologic sample so that they can be discriminated between or identified. It may be that there are a number of different types of cells such as bacteria of different strains and types, proteins, etc. Different affinity labels can be applied such that multiple cells of different types can be identified. The process then flows to a block  2906  wherein the biologic sample is placed into the sample well and then passed on to the viewing well. At this point, the microfluidic chip is placed into the RT-lamp and optically analyzed, as indicated by process block  2908 . It is at this point in the testing phase that the identification process will identify the potential target cells. Since each of the microfluidic chips has a finite number of reservoirs associated there with for the purpose of testing, the coating is applied to these particular reservoirs for the specific antibiotics or reagents to be tested may not be useful for testing the particular cellular structures that have been identified at this step in the process. However, it should be understood that the number of different banks of testing reservoirs that can be provided on a particular microfluidic chip can be expandable and the could actually be provided for multiple different types of reagents. For example, one set of testing banks may be associated with UTI and another associated with streptococcal bacteria. Recognizing these at this step in utilizing them with a microfluidic chip that can test for both types of bacteria will allow the particular biologic sample, which is quite small, to be routed to the appropriate reservoirs for testing for that specific identify bacteria. 
     The decision to proceed is determined at a decision block  2910  and, if testing can proceed with the current microfluidic chip, the process proceeds to a block  2911  to select the particular test that are to be performed. The process then proceeds to sequence through the tests, as indicated by a block  2914 . This sequencing sequences through the various phases, with the initial test being selected first, as indicated by block  2916 . In the above examples, this is the first parallel phase to determine which among several reagents is most effective against the particular cellular structure of interest. The process then proceeds to a block  2918  in order to analyze the results of this initial test and then to a decision block  2920  to determine if more tests are required or if this is the only test. If the test is negative at this stage and none of the reagents provides any effectiveness indication, the process is terminated or, if this is the last test, the process is terminated. The process, if continued, then selects the next test in the sequence and proceeds back to the input of the block  2918  to continue sequencing through the tests. 
     Referring now to  FIG. 30 , there is illustrated a flowchart for the testing process. This is initiated at a block  3002  and then proceeds to a block  3004  two first pump a portion of the biologic sample stored in the viewing window through to the parallel reservoirs and load all of the parallel reservoirs for testing/analysis. This may take upwards of 10 or 20 minutes, due to the fact that the micropumps utilized are relatively slow and the diameter of the microchannels is small, thus restricting high flow rates. The process then flows to decision block  3006  to determine if there is been any positive result, i.e., is there any indication that any of the reagents provide an effectiveness indication, either through some color change or the lack of color indicating the destruction of the cells. If there is no result, then the process is terminated and the process flows to a function block  3008  to select the next test path that is associated with the antibiotic having been tested as being effective in the first phase of operation/testing. A process block  3010  and indicates that a graded dosage test path is selected, either the one for loading parallel or the one or loading serially. It should be understood that the parallel loaded graded dosage test path requires all of the reservoirs to be completely filled from the reservoir associated with the viewing window. The serial path, by comparison, allows all of the contents of the viewing window in the reservoir associated there with to be disposed in each reservoir and then sequentially transferred to the next reservoir down the chain and at the higher dosage. However, it should be understood that the system can be configured such that the first reservoir at the lowest dosage is loaded with only a portion of the contents of the viewing window and the reservoir associated there with, analyzed and then a micro valve gate opened to allow the micropumps for pumping fluid to the serial path to operate to continue pushing more biofluid through the first reservoir, thus filling the second and reservoir and so on. In this process, sufficient biofluid must be contained within the viewing window and the reservoir associated there with in order to allow for filling of all of the reservoirs down to the highest dosage rate associated with that serial string. 
     In the process, the serial string will first select the lowest graded dose reservoir and a process block  3012  and then pump biofluid to the first reservoir and a process block  3014 , analyze the results a process block  3016 , understanding that it could take  10  to  20  minutes to fill each reservoir. A determination is made at a decision block  3018  as to whether there is a positive result, i.e., was there and an effectiveness determination made at this point, and, if so proceed to a decision block  3020  to determine if there are any higher concentrations to be tested for. If so, the next reservoir selected by opening gate or activating a micropump, as indicated by a process block  3022 , and the proceed back to the process block  3014  in order to pump to this reservoir. 
     In the parallel process, a process block  3026  indicates an operation wherein the micropump pumps sufficient biofluid material to the parallel rated reservoirs to fill all of the reservoirs and into a process block  3028  in order to analyze the results. 
     It will be appreciated by those skilled in the art having the benefit of this disclosure of a System and Method for Determining Efficacy and Dosage Using Parallel/Serial Dual Microfluidic Chip. It should be understood that the drawings and detailed description herein are to be regarded in an illustrative rather than a restrictive manner, and are not intended to be limiting to the particular forms and examples disclosed. On the contrary, included are any further modifications, changes, rearrangements, substitutions, alternatives, design choices, and embodiments apparent to those of ordinary skill in the art, without departing from the spirit and scope hereof, as defined by the following claims. Thus, it is intended that the following claims be interpreted to embrace all such further modifications, changes, rearrangements, substitutions, alternatives, design choices, and embodiments.