Patent Publication Number: US-6218193-B1

Title: Precision small volume fluid processing apparatus and method

Description:
This application is a divisional application of U.S. patent application Ser. No. 08/531,215, filed Sep. 19, 1995, and entitled “Precision Small Volume Fluid Processing Apparatus and Method,” now U.S. Pat. No 5,785,926. 
    
    
     TECHNICAL FIELD 
     The invention relates to methods and apparatus for precisely handling small volumes of fluids. More specifically, the invention relates to methods and apparatus for aliquoting and assaying biological fluid samples. 
     BACKGROUND OF THE INVENTION 
     Diagnostic and other biological assays often require systems for metering dispensing and mixing reagents with sample fluids. The sample fluids may include, for example, patient samples, blood samples, or minute quantities of deoxygenated rybo nucleic acid (hereinafter “DNA”) sequences in a buffer fluid. Both manual and automated systems have been available for aliquoting the fluid samples, and assaying the samples with one or more reagents. Manual systems have historically included the glass capillary pipette, the micro pipette, precision syringes and weighing equipment. A variety of biological assays have been and continue to be conducted with manual equipment of the type described. 
     Relatively sophisticated microbiological assays including micro-enzyme linked immunosorbent sandwich assays (hereinafter “ELISA”) can be satisfactorily, if tediously performed manually. The demands of modern antibody/antigen matching, histocompatibility typing, paternity testing, etc. on a vast scale has precipitated the development of various automated assay equipment to more quickly process large numbers of patient samples with various reagents. It is apparent that in order to perform a multiplicity of assays with a single patient sample, the amount of sample must be relatively large, or a small sample must be aliquoted into smaller divisions. 
     Recent advances in microbiology have provided the biotechnologist with increasingly sophisticated tools for examining genetic material. Restriction enzyme digestion and polymerase chain reaction (hereafter “RED and PCR respectively”) have provided geneticists with multiple DNA segments from a single sample for subsequent assaying. All of these advances have increased the need for sample handling and processing techniques which are beyond the ability of the heretofore manual pipetting and other standard laboratory techniques. As a result, the industry has proceeded with the development of highly automated equipment which can rapidly and repeatably handle relatively small quantities of patient samples. 
     The undertaking of the Human Genome Project exceeds the limits of current fluid sample handling and processing technology. The Human Genome Project is an attempt to map the entire human genetic code, nucleotide by nucleotide. The PCR and RED techniques presently available will therefore produce an extremely large numbers of nucleotide segments which must be assayed in a variety of different ways. In addition, current methods for producing the nucleotide segments are extremely expensive requiring the very wise use of the resultant sample. It is currently calculated that without further advancement in the state of the art, the cost of producing sufficient samples for laboratories around the world will be prohibitive without the development of techniques for handling much smaller samples and reagent volumes. 
     Therefore, a need exists for a high-precision, small volume fluid processing system which can aliquot and dispense fluid samples in extremely small volumes, react the samples with small quantities of reagents, and perform all of the other steps which may be necessary in a conventional assay. The system should also preferably be relatively highly automated so that the incidence of human error is reduced. 
     SUMMARY OF THE INVENTION 
     It is therefore an object of the present invention to provide a high precision, small volume fluid processing system which can precisely aliquot small volumes of a sample fluid. 
     It is further object of the invention to provide a high-precision, small volume fluid processing system which can mix small aliquots of sample fluid with various discreet fluid reagents. 
     It is a further object of the present invention to achieve the above objects in a system which automatically aliquots the fluid sample, introduces appropriate reagents, mixes the sample and reagents, and incubates the same in preparation for gel electro phoresis. 
     It is yet a further object of the present invention to achieve the above objects in a system which can precisely and repeatably handle fluid volumes as small as 0.1 μl. 
     The invention achieves the objects, and other objects and advantages which will become apparent from the description which follows by providing a small volume fluid processing system employing at least one small volume capillary tube. A precision linear actuator connected to a computer controlled motor acts as a pneumatic piston to precisely inspire and expire one or more fluids into or out of the capillary in a predetermined sequence. 
     In a preferred embodiment, the fluid processing system can include a sample fluid station for containing an initial volume of sample material in a buffer solution or water, a reagent fluid dispensing device, monitoring equipment for determining the position of a fluid segment in the capillary tube and a mechanism for precisely positioning the fluid sample handling device in a reference plane with respect to the reagent fluid dispensing device in the sample fluid station. The sample fluid handling device can intake a precise volume of sample fluid from the sample fluid station, position itself adjacent to the reagent fluid dispensing device which can dispense an appropriate reagent into an open end of the capillary tube. The monitoring device can provide information to a computer or other management system to either advance or retard the precision linear actuator so as to move the fluids appropriately in the capillary tube. If two or more fluids have been received in the capillary tube, the linear actuator can be advanced and retarded with differential velocities so as to mix the fluids in the capillary tube. Very small volumes of fluids, as little as 0.1 μl can be handles with an accuracy of ±0.01 μl and similar repeatability if the precision linear actuator is driven by a computer controlled motor rotatably connected to a precision lead screw. The system described above may optionally contain a heating and cooling system for incubating the capillary tube in a controlled manner. The heating system can include a thin layer of highly resistive, transparent material on the outside of the tube which can be electrically excited so that a heat generating current flows therethrough. A fan can be used to cool the capillary tube while it is heated or afterwards to maintain a desired temperature or quickly cool the tube. The monitoring device for determining the position of a fluid segment in a capillary tube can be of the optical type including a light emitter/detector pair or array positioned in proximity to the capillary tube. 
     The sample fluid handling device can form a bubble or droplet of sample, reagent or other fluid on the open end of the tube having a known volume. By positioning a second capillary tube in alignment with and adjacent to the first capillary tube the droplet can be transferred from the first tube to the second tube by advancing the tubes towards one another until the droplet touches the second tube. Capillary action draws the droplet into the second tube in a repeatable manner. In this way, very small quantities of fluid can be transferred from one capillary to another. In an aliquoting method employing the processing system, the sample fluid handling device can inspirate aliquots of fluid sample separated by air gaps therebetween for a more precise dispensing of the aliquots into separate receiving capillary tubes. 
    
    
     BRIEF DESCRIPTION OF THE DRAWINGS 
     FIG. 1 is an isometric, environmental view of a high-precision, small volume fluid processing system of the present invention. 
     FIG. 2 is a side elevational view of the system shown in FIG.  1 . 
     FIG. 3 is an enlarged, partial side elevational view of a capillary tube of the sample fluid handling device approaching a sample fluid station. 
     FIG. 4 is an enlarged, partial side elevational view similar to FIG. 3 showing the capillary tube inspiring a precise volume of sample fluid. 
     FIG. 5 is an enlarged, partial elevational view similar to FIG. 4 showing the sample fluid handling device being retracted from the sample fluid station. 
     FIG. 6 is an enlarged, partial elevational view of a reagent fluid dispensing device projecting reagent fluid droplets into the capillary tube. 
     FIG. 7 is an enlarged, elevational view of the capillary tube undergoing a mixing action. 
     FIG. 8 is a view similar to FIG. 7 showing the results of the mixing action. 
     FIG. 9 is a schematic representation of a piezo electric reagent dispenser. 
     FIG. 10 is an enlarged, partial sectional view of the circle area of FIG.  9 . 
     FIG. 11 is a schematic representation of process steps employed by the system of the present invention. 
     FIG. 12 is a schematic representation of an incubation system for the capillary tubes. 
     FIG. 13 is a schematic representation of an optical monitoring device for determining the position of fluid segments within the capillary tube. 
     FIG. 14 is an isometric, environmental view of a system for incubating a plurality of the capillary tubes in a controlled manner. 
     FIG. 15 is an enlarged, sectional view of a capillary tube employing a mixing compartment in a first alternate embodiment thereof. 
     FIG. 16 is a view similar to FIG. 15 of a second alternate embodiment of a capillary tube employing a mixing chamber. 
     FIG. 17 is a partial, sectional elevational view of an adapter for fluidly connecting a capillary tube with a motor driven precision lead screw mechanism. 
     FIGS. 18A,  18 B and  18 C show a schematic representation of a series of three steps for transferring a fluid droplet from a dispensing capillary to a receiving capillary in a method of the present invention. 
    
    
     DETAILED DESCRIPTION OF THE PREFERRED EMBODIMENTS 
     A high precision, small volume fluid processing system, in accordance with the principles of the invention, is generally indicated at reference numeral  20  in the Figures. With reference to FIGS. 1 and 2, the system includes a base  20 , an X-Y axis positioning mechanism  24 , a reagent dispensing station  26  and a sample station  28 . The base includes a pair of supports  30  which position a base platform  32  at a 45° angle with respect to a support surface  34 . The sample station  28  and reagent dispenser station  26  are positioned at 90° angles with respect to the platform  32  thus position the same at another 45° angle, with respect to the support surface  34 . As shown in more detail in FIGS. 3-5, the resulting spacial relationships positions the surface of a sample fluid  36  at approximately a 135° angle with respect to an open ended, hydrophilic capillary tube  38  for purposes which will be described in further detail below. 
     The X-Y positioning mechanism  24  includes a precision, screw actuated bed  40  for positioning an X axis stage  44  laterally with respect to the sample station  28  and reagent dispensing station  26 . An appropriated precision screw actuated bed  40  is available from Applied Precision, Inc., Mercer Island, Wash., under the model number AP 1000. The X axis stage has a Y motion frame  46  adapted for transverse motion with respect thereto so as to position the capillary tube  38  axially with respect to the sample station  28  and reagent dispensing station  26 . This is accomplished by mounting a stepper motor driven, computer controlled, precision linear actuator  48  on the X axis stage  44 , while an axially reciprocable portion thereof  49  is mechanically connected to the Y motion frame  46 . A computer or other suitable control device is connected to the actuator  48  by way of a control cable  50  in a manner to be described more fully below. An appropriate precision linear actuator  48  is available from Applied Precision, Inc., Mercer Island, Wash., under the brand name Nanomover™. 
     The Y motion frame  46  supports a sample fluid handling device generally indicated at reference numeral  52  which can precisely exspirate, and aspirate very small volumes of sample fluids, reagent fluids, etc. The device  52  includes a second, precision linear actuator  54 , the open ended hydrophilic capillary tube  38  previously described, and an adapter mechanism  56  for fluidly interconnecting the capillary  38  with the actuator  54  such that an internal spindle of the actuator acts as a high precision, pneumatic piston for controlling the position of a fluid segments in the capillary tube. A second control cable  58  operatively interconnects the precision stepper motor of the actuator  54  with a computer or other appropriate control device in a manner similar to that described with respect to the first precision linear actuator  48 . Both actuators contain relative or absolute position sensors  60 ,  62  which precisely determine the position of the stepper motor within the actuators, and thus the precise position of a fluid segment within the capillary tube  38 . 
     Preferably, the sample fluid handling device  52  has mounted thereon an optical position determining sensor  64  one portion of which has been removed for clarity in FIG.  1  and which is illustrated in further detail, schematically in FIG. 13 to provide a fluid segment position feedback loop for directly determining the position of a fluid segment within the capillary tube  38 . The sensor  64  is operatively interconnected with a computer or other appropriate control device through a third control cable  70 . 
     The system described above permits very small, precise volumes of fluid sample and fluid reagents to be precisely metered, mixed, and incubated for advantageous application to PCR, enzyme restriction digestion, ELISA, DNA sequencing, and other microbiological assays and processing techniques. The system can accurately handle fluid samples as small as 0.1 μl with an accuracy of ±0.01 μl. 
     This technique is further illustrated in FIGS. 3-8 in which the sample station  28  has been adapted to receive a conventional,  96  well microtiter plate  72  having a plurality of microwells  74  containing a volume (typically 5 ml or less) of a fluid sample  36 . The samples can be segments of DNA in a buffer solution from a single individual, or the like. As shown in FIG. 3, the capillary  38  is positioned laterally by the precision screw actuated bed  40  so as to be in alignment with one of the microwells  74  of the microtiter plate  72 . The capillary tube is then advanced in the direction of the arrow under the urging of the first precision linear actuator  48  until it is in contact with a lower level of the fluid  36  as shown in FIG.  4 . Sample fluid is then aspirated into the capillary tube  38  both by the natural capillary action of the tube, as well as by withdrawal of a spindle portion of the actuator functioning as a pneumatic piston as best seen with reference to FIG.  17 . 
     As shown in FIG. 17, the second, precision linear actuator  54  has a reciprocatable spindle portion  80  capable of precise axial movement in a manner well known to those of ordinary skill in the art. The actuator  54  has an extremely threaded end section  82  which has received thereon a cylindrical adapter  84  preferably manufactured from a nonreactive material such as stainless steel or heat treated aluminum. The adapter has a reduced diameter internal cavity  86  having a diameter of approximately 0.25 inch and an axial length of approximately 0.25 inch, and thus a volume of approximately 0.0128 cubic inches. The spindle portion  80  can protrude into this cavity so that the spindle and cavity form a precision pneumatic piston and cylinder. A forward, externally threaded end portion  88  of the adapter has a small diameter bore  90  therethrough for communication with an axial bore  92  of a threaded cap  94 . The cap is preferably manufactured from Delrin, or another suitably nonreactive thermoplastic material. A small diameter washer  96 , and “O” ring  98  are seated on an interface of the adaptor and cap so as to sealingly receive and seat the hydrophilic capillary tube  38  and to provide fluid communication between the capillary tube and bore  90 . A radially directed vent  100  is in fluid communication with the axial bore  92  to relieve pressure within the cavity  86 , bore  90  and bore  92  when the capillary tube is inserted into the cap  94 . 
     With reference again to FIGS. 3,  9 , and  8  is now apparent that upon retracting the spindle portion  80  in FIG. 17, a controlled volume of sample fluid  36  will be drawn into the capillary tube  38  so as to form a sample fluid segment  112  as shown in FIG. 5 when the capillary tube is withdrawn from the microtiter plate  72 . As further shown in FIG. 5, the position of the sample fluid segment  112  within the tube can be controlled by appropriate operation of the second precision linear actuator  54 . In this way, an air gap  114  may or may not be created so as to form a physical barrier between the segment  112  and any additional sample fluid segments which may be inspired into the capillary tube  38  for subsequent aliquoting purposes as shall be described further herein below. 
     A suitable capillary tube  38  has a length of approximately 55 millimeters, a maximum interior volume of 5 μl. The capillary has an inner diameter of 0.0134 inches. Tubes of this type are available from Drummond Scientific Company, Broomall, Pa., under the trademark MICROCAPS, part no. 1-00-0050-55. Capillary tubes of this type are naturally hydrophilic when clean. Thus, upon inserting the capillary tube  38  into the sample fluid  36  as shown in FIG. 4, a small amount of fluid sample will be drawn into the capillary without any movement of the spindle portion  80  of the second precision linear actuator  54 . In order to compensate for this “dead volume”, a correction factor must be applied to the command signal sent to the second actuator through the cable  58 . The dead volume V d  follows the formula: 
     
       
         
           V 
           i 
           =m V 
           d 
           +b.  
         
       
     
     The dead volume V d  may be calculated as:  V   d   =X   piston   k+V   d-min , where X piston  represent the position of spindle portion  80 , k represents a conversion factor from linear spindle position to displaced volume, and V d-min  represents the minimum dead volume inherent in the system. In order to aspirate a desired volume of sample fluid  36 , (V a ), the commanded aspiration is  V   a-command   =V   a   -V   l . 
     Using five μl capillaries and the Nanomover™ brand linear actuator  52 , adapter  56  and cap  94  described above, the quantities have the following values. 
     
       
         
           
               
               
               
             
               
                   
                   
               
               
                   
                 Quantity 
                 Value 
               
               
                   
                   
               
             
            
               
                   
                 m 
                 5.11 × 10 −3   
               
               
                   
                 b 
                 0.057 μl 
               
               
                   
                 k 
                 31.7 μl/mm 
               
               
                   
                 V d-min   
                 28 μl 
               
               
                   
                   
               
            
           
         
       
     
     This, for initial position of X piston =0, the dead volume, V d  is 28 μl and the initial take up due to capillary action is V l =0.2 μl. Therefore, for commanded aspiration of 0.1 μl of sample fluid  36 , this translated into a commanded aspiration of V a-command =−0.1 μl—a negative aspiration command. That is, in order to compensate for the capillary action of hydrophilic tube  38 , spindle portion  80  must move forward a distance equivalent to a volume of 0.1 μl. Larger desired volumes, V a  will of course require positive aspiration commands. 
     It is possible to avoid the dead volume problem described above by using a hydrophobic capillary tube. The clean glass capillary can be made hydrophobic by coating the interior thereof with silicone oxide or another material well known to those of ordinary skill in the art. A suitable product is available from Sigma Chemical Co., St. Louis, Mo., under the trademark SIGMACOTE, part no. SL-2. 
     Once the desired volume of sample has been aspirated into the capillary tube  38  as shown in FIGS. 3-5, the capillary tube  38  can be laterally moved into alignment with respect to the reagent dispensing station  26  to receive one or more appropriate reagents into the capillary tube for subsequent mixing with the fluid sample segment  112  of FIG.  5 . The preferred embodiment of the invention employs piezo electric reagent dispensers  120  as shown in greater detail in FIGS. 9 and 10. An appropriate reagent dispenser of this type is disclosed by Hayes, et al. in U.S. Pat. No. 4,877,745 the disclosure of which is incorporated herein by reference. It is sufficient for the purposes of this disclosure to explain that the piezo electric reagent dispenser has a reagent well  122  closed by a removable cap  124 . The well  122  contains an appropriate reagent  123  and is fluidly connected to a dispensing tube  126  having a dispensing orifice  128  at a free end thereof. The dispensing tube is concentrically surrounded by a piezo electric element  130  which when stimulated by an appropriate voltage generates a paristalic acoustic wave within the tube ejecting one or more droplets  132  of a reagent of known volume in a highly precise manner. 
     As shown in FIG. 6, the sample fluid segment  112  should be withdrawn a distance which corresponds to the volume of the droplets  132  and simultaneously with the expulsion thereof from the dispensing tube  126  so that the droplets do not encounter back pressure within the capillary tube. The result will be a second fluid segment  134  consisting exclusively of reagent fluid adjacent to the sample fluid segment  112 . If an air gap  114  as shown in FIG. 5 is desired the series of segments within the capillary tube  38  shown in FIG. 7 will be as follows: reagent fluid segment  134 , air gap segment  114 , and the sample fluid segment  112 . In either event, by oscillating the segments within the capillary tube  38  in opposite directions with differential velocities V 1 , V 2  the fluids will mix within the capillary tube. To ensure adequate mixing, one of the velocities should be at least three times the other velocity, and the mixing should occur over 100 cycles at a frequency of three cycles per second. In a five μl capillary when 3 μl of fluids have been received, axial oscillation of the fluids, of ±1.5 μl can occur without inadvertently aspirating the fluids from the free end of the capillary tube  38 . 
     In order to accurately position the fluid segments with the capillary tube  38 , an optical position measuring device  64  as shown in FIG. 1 is preferably provided although all of the sequencing steps described above can be and have performed in an open loop mode. The device  64  is further described schematically in FIG. 13 wherein a printed circuit board  140  supports a charge couple device  142  thereon for sensing the position of three exemplary fluid segments  144 ,  145 , and  146  in the capillary tube  38 . An array of light emitting diodes  148  illuminates the fluid segments within the capillary tube  38 . Poto diodes detects the presence of light transmitted through air gaps  151 ,  152 , and  153  and outputs this information to a charge coupled device which then transfers this information serially to a digital computer or other appropriate control device. As shown in the schematic representation at reference numeral  154 , a graph of signal amplitude on the vertical axis versus segment position with respect to the free end  136  of the capillary tube  38  indicates not only the presence or absence of the fluid segments but their respective leading and trailing edges as well. Using standard interpolation techniques, resolutions of up to 50 nanometers of absolute position have been achieved by using a charge couple device array  42  manufactured by Sony under the model no. IFX503A. A development kit is included with this part which will enable those of ordinary skill in the art to execute the design shown in FIG. 13 without undue experimentation. 
     With reference to FIG. 11, it is seen that an appropriately programmed personal computer  156  employing conventional analog to digital controller modules (not shown) can be operatively interconnected with the precision screw actuated bed  40 , first precision linear actuator  48 , second precision linear actuator  54 , piezo electric reagent dispensers  120  and optical position determining sensor  64  so as to comprise a fully automated precision small volume fluid processing apparatus. In addition, a capillary dispensing station  160  having a V shaped capillary trough  162  can be used to position individual blank capillary tubes in a receptacle  164 . As will be apparent from examination of FIG. 17, it is possible for the second linear actuator  48  to advance the sample fluid handling device linearly towards the receptacle  164  (which is shown in elevational view for clarity) so as to insert a blank capillary tube  38  into the cap portion  94 . The computer can then command the precision screw actuated bed  40  to position the second actuator  54  in alignment with the sample station  28  and receive a predetermined volume of sample as previously described. The optical position determining sensor  64  can then send position information to the computer  156  to confirm that the correct volume of sample fluid has been aspirated and positioned appropriately within the capillary tube  38 . The computer can then command the bed  40 , and actuator  48  to move the capillary tube  38  to be moved into appropriate positioning  38 ″ with respect to the reagent dispensing station  26  to receive one or more precisely predetermined volumes of reagent. Once the reagent(s) and fluid sample have been appropriately mixed within the capillary tube  38 , the computer can command the capillary tube  38  to be moved to a new position  38 ′″ for further processing. 
     As shown in FIG. 12, further processing can include incubating the capillary tube  38  in an appropriate temperature controlled environment. This result can be achieved by providing a thin, substantially transparent, highly resistive coating  170  on the exterior of the capillary and connecting electrodes  172 ,  174  to an appropriate voltage source  178  which is in operative communication with the computer  156 . Appropriate coating is indium tin oxide applied by the conventional vapor deposition techniques. It is known that the resistivity of a thin film of indium tin oxide changes with temperature, thus a continuous resistive measurement to be made by an electronically controlled Ohm meter  178  also operatively connected with the computer  156  can be used as a measurement of the tube temperature. A cooling fan  180  is preferably positioned advantageously with respect to the capillary tube  38  provide a constant air flow over the tube and a rapid cool down cycle when the voltage is no longer applied across the electrodes  172 ,  174 . A conventional temperature sensor such as Thermistor type temperature sensitive resister  182  can also be preferably operatively interconnected with the computer to measure the temperature of the air flow exiting the fan  180 . 
     FIG. 14 illustrates a preferred embodiment for implementing the incubation technique shown schematically in FIG.  12 . Before the incubation step shown in FIG. 12 can proceed, both the free end  136  and an open mounting end  137  of the tube  38  should be heat sealed in the conventional manner. Once this has been done, a plurality of the sealed, capillary tubes  36  can be incubated by placing them in a holder, generally indicated at reference numeral  190  in FIG.  14 . The holder includes opposed end caps  192 ,  193  having the electrodes  172 ,  174  therein and in communion with appropriately sized apertures  194  for receiving the capillary tubes  36 . The exterior ends of the capillary tubes are preferably plated with chrome or another conductive material to make good electrical contact with a peripheral copper strip  196 ,  197  connected to the electrodes  172 ,  174 . A space block  200  provides a mechanical support for the end caps to prevent the tubes from being crushed. In this preferred embodiment, the cooling fan  180  and Thermistor™ are contained in a plenum or duct  210  which directs the cooling air over and through the capillary tubes  36  for the purposes previously described. 
     As described above, the capillary tube  38  is preferably a hydrophilic tube. Moreover, as previously stated, the tube may be made hydrophobic by the introduction of appropriate materials. FIG. 15 shows an alternate preferred embodiment of a hydrophobic capillary tube  220  having an enlarged diameter mixing chamber  222 . In this embodiment, the enlarged chamber in combination with an interior hydrophobic surface  224  increases turbulence in the tube during mixing and thus improves the mixing process. One technique for manufacturing such a tube is shown in FIG. 16 in another alternate embodiment in which two conventional sections of hydrophilic capillary tube  38  are joined in a spaced apart relationship by a larger diameter section  230  of a hydrophobic capillary tube section. 
     In the case of either embodiment shown in FIG. 15 or  16 , these capillary tubes can be used in the same manner as shown with respect to the preferred, hydrophilic tube. Finally, it should be noted that a capillary tube which is neither hydrophilic or hydrophobic can further be used. 
     FIG. 18 illustrates an aliquoting method using the system shown in FIG. 10 wherein the reagent dispensing station  26  is provided with one or more hydrophilic receiving capillary  300  in place of the reagent dispensers  120 . In this method as shown in FIG.  18 ( a ) one or more segments  112  of sample fluid have been inspired into the capillary tube  38  and are separated by an air gap  114 . By advancing the spindle portion  80  a desired amount a droplet  310  of known volume can be formed at the free end  136  of the capillary tube  38 . As an example of the preferred embodiment given above, each step taken under computer control by the stepper motor of the actuator  54  corresponds to an increase in the size of the droplet  310  of 1.583 nanoliters. When an appropriately sized droplet is formed, the capillary tube  38  is advanced towards the receiving tube  300  by the first linear actuator  48  as shown in FIGS. 1 and 2. As shown in FIG.  18 ( b ) as soon as the droplet contacts the free end  320  of the receiving capillary  300  the droplet  310  of known volume will be drawn into the receiving capillary  300  by capillary action. The steps shown in FIG.  18 ( a ) and  18 ( b ) can be repeated until a sample fluid segment  330  having a volume equal to or less than the fluid segment  112  has been transferred. Droplets as small as 50 nanoliter increments have been transferred using this technique with the capillaries described. Once all of the sample fluid segment  112  has been transferred, the air gap  114  prevents the next sample fluid segment is capillary  38  from being inadvertently transferred. The capillary  38  may then be laterally moved to a second receiving capillary (not shown) and the process may be repeated. In this manner, the capillary tube  38  can be used to aliquot DNA samples into very small volumes, and distribute those volumes into separate receiving capillary tubes  300  which may then be individually processed with the appropriate as described herein above. 
     EXAMPLE I 
     The apparatus described above has been used to perform a restriction enzyme digest as described below. 
     In restriction enzyme digest (RED), a DNA sample is mixed with a “restriction enzyme” in a buffered solution and incubated. Each restriction enzyme will cleave that DNA at each site containing the sequence of base pairs specific to that enzyme. Restriction enzymes are sold commercially. An enzyme and DNA segment were chosen known to produce multiple cuts. 
     The DNA fragment sizes are determined by gel electrophoresis after the incubation. In this procedure, the (digested) DNA is placed on a gel (typically agarose) which is immersed in an electric field. The DNA will migrate through the gel at a rate determined by it&#39;s size: big pieces are slower. 
     First, an enzyme solution was prepared containing “Hind III” restriction enzyme, a buffer sold with the enzyme which is specific to that enzyme, sterile water, and an extra protein known as “BSA”, according to the following recipe: 
     15 parts sterile, double distilled water 
     2 parts buffer 
     1 part BSA (2.5 mg/μl) 
     1 part enzyme (Hind III) 
     The BSA is necessary to keep the enzyme in the solution. Enzymes, like all proteins, tend to adhere to surfaces such as the inside of capillary tubes, test tubes, the pipette tips, and the reagent dispensers  120 . The BSA is used as a sacrificial protein to occupy the adhesion sites and help keep the enzyme in solution. Gelatin and BSA are both commonly used for this purpose in micro-biology/genetics procedures. 
     The DNA chosen for this experiment was commercially prepared “Lambda” DNA, GibcoBRL part #25250-010, concentration 0.52 ug/μl. The enzyme chosen was “Hind III”, GibcoBRL part #15207-012, concentration 10 U/μl, which comes with it&#39;s own specific buffer (GibcoBRL “React 2” buffer). 
     Starting with a clean, sterile container, 1 part DNA was added with 19 parts of the enzyme solution listed above. The resulting mixture was agitated, then incubated at 37 C for two hours. The volumes were reduced using the method of the invention by a factor of 10, to 0.1 μl of DNA and 1.9 μl of enzyme. 
     A large batch of the enzyme mix, 200 or 300 μl worth was mixed up with the excess and used to prime the reagent dispenser  120 . The capillaries were then flame sealed with a butane torch prior to incubation. 
     Next the samples are removed from their capillary and placed on a 2%, agarose gel of the “high melt” variety—commonly known as “2% HMP agarose”. The gel is exposed to an electric field, around 100-120 volts, for 15-20 minutes. 
     After electrophoreses, the gel is soaked in an Ethidium Bromide solution to stain the DNA, and examined under a UV light. DNA will show up as florescent bands. 
     All but the two smallest DNA bands could be identified. GibcoBRL note in their documentation these two bands are hard to distinguish because they are so much smaller than the largest fragment sizes, and this same caveat applies to their reference. 
     The bands from the samples matched the bands in the GibcoBRL reference, with the two smallest bands visible only faintly, even in the reference. The second smallest band was made more visible in the reference which was intentionally overloaded a gel lane with excess DNA. 
     Other embodiments and variations of the invention are contemplated. Those or ordinary skill in the art will indeed conceive of other embodiments and variations of the invention upon review of this disclosure which are not herein described but are within the spirit of the invention. For example, it is contemplated that the receiving capillary  300  can be hydrophobic and can have an open mounting end (not shown) fluidly connected to a third precision linear actuator, similar to linear actuators  48 ,  54 , by way of an additional adaptor  84  and cap  94 . The third linear actuator can be coordinated with the first actuator which drives the dispensing capillary  38  in FIG.  18 ( a ). In this way, the droplet  310  can be aspirated into the receiving capillary under positive control rather than by capillary action. Therefore, the invention is not to be limited by the above disclosure, but is to be determined in scope by the claims which follow.