CA1286389C - Automatic pipetting apparatus - Google Patents

Abstract

TITLE: AUTOMATIC PIPETTING APPARATUS

ABSTRACT
An automatic pipetting apparatus is disclosed having a base on which is mounted a vertical frame supporting a row of positive displacement pipettes which are driven in the up or down direction by an electromechanical mechanism under microprocessor controls. The apparatus includes a bass, a track and a carriage longitudinally movable beneath the row of pipettes. The carriage includes an independent translating and position signal generation mechanism. The carriage carries a tray which includes sample chambers, a space to receive a microporous electrophoresis support medium, such as a cellulose acetate strip, a wash well, rinse well and a space to receive blotting paper. The pipettes include a barrel and a plunger capable of aspirating and dispensing from .5 to 5 µl of liquid. The barrels move up and down with respect to the base by means of another independent translating and signal generating mechanism. The barrels may be easily replaced from the mechanism if they become damaged or worn from many operating cycles. The apparatus under microprocessor program control, washes, rinses, blots the barrels before and after each application or engagement of the barrels with a liquid or contaminant.

Description

3~9 AUTOMATIC PIPETTING APPARATUS

BACKGROUND OF THE INVENTION
Field of the Invention This invention is related in general to the field of applylng fluid samples to analysis strips. In particular, this invention relates to an automatic pipetting apparatus for applying multiple fluid samples to a microporous ~0 support medium such as a cellulose acetate or agarose strip ~hich may be used in the field of zone electrophoresis and with other separation techniques including the field of thin layer chromatography. Zone electrophoresis is the science of moving charged particles ~n an electric field through a solid or semi-solid medium.
The technique is most commonly used in medical research and medical laboratories for analyzing various blood proteins.
Description of the Prior Art ~n the electrophoresis technique, a blood or other fluid sample is applied to a support medium ~hich is then subjected to an electric field so as to separate the components of the sample. The support media used in the electrophoresis process includes cellulose acetate, agar, agarose and acrylamide gels. In laboratory work it is desirable that a plurality of samples be applied to the 3~31'3 support medium such that each of the samples may be subjected to the electric field at the same time.
Tho samples may be applied to the support medium one at a time in serial fashion ~ith a hand pipetter, but the hand pipetter must be rinsed ~ith a cleansing agent and blotted before a ne~ sample is aspirated and then applied to the strip.
Applicators have been designed to apply fluid samples simultaneously or in "parallel" to the strips. Such applicators are described at page 61 of the General Products Catalog for 1984-19B5 of Helena Laboratories ~ith offices in Beaumont, Texas. Such applicators may apply eight, t~elve or more samples to a microporous support medium and have the advantage of making the electrophoresis technique easier and more reproducible.
The applicators kno~n prior to this invention ho~ever have been essentially non-automatic applicators and required cleaning of the applicator tips after each application to the support medium.
Automatic dispensing systems are known in the prior art. For example, a system sold under the trademark "~Tell Washers" of BioTech Instruments, Inc. of Burlington, Vermont provides an alignment mechanism by ~hich a ro~ of eight or t~elve barrels may be positioned above one of a plurality of ro~s of ~ashing vials or wells. Automation in the system provides selection of dispensing fill volumes, soaking times and number of wash cycles.

12~36389 None of the prior art ho~ever has provided an apparatus ~or automatically filling a plurality of pipetters from a respective plurality of fluid chambers and then precisely applying such fluid samples from each pipette to a support medium. Another disadvantage of the prior art systems is that there has been no means for automatically ~ashing and cleaning the barrels during each cycle time so as to prevent contaminatio~ of each of the barrels during application of a ne~ plurality of fluid samples to a ne~ support medium.
Another disadvantage of the prior art is that there has been no means for precisely automatically applying a very small amount - of the order of one micro liter of sample liquid - to a support medium.
Another disadvantage of the prior art is that there has been no means for precisely automatically diluting a very small amount - of the order of one micro liter - of sample fluid ~ith a diluting liquid, and precisely applying a very small amount of the diluted sample to a support medium.
Accordingly the invention seeks to provide an automatic pipetter apparatus for aspirating from a plurality of sample chamber wells into a corresponding plurality of pipetters and then applying such samples precisely to~ a microporous support medium to be used in electrophoresis or thin layer chromatography.

,' ', ., ' . , - :, , ~Z~3ti3~39 Further the invention seeks to provide an automatic pipetting apparatus ~hich not only aspirates and dispenses sample fluids onto a support medium strip such as cellulose acetate or agarose, but also flushes, cleanses, rinses and blots the tips of the barrels ~ith an appropriate cleaning fluid before and after each application of the sample fluid to the support medium.

Still further the inve~tion seeks to provide an automatic pipetting apparatus by ~hich p~sitive displacement pipette barrels and plungers are controlled to precisely apply a very small sample of fluid to a support medium.

Further still the invention seeks to provide automatic plpetting apparatus for precisely automatically diluting a very small amount of sample fluid with a diluting liquid and precisely applying a very small amount of the dlluted sample to the support medium.

SUMMARY OF THE INVENTION
-Accordingly, in one embodiment of the invention there is provide,d an automatic pipetting apparatus ~hich generally includes a base and a sample plate disposed on the base and a pipette frame including a vertical support for supporting the frame from the base above the sample plate. The sample plate includes a ro~ of individual liquid sample chambers and a lateral application space longitudinally 3,~3 separated from the li~uid chamber ro~. The lateral application space is adapted to receiv~ a microporous support medium. A mounting plate is carried by the pipette frame.
The apparatus includes translation means for effecting relative longitudinal movement of the pipette frame and sample plate, and vertical translation means for effecting rela ive vertical movement of the mounting plate and the sample plate.
A plurality of microsyringe barrels are removably ~ecured to the mounting plate. The barrels are spaced corresponding to the spacing of the liquid chambers of the sample plate. A plurality of micro-plungers are provided, one each movably disposed in one of the barrels. A
plunger translation means is provided for moving the plunger vertically within the barrels.
Signalling means are provided for generating longitudinal signals representative of the relative longitudinal orientation of the pipette frame with respect to the sample plate, for generating mounting plate signals representative of the vertical orientation of the mounting plate relative to the sample plate and for generating plunger signals representative of the orientation of the plungers relative to the barrels.
A programmed digital computer is provided responsive to the longitudinal signals, the mounting plate signals and to the plunger signals for generating a sequence of , 128G3~39 control signals to the longitudinal translation means, to the vertical translation means and to the plunger translation means to aspirate a predetermined amount ~f liquid ~rom the sample chambers into the respective pipette barrels, and to apply the liquid samples in the barrels onto corresponding spaces or "spots" on the microporous support medium when placed on the lateral application space of the sample plate.

~nother embodiment of the invention includes a base having a track disposed longitudinally on it. A carriage i8 longitudinally movably disposed on the track and carries a sample plate ~hich is removably disposed on the carriage. The carriage plate includes a lateral ro~ of individual liquid sample chambers and a lateral application space ~hich is longitudinally separated from the liqu~d chamber ro~. The 1ateral application space is adapted to receive a microporous support medium such as a cellulose acetate or agarose strip used in electrophoresis or thin layer chromatography.
A pipette assembly is mounted vertically on the base above the carriage and the sample plate. The pipette assembly includes vertical mounting posts separated laterally from each other and secured to the base. A
mounting plate assembly is slidably guided by the posts and is disposed laterally ~ith respect to the sample plate.

.. . . . .

3~3 The mounting plate assembly includes a mounting plate having slidable guides disposed about the posts. A
pipette bar is fixed to the mounting plate. A plurality of microsyringe barrels are provided in a ro~ on the pipette bar ~ith their heads secured thereto~ The microsyringe barrels are spaced corresponding to the spacing of the liquid chambers on the plate. The barrels are hollo~, each barrel having a lower tip.
A plunger bar is vertically movably disposed above the tip bar and has a plurality of micro-plungers secured thereto. Each of the micro-plungers are movably disposed ~ithin a corresponding barrel of the microsyringes. A
plunger actuator plate carried by the mounting plate is vertically movable ~ith respect to the mounting plate.
The actuator plate is removably secured to the plunger bar.
Translation and signalling means are provided for moving the carriage longitudinally for~ard and back~ard beneath the mounting plate assembly and generating carriage position ~ignals indicative of the carriage position. A translation and signalling means is provided for moving the mounting plate assembly up and do~n ~ith respect to the base and generating mounting plate position signals indicative of the mounting plate position. A
translation and signalling means for moving the plunger bar up and do~n ~ith respect to the mounting plate and lZ~j3fi~

generating plunger bar position signals indicative of the plunger bar position is provided.

A programmed microcomputer is provided responsive to the carriage position slgnals, to the mounting plate position signals, and to the plunger bar position signals for generating a sequence of control signals to the translation means for moving the carriage, the translation means for moving the mounting plate, and the translation means for moving the plunger bar so as to aspirate a first predetermined amount of liquid from the sample chambers into the respective pipette barrels and then to apply the liquid in each of the pipette barrels onto corresponding spaces of the support medium when placed on the lateral application space of the sample plate.

Another aspect of the invention provides a sample plate adapted for use with automatic pipetting apparatus comprising a lateral row of individual liquid sample chambers and a lateral application space longitudinally separated from the liquid chamber row. The lateral application space is adapted to receive a microporous support medium and includes a raised portion and a lower portion, the row of individual liquid sample chambers being disposed on the raised portion, and the lateral application space disposed on the lower portion and further comprising a wash well and a waste well longitudinally spaced from each other and from the sample chambers.

A further aspect of the invention comprehends a sample plate adapted for use with automatic pipetting apparatus . ~ .
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~Z~36389 comprising a lateral row of individual liquid sample chambers and a lateral application space longitudinally separated from the liquid chamber row, the lateral application space being adapted to receive a microporous support medium, and further comprising a wash well and a waste well longitudinally spaced from each other and from sample chambers. There is further a longitudinal blotting space for applying a lateral blotting paper strip, the blotting space longitudinally separated from the sample chamber row, the wash well, the waste well and the lateral application space.

Still another aspect of the invention pertains to apparatus adapted for use with automatic pipetting of liquid samples to be subjected to electrophoresis comprising a sample plate including a lateral row of individual liquid sample chambers and a lateral application space longitudinally separated from the liquid chamber row, the lateral application space having disposed thereon a removable microporous support medium adapted for electrophoretic analysis after a liquid sample has been deposited thereon. The sample plate includes an upper portion and a lower portion, the row of individual liquid sample chambers being disposed on the raised portion and the lateral application space being disposed on the lower portion.

BRIEF DESCRIPTION OF THE DRAWINGS

The aspects, advantages and features of the invention will become more apparent by reference to the drawings which are appended hereto and wherein like numerals indicate like parts and wherein an illustrative embodiment of the invention is shown, of which:

3L2~i~;389 Figure 1 shows a perspective vie~ of one embodiment of the automatic pipetting apparatus of the invention with a sample plate secured thereto and its cover attached;
Figure 2 is a schematic illustration of the invention sho~ing the functional relationship bet~een the sample plate with its sample chambers, wash well, ~ast~ well and a longitudinal application space with a microporous support medium secured thereto beneath a pipette assembly in which individual pipette barrels are moved as a unit up and do~n and a plunger bar secured to plungers which are moved up and down ~ith respect to the barrels;
Figure 2A schematically shows another embodiment of the invention where the mounting plate includes two stationary units and the pipette frame is longitudinally movable with respect to the plate;
Figure 2B i8 a perspective view of another embodiment of the sample plate further including a ro~ of dilution wells;
Figure 3 i8 a side view partially cut a~ay and in section ~ith the cover removed and taken from the view along lines 3-3 of Figure l;
Figure 4 is a cross-section of the carriage and the sample plate;
Figure 5 is a downward looking view along lines 5-5 of Figure 1 and shows the cross-section of the base at level 5-5 and the track, carriage ~nd sample plates beneath the pipetting assembly;

3~9 Figure 6 is a cross-sectional vie~ taken along lines 6-6 of Figure 5 and sho~s the bas~, the carriage and sample plate according to the invention and further sho~s the translatio~ and guiding means by ~hich the track is moved for~ard and back~ard ~ith respect to th~ pipetting assembly;
Figure 7 is a for~ard looking vi~ taken along lines 7-7 of Figure 3 and sho~s partially broken a~ay, partially cross-sectional parts of the pipette assembly slidably mounted on posts secured to the base and including a mounting plate ~hich moves up and do~n ~ith raspect to the base and carrying a ro~ of barrels of individual pipettes and an actuator plate movable vertically ~ith resp~ct to the mounting plate for moving a plunger bar vertically for moving individual plungers within the barrels of each of the pipettes;
Figure 8 sho~s a top vie~ of the pipette assembly showing in dashed lines the mounting platP of the pipette assembly;
Figure 9 shows a cross-sectional vie~ taken along lines 9-9 of Figure 7 and illustrates the relationship bet~een the mounting plate, the tip bar and the plunger bar and the means by ~hich the mounting plate is moved up and do~n ~ith respect to the base;
Figure 10 is a similar vie~ to that of Figure 7 but sho~s the mounting plate having been translated to a lo~er position but ~ith the plunger bar remaining in an up~ard 1~13G3~9 position ~hereby the tips of the pipetters are in a do~n~ard position, but the plungers are extended up~ardly from each of the barrels of the pipettes Figure 11 is a top cross-sectional vie~ taken along lines 11-11 of Figure 10 and illustrates the motor and rack and pinion system by ~hich the mounting plate is moved up and do~n ~ith respect to the base;
Figure 12 is a cross-sectional vie~ taken along lines 12-12 of Figure 10 and illustrates the relationship of the mounting bar after it has been moved do~n~ardly by the rack and pinion system by operation of the motor turning ~ith re~pect to the frame mounted rack;as shown with Fig. 9;
Figure 13 is a vie~ similar to that of Figures 7 and 10 but illustrates the plunger actuator plate and plunger bar moved do~nwardly with respect to the mounting plate thereby forcing the plungers associated ~ith each of the pipetters into their barrels and forcing any fluid previously aspirated into the barrels out the tips of the barrels;
Figure 14 shows a cross-sectional vie~ taken along lines 14-14 of Figure 13 and illustrates the rack and pinion system by ~hich the actuator plate and the plunger bar secured thereto is moved up and do~n ~ith respect to the mounting plate;
Figure lS is a cross-sectional vie~ taken along lines 15-15 of Figure 13 and illustrates the movement of the actuator plate do~n~ardly ~ith respect to the mounting plate by operation o~ the rack ana pinion system controlling relative movement bet~een the actuator plate and the mounting plate; as shown with Fig. 9;
Figure 16 is a schematic illustration of the microcomputer integrated circuit device receiving signals from position detector circuits associated ~ith the carriage, the mounting plate and actuator plate and applying translation signals to motors for positioning the carriage, the mounting plate and the plunger actuator plate; and Figure 17 is a functional flow chart illustrating the stored program in the microcomputer device for automatically washing, blotting, sampling and applying samples to the support medium strip.

DESCRIPTION OF THE INVENTION
. _ Figure 1 illustrates in a perspective vie~ the automatic pipetting apparatus 10 according to the invention. The apparatus includes a base 12 on ~hich a sample plate 16 is movably supported. The sample plate includes a space 21 for securing a microporous support medium 20 such as a cellulose acetate or agarose strip used in the field of zone electrophoresis or other separation techniques including the field of thin layer chromatography. A cover 14 is provided behind ~hich a pipette head 18 is sho~n.

3~

Figure 2 is a schematic illustration of th~ essential mechanical elements of one embodiment of the invention ~ith the base and the cover removed. None of the mounting apparatus is sho~n in Figure 2 so as to simplify the explanation of the relationship of the sample plate 16 to the pipette assembly 70. The translational means are sho~n in a functional way rather than in actual mechanical detail ~hich ~ill be sho~n in detail in the figures and discussion below.
The sample plate 16 as sho~n includes a row of sample chambers 24 as ~ell as a wash well 26, rinse or "waste~
well 28 and a longitudinal space 21 on which a microporous support medium 20 is removably secured. The sample chambers 24, the rinse ~ell 28 and the wash ~ell 26 are provided on raised portions 22 of the sample plate. If desired, a plastic cup may be provided in each of sample chambers 24. A blotter space 32 between the rinse ~ell 28 and the sample chambers 24 is provided at substantially the same vertical level 49 as the sample application space 21 on ~hlch the support medium 20 is secured.
As illustrated in Figure 2, the various regions of the sample plate are longitudinally distinct, yet the lateral spacings bet~een the sample chambers 24 corresponds to the application spots 19 on the support medium 20 which is indicative of the fact that the barrels 92 of the pipette assembly 70 are arranged in a row corresponding to the sample chambers 24. Liquid from : .
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~Z~3~3~

those chambers is aspirated by the automatic pipetting apparatus and is applied in a similar ro~ on the spots 19 of the microporous support medium 20.
It is advantageous to provide the sample chambers 24, ~aste ~ell 28 and wash ~ell 26 in raised portions 22 of the sample plate 16 so that the mounting plate 80 of the pipette assembly 70 need only go down to a common do~n~ard position during all wash, ~aste, blot, ~ampling and application operations. Ho~ever, it ~ould be obvious to one of ordinary skill in the art that other arrangements could be provided e~pecially ~here different levels of the mounting plate could be prov~ded in the translation and signalling apparatus for controlling the mounting plate 80. A detailed discussion of such translation and signalling apparatus for controlling the mounting plate 80 i8 discussed belo~.
The schematic illustration of Figure 2 sho~s that the sample plate 16 i~ tran~lated in for~ard and rearward directions beneath the pipette assembly 70 by virtue of the motor 40 turning a pinion 38 having its gears in engagement with those of rack 36. As the shaft of the motor 40 turns, the sample plate 16 carried by the rack 36 moves back and forth beneath the pipette assembly 70.
Turning no~ to the pipette assembly 70 sho~n in Figure 2, a mounting plate 80 is translated up~ardly and do~n~ardly by means of mounting plate motor 116 having its pinion 120 engaging a rack 122. Thus, the entire mounting '12~3~3F~9 plate 80, and the microsyringe barrels 92 attached to the barrel bar 88 which is secured to the mounting plate 80, moves up and down in accordance with the turning of the mounting plate motor 116. Similarly, the plungers 94 which are attached to plunger bar 90 and actuator plate 84 are moved up and do~n ~ith respect to mounting plate 80 by operation of the turning of actuating plate motor 126 and its pinion 130 engaging actuator rack 132. For purposes of illustration, the actuator plate motor 126, its pinion 130 and the actuator plate rack 132 are sho~n on the forward side of mounting plate 80, but the actual apparatus illustrated in the subsequent figures is to the rearward side of the mounting plate 80 through slots in it.
Figure 2 therefore sho~s all of the essential elements as far as the translation of the sample plate 16 backward and forward beneath the microsyringe barrels 92 and illustrates the upward and downward translation means of the mounting plate 80 and the microsyringe barrels 92, and the upward and downward motion of the plungers 94 and the plunger bar 90 and actuator plate 84 with respect to the mounting plate 80.
Figure 2A schematically illustrates an alternative embodiment of the invention where the sample plate remains immovable with respect to the base 12' ~ith the pipette assembly 70' being mounted on rollers 300 for longitudinal translation. Figure 2A illustrates that the sample plate i~Z~3?39 may include two units, an application plate unit 16A and a fluid plate unit 16B. The sample plate unit 16A is adapted to removably secure a support medium 20', while the fluid plate unit 1 6B includes a ro~ of sample chambers 24', a ~aste well 28', a ~ash well 26l and a blotting space 32'. The operation of the alternat~ve embodiment is similar to that of the embodiment of Figure 2 except ~hat translation and signalling means are provided for longitudinally translating pipette assembly 70' ~ith respect to the sample plate(s) 16A, 16B. Details of such translation and signalling means will be apparent to one of ordinary skill in this art by virtue of the detailed description of analagous translation and signalling means described below.
Figure 2B illustrates an alternative sample plate 16' ~hich may include an additional ro~ of dilution wells 25 in addition to the row of sample chambers 24, waste ~ell 28, ~ash ~ell 26 and blotting space 32. Explanation of the automatic diluting of sample fluid will be described below with reference to Figure 2B.
Figure 3 is a side vie~ taken along lines 3-3 of Figure 1 ~ith a portion of track 34 cut a~ay to show its construction. The tracks 34 are supported by track supports 48 which may also be seen in Figure 5. The pipette assembly 70 is vertically supported from base : mounting block 78 which is secured to the sides of the . base 12 and is also further illustrated in Figure 5. The lZt~ 9 pipette assembly 70 includes a back plate 86 and a front plate 73. One of the plurality of barrels 92 of the pipette assembly is shown in an up~ard position.
A carriage 46 is slidably movably disposed on track 34 as more clearly seen in Figure 6. Racks 36 are secured to carriage 46 and are movable with respect to the base 12 by means of the carriage motor 40 having its pinion 38 in engage~ent with rack gear 36.
Notches are provided along the left edge of the carriage 46. These notches cooperate ~ith a trip switch to provide signals indicative of the longitudinal position of the carriage. The wash notch 50, rinse notch 52, blot notch 54, sample chamber notch 56, dilution chamber notch 57 (where the alternative sample plate 16' of Figure 2B is used) and application notch 58 are illustrated in Figure 3.
Figure 4 illustrates a vertical cross-section through the carriage 46 and the plate 16 and shows the actual ~ash well 26, rinse well 28 and one of the sample cham~ers 24 on raised portion 22 of the plate 16. A blotter space 32 and a lateral application space 21 are illustrated on plate 16. Blotter paper 30 is shown in blotter space 32 while a microporous support medium 20 such as cellulose acetate or agarose is secured in lateral application space 21.
Figure 5 is a do~nward view taken along lines 5-5 of Figure 1. Carriage 46 is shown supported by tracks 34 and 38~
~20~

movable in the rearward and for~ard directions by means of motor 40, pinion 38 and rack 36 as also illustrated in Figure 5. The sample plate 16 is disposed in a valley or IJ-shaped cross-sectional structure 47 of carriage 46.
Position signalling notches on the sid~s of the carriage 46 cooperate with trip s~itch 60 and trip switch 62 fixed to tracks 34. Spring loaded rollers 61 and 63, respectively are forced against the longitudinal edges of carriage 46 and into the notches as the carriage 46 moves pa~t them. For example, the notches on the left hand side 46 include the wash notch 50, the ~aste notch 52, the hlot notch 54, the sample chamber notch 56 and an application notch 58. The notches correspond to the longitudinal position of the wash ~ell 26, waste well 28, blotter paper 30, sample chambers 24 and application spots 19 when those wells, chambers and blotting and application spots are directly beneath the pipette barrels 92.
When the carriage 46 moves rearwardly ~here the wash ~ell 26 is directly beneath the barrels 92, the roller 61 moves into the ~ash notch 50 thereby tripping the trip switch 60 for signalling the microprocessor associated with electronic module 200 (Figures 3 and 16) that the wash well is beneath the barrels 92. The trip switch 60 is likewise tripped when the roller 61 enters notches 52, 54, 56 and 58 to signal the position of the respective other wells, chambers and spots beneath the barrels 92.

l Z~3B9 On the right hand side of the carriage 46 is notch 59 in ~hich the roller 63 is sho~n. A sample plate trip s~itch 62 is thereby tripped to indicate that the carriage is at its maximum forward position. Trip s~itch 65 is mounted on the rear edge of carriage 46. Switch 65 closes ~hen the rear edge of the plate 16 is in position and engages it thereby generating a signal that the plate 16 is properly in position on carriage 46. The stops 64 provide means for accurately longitudinally positioning application spots 19 beneath barrels 92 ~hen carriage 46 is`in the maximum rear~ard position.
Also shown in Figure 5 is a start button 214 by which the programmed microprocessor is signalled to start the automatic sequence of events for the automatic pipetting apparatus ~hich ~ill be explained in detail belo~. Lamp 212 provides a visual indication to the user of the automatic pipetting apparatus 10 that the po~er is turned on.
Figure 6 illustrates in a cross-sectional vie~ taken along lines 6-6 of Figure 5 the means by ~hich the carriage is translated ~ith respect to tracks 34. The tracks 34 are supported upon base 12 by means of supports 48. The carriage 46 includes slots 149 in its sides on ~hich it slides on tracks 34.
Plate 16 as indicated above is disposed ~ithin a notch or valley 47 of the carriage 46. The carriage translation motor 40 is fixed to the base 12 and includes ~2 ~ 3 a shaft 42 ~hich is supported by means of shaft supports 44. Pinions 38 secured to shaft 42 have their gears in engagement ~ith racks 36 ~hich are attached to the carriage 46. As the motor 40 is turned in one direction or the other under control of the microprocessor in the electronic module 200 (Figure 16), the carriage 46 moves in the for~ard or rear~ard directions.
Figure 7 illu~trates the pipette assembly 70 looking rear~ardly along lines 7-7 of Figure 3. The mounting blocks 76 are sho~n secured to the base mounting blocks 78 by means of scre~s 79. The mounting blocks 76 carry vertical mounting posts 72 as illustrated in Figures 7, 10 and 13 and in the top view6 of Figures 8, 11 and 14. The front plates 73 and back plate 86 are secured by means of scre~s to mounting blocks 76.
A mounting plate 80 is vertically slidably supported about the vert$cal mounting po~ts 72. Retainer bearings 74 provide sliding engagement bet~een the posts 72 and vertical bearing blocks 75. The mounting plate 80 is fastened to extension6 of bearing blocks 75 by means of scre~s 81. By reference to Figures 7 and 13, it is seen that mounting plate 80 may be moved from it~ up~srd posit~on as sho~n in Figure 7 to its lo~er position as sho~n in Figure 13 by its attachment to bearing blocks 75 and their sliding engagement on posts 72.
A barrel bar 88 i9 secured to mounting plate 80 by means of ~cre~s 89. Mounted on barrel bar 88 are a .

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plurality of pipette barrels 92 having their heads 193 secured ~ithin the barrel bar 88 in a manner to be described below. As illustrated in the partial cut a~ay of barrel bar 88, the barrel lock bar 91 secures the lower portions of the barrels 92 to provide stability to the barrels. Guide tips 97 include adjustable screws 95 extending below the bottom edge of the mounting plate 80 which cooperate ~ith the lower surfaces 49 of plate 16 to accurately vertically position the lower tips 93 of barrels 92 with respect to the support medium 20 and blotting paper 30 disposed on lower surface 49 of plate 16. Such adjustment allows the droplets which form on the ends of the tips 93, when plungers 94 are driven do~nwardly within barrels 92, to "kiss" or be slightly appl$ed either to the support medium or the blotting paper. The droplets on the lower tips 93 of the barrels 92 are held because of their small size (as small as one micro liter) and surface tension forces of the barrel tips. When the tips are brought to a small distance within the upper surface of the support medium 20 or blotting paper 30, the droplets are relieved of the surface tension holding them to their barrels and are precisely applied to the blotting paper or to the support medium.
Actuator guides 183 are secured to the mounting plate 80 and include grooves in which an actuator plate 84 is inserted for sliding movement upwardly and downwardly with lZ~63B9 respect to the mounting plate 80. The actuator plate 84 has grooves in ~hich a plunger bar 90 is inserted. The plungers 94 of the microsyringe barrels 92 are secured to the plunger bar 90 and extend ~ithin the barrels 92. As illustrated in Figure 7, the plungers 94 are at their uppermost extent with respect to the barrels 92. The actuator plate 84 is adapted to move do~n~ardly with respect to the mounting plate 80, and through such action, the plunger bar 90 moves do~nwardly ~ith respect to the barrel bar 88 causing plungers 94 to move do~nwardly within the barrels 92 thereby forcing any fluid ~ithin such barrel~ out~ardly through the tips 93 of the barrels and forming a droplet at the tips of the barrels.
Position B~ gnals are generated indicative of the position of the mounting plate 80 with respect to the base 12 and the position of the actuator plate 84 and its plungers 94 ~ith respect to the mounting plate 80. The trip s~itch 106 mounted on the mounting plate 80 cooperates with the lower stop 115 mounted on the mounting block 76 to provide a lo~er mounting plate position signal when the mounting plate 80 reaches its lo~er extent. In a similar way as shown on the right hand side of the mounting plate 80, the upper trip switch 105 is mounted on the mounting plate 80 and is shown tripped by contact ~ith the upper stop 114. The upper trip s~itch 105 when tripped provides a signal to the microprocessor of lZ~63~9 elsctronics module 200 (Figure 3) indicative that the mounting plate 80 is in its upper position.
The trip switch 108 mounted on left hand side actuator 183 cooperates ~ith application position cam 100 and wash cam 102. The trip switch 108 is tripped by the application cam 100 when the actuator plate 84 moves upwardly with respect to the mounting plate 80 ana the trip switch 108 is tripped by wash cam 102 as the actuator plate 84 moves further upwardly. The do~n cam 104 trips trip ~witch 110 mounted on the right hand side actuator guide 183 when the actuator plate 84 reaches its maximum downward travel where the plungers 94 are within the barrels 92.
Figurè 9 is a cross-sectional view taken along lines 9-9 of Figure 7 and illustrates the mounting plate 80 and the actuator plate 84 both in their upward positions.
Mounting plate motor 116 has driven the mounting plate 80 to the upward position by operation of pinion 120 on rack 122 which is secured to the back plate 86 and to the base of the apparatus.
Turning no~ to Figure 10, the pipette assembly 70 is shown with the unting plate 80 in the downward position but the plunger bar 90 and the plungers 94 are in their upward position ~ith respect to the barrel bar 88. The barrels 92 are in a do~nward position in the wash well for aspirating 5 ~1 fluid, for example, from the ~ash well 26.

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Figure 11, a vie~ of the pipette assembly looking down~ardly along lines 11~11 from Figure 10, illustrates the drive mechanism by which the mounting plate 80 is moved up and down ~ith respect to the base. A mounting plate actuator motor 116 is fixed to the mounting plate 80 by means of a mounting screw 117. The output shaft of the motor 116 has a gear 119 fixed to it. Gear 119 is engaged with a left pinion gear 120L ~hich is mechanically coupled to a right pinion gear 120R by meanæ of shaft 124. The shaft is mounted to the mounting plate 80 by shaft mountings 118. Racks 122, fixed to the back plate 86, have their gears in engagement with pinion gears 120L and 120R. As the motor 116 turns in either direction under microprocessor control, the mounting plate 80 is moved upwardly or downwardly with respect to the base and the back plate 86 by the rack and pinion mechanism. The front view (Figure lO) of the pipette assembly 70 with the mounting plate 80 in a downward position shows the racks 122 visible. The front view also illustrates, with the mounting plate 80 in its maximum downward position, that stop scre~s 95 of guide tips 97 are slightly above surfaces 49 of the plate 16 indicative that tips 93 are ~lightly above the top edge 49 of the plate so that droplets which form on the tips may l'kiss" the surface 49 and apply sample fluid to the support medium or blotting paper.

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12~3~9~

Figure 12 is a cross-sectional view of the pipette apparatus taken along lines 12-12 of Figure 10 and sho~s that the mounting plate actuator motor 116 has turned its pinion gear 120 so that the mounting plate 80 has been moved do~nwardly with respect to the base and the raised portion 22 of the plate 16. Thus, the barrel 92 has been lowered to be within a well of the raised portion 22 of the sample plate. A wash well 26 is illustrated as an example ~here the barrels 92 of the pipettes have been lo~ered by the mounting plate 80 and where the fluid from the wash ~ells have been aspirated into the barrels 92 by virtue of the plungers 94 being pulled upwardly by means of the actuator plate 84. It is apparent from Figures 11 and 12 that the mounting plate 80 is translated upwardly and downwardly ~ith respect to the raised portion 22 by means of the motor 116 turning and causing the pinion 120 to translate upwardly and down~ardly on fixed rack 122.
Turning no~ to Figure 13, the state of the pipette apparatus 70 iB such that the actuator plate 84 has moved do~n~ardly causing the plungers 94 to be inserted back into the barrels 92 thereby positively displacing any fluid ~h$ch has been aspirated ~ithin the barrels either to an application space, a blotter, or to a ~aste ~ell.
It is seen that the trip switch 108 has been returned to a condition such that any upward movement of the actuator plate 84 will be tripped first by the application cam 100 and then th- wDsh cam 102 provlding D meanY for signa11ing ,' the position of the actuator plate 84 ~ith respect to the mounting plate 80.
Figure 14, is a downward looking vie~ along lines 14-14 of Figure 13 and illustrates the drive mechanism by which the actuator plate 84 is translated up~ardly and downwardly with respect to the mounting plate 80. A
plunger bar actuator plate motor 126 is fixed to the mounting plate 80 by means of a mounting scre~ 127. The motor 126 includes a gear 128 on its output shaft which is in engagement with pinion gear 130R. Pinion gear 130R is coupled to a pinion gear 130L by means of a shaft 134 which i~ supported by means of shaft mountings 118 which also supports shaft 124 (see Figure 11). The actuator plate 84 has actuator plate racks 132 fixed to the rear side thereof which extend through slots 136 in the mounting plate so as to engage the pini~ns 130L and 130R.
As the plunger bar actuator plate motor 126 i9 caused to turn in either the clock~ise or the counterclockwise direction, the actuator plate 84 is caused to move upwardly or do~n~ardly with respect to the mounting plate 80. Figure 13 shows the slots 136 in the mounting plate 80 through which the actuator plate racks 132 extend.
Figure 14 also shows the means by which the barrel heads 193 of the barrels 92 are removably fixed to the barrel bar 88. The barrel bar 88 comprises a receiving bar 138 having slots 140 provided along its front face.
The barrel heads 193 are inserted therein and secured by ~2~;3~39 means of a securing bar 139 which holds the barrels vertically in place. The securing bar 139 is secured to the receiving bar 138 by means of scre~s 141. The barrel lock bar 91 is similarly constructed as the barrel bar 88.
The barrel bar 88 provida a removably securin~ means by ~hich the barrels 92 may be easily replaced due to ~ear or breakage.
Figure 15 is a cross-sectional vie~ looking along lines 15-15 of Figure 13 and illustrates the mounting plate 80 in a do~nward pocition. $he actuator plate 84 has been tran~lated downwardly ~here the plunger bar 90 is adjacent the barrel bar 88. Figure 15 illustrates the actuator plate rackc 132 extending through slots of the mounting plate 80 and their engagement with pinion gear 130 ~hich has been turned by means of the actuator plate mntor 126. The barrel 92 is now in a rinse ~ell 28, for example. The plunger 94 has been forced do~n by means of the actuator plate 84 moving the plunger bar 90 to its lo~ermost position. Of course, the ra$sed portion of the plate 22 has moved longitudinally wlth respect to the - pipette assembly bet~een the vie~s of Figures 12 and lS.
Figure 16 illustrates schematically the means by which the carriage mounting plate and plunger actuator bars are controlled to perform the automatic pipetting operation. The dotted box 200 represents a microcomputer integrated circuit device, preferably a microcircuit No.
, HD68POlV07 manufactured by the Hitachi Corporation. The '~,' ~ ' ~ ' . - . .
. .
-l~iG389 circuit includes a central processing unit 201A, a read only memory 201B, a random access memory 201C, a timer 201D, an output interface circuit 201E and input interface circuit 201F. The read only memory circuit 201B includes stored soft~are by which the entire automatic operation is controllea and will be discussed below.
Figure 16 illustrates the carriage motor 40, the mounting plate motor 116 and the actuating plate motor 126 all under computer control via the motor driver circuits 202, 203, 204 such a6 circuits UDN-2952B manufactured by the SPRAGUE Corporation. These motor driver circuits are u6ed to control the speed of the motor and its direction of rotation. Also provided in conjunction ~ith the motor are electronic motor break circuits 205, 206, 207 which are provided to quickly break the motor's rotation on receipt of a translation signal by the computer 200. Such electronic motor break circuits are preferably 2N6075 Triac circuits.
The po6ition detector circuits 208 represent the circuitry with the trip switches 60 and 62 illustrated in Figure 5 which indicate the position of the carriage 46 and the sample plate 16 ~ith respect to the pipette assembly .
The position detector circuits 209 represent the circuitry associated with the lower trip switch 106 and the upper trip s~itch 105 ~hich signal the up~ard or lZf~38~

do~n~ard limits of travel of the mounting plate 80 ~ith respect to the base.
The position detector circuits 210 represent the circuitry associated ~ith trip s~itches 108 and 110 ~hich indicate the relative position of the actuator plate 84 ~ith respect to the mounting plate 80. The signals associated ~ith each of those position detector circuits are represented as bei~g carried by a bundle of electrical leads 215 to the input interface circuitry 201F of 10 electronic module 200.
The alarm circuit 211 i8 provided for the apparatus, for example, such as a sounding device EAF14R06C
manufactured by Panasonic. Such circuit is act~vated and a sound is generated to signal faults in the operation of the apparatus or to signal the readiness of the machine.
Indicator circuit 212 represents an indicator lamp as illustrated in Figure 5 to signal the user that the po~er is on to the apparatus. The interlock circuit 213 represents the circuitry ~ith sample plate interlock trip 20 s~itch 65 ~hich indicates the presence or absence of the sample plate on the carriage. Command circuit 214 represents a push button s~itch used to start or abort the automatic pipetting application process.
In operation, the central processing unit 201A
receives the sequence of events instructions from the s programs stored in the read only memory 20lB. The central processing unit 201A then receives positional information ', ., .
,, ,~ , , .

3~9 concerning the moving mechanisms of the apparatus by means of reading and decoding the binary coded data present at the input interface 201F which receives information via leads 215 from the position detector circuits 208, 209, 210.
The microproces~or CPU 201A then receives an input command to start or abort the process by means of reading and decoding the binary coded data present at the input interface 201F which is connected to the command circuit 214 whlch may be the push button 214 illustrated in Figure 5.
Figure 17 illustrates in flo~ chart form the operations of the CPU 201A under program control. The CPU
201A determines the validity of a command to start the processing by means of reading and decoding the binary coded data pre~ent at the input interface 201F which receives a signal from the interlock circuit 213. This operation insures that the plate 16 i8 fully inserted into ; the carriage.
The CPU 201A causes the motors 40, 116 or 126 to turn in the required direction be means of ~riting the appropriate binary coded data to the output interface circuit 201E which is connected to the motor drive circuit~ 202, 203 and 204. The microprocessor CPU 201A
then causes the mechanism movement to stop precisely when the re~uired location is reached by writing the appropriate binary coded data to the output interface , . . ..... ..

lZ~i3~

circuit 201E ~hich is connected to the motor drive circuits 202, 203, 204 to aisable the drive and then ~riting the appropriate binary coded data to the output interface 201E ~hich is connected to the motor break circuits 205, 206, 207 to apply electronic breaking.
The microprocessor circuit 201A then signals that the pipetting apparatus is ready or that a plate has been completed or that a failure has occurred by means of writing the appropriate binary coded data to the output interface circuit 201E connected to the alarm circuit 211 to sound an alarm.
The timer 201D of Figure 16 is used by the m$croproce~sor CPU 201A to determine eleatrical or mechanical failures of the positioning mechanism. This is accomplished by means of measuring the elapsed time during a command to drive any motor. If the event is not completed ~ithin the prescribed length of time, the drive command i6 aborted and the alarm is activated by means of the microproce~or CPU 201A writing the appropriate binary coded data to the output interface connected to the alarm circuit 211. The timer 201D i8 also used to determine the repetition rate of the alarm thereby allowing the microprocessor circuit CPU 201A to encript and communicate to the operator the nature of the failure.
As sho~n in Figure 17, once the automatic pipetting apparatus of the invention is running, a ~ide variety of ... .

dlffer-nt applic-tionc may be achieved. The ~equence of ' ~.
.', lZt~638~

operations sho~n in Figure 17 is preerred in that first, five microliters of cleansing agent such as distilled water is aspirated into the barrels of the pipettes from the wash well. Next, the water in the barrels is dispensed into the waste well. Then the applicator tips are dried by lowering them to a blotter pad as illustrated in Figure 5.
Next, the barrels are moved to their upward position with the plungers in their do~nward position, the carriage 0 i5 moved rearward and the barrels are lowered into the sample chambers 24. The plungers are raissd thereby dra~ing a small amount of each sample of liquid, for example, patient blood to be tested. Where no dilution of the blood samples is desired, the samples are applied precisely to the cellulose acetate or agarose strip. The barrels are raised again. Again, the carriage is moved forward until the wash well is beneath the barrels and the mounting plate is lo~ered such that distilled ~ater is again dra~n into the barrels and then dispensed into the ~0 waste well.
Where dilution of the sample liquids is desired, the sample plate of Figure 2B may be substituted for that of Figure 2 and the computer program illustrated by the flo~
chart of Figure 17 branches to the dilute routine. A
preferred routine for diluting the samples is to dra~ an additional four micro liters of fluid from the wash well.
This action results in each of the barrels being filled :
: , .

~2~63f~9 ~ith four micro liters of diluting fluid (e.g, ~ater) and one micro liter of blood (or other liquid) sample. Next the entire five micro liters of fluid of each barrel is applied to the respective dilution chambers of the dilution ro~ of the plate of Figure 2B. This process may be repeated a desired number of times to effect mixing of the sample ~ith the dilution fluid (~ater). Finally a one micro liter sample of the diluted blood sample is drawn into each barrel according to the description presented previously. The routine then proceeds as described above ~here the one micro liter of diluted liquid sample is applied to the support medium.
The dilution routine described above is preferred, but other routines may be used to effect good mixing of the blood sample with diluting liquid. For example, a first predetermined amount of liquid sample in each barrel may be applied to the dilution ~ells. A predetermined amount of ~ash liquid may then be applied to the dilution ~ells. After mixing the combination of the ~ash liquid and liquid sample of the dilution ~ells, (for example by the mixing technique described above), a small amount of liquid samples is aspirated from the dilution ~ells and applied to the support medium.
An appendix to this specification is filed herewith and the appendix is to be retained on the Patent file and includes a source listing of the computer program written in HD68POlV07 Hitachi Assembly language which is stored in the read only memory 201B so as to automatically control the pipetting, 2 ~ 3 cleansing, blotting, diluting ~at the operator's option) and other functions described above.
Various modifications and alterations in the described structures ~ill be apparent to those skilled in the art of the foregoing description ~hich does not depart from the spirit of the invention. For this reason, these changes are desired to be included in the appended claims.
The appended claims recite the only limitation to the present invention and the descriptive manner ~hich is employed for setting forth the embodiments and is to be interpreted as illustrative and not limitative.

12~36389 _ APPENDIX TO SPECIF1 ~TION
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Claims (36)

1. Automatic pipetting apparatus comprising a base, sample plate means disposed on said base, said plate means including a row of liquid sample chambers and a lateral application space longitudinally separated from said liquid chamber row, said lateral application space adapted to receive a microporous support medium, said sample chambers adapted to receive liquid samples, a pipette frame including vertical support means for supporting said frame from said base above said sample plate means, longitudinal translation means for effecting relative longitudinal movement of said pipette frame and said sample plate means, a mounting plate carried by said pipette frame, vertical translation means for effecting relative vertical movement of said mounting plate and said sample plate means, a plurality of microsyringe barrels having their heads secured in a row to said mounting plate, said barrels spaced corresponding to the spacing of said liquid chambers on said plate, said barrels being hollow with each having a lower tip, a plurality of micro-plungers, each of said plungers disposed in one of said barrels, plunger translation means for moving said plungers vertically within said barrels, signalling means for generating longitudinal signals representative of the relative longitudinal orientation of said pipette frame with respect to said sample plate means, for generating mounting plate signals representative of the vertical orientation of said mounting plate relative to said sample plate means and for generating plunger signals representative of the orientation of said plungers relative to said barrels, and programmed computer means responsive to said longitudinal signals, to said mounting plate signals and to said plunger signals for generating a sequence of control signals to said longitudinal translation means, to said vertical translation means and to said plunger translation means to aspirate a first predetermined amount of liquid from said sample chambers into said respective pipette barrels, and to apply said liquid samples in each of said pipette barrels, where said lower tips of said barrels are at a position slightly above said microporous support medium onto corresponding spaces of said microporous support medium when placed on said lateral application space of said sample plate.

2. The apparatus of claim 1 wherein said sample plate means is translated longitudinally beneath said pipette frame by said longitudinal translation means.

3. The apparatus of claim 1 wherein said pipette frame is translated longitudinally above said sample plate means by said longitudinal translation means.

4. The apparatus of claim 1 wherein said sample plate includes a wash well adapted to contain wash liquid and a waste well longitudinally spaced from each other and from said sample chambers, said programmed computer means generating a further sequence of control signals to said longitudinal translation means, to said vertical translation means and to said plunger translation means before aspirating liquid from said sample chambers, to draw a second predetermined amount of wash liquid from said wash well into respective pipette barrels, and to discharge said wash liquid into said waste well.

5. The apparatus of claim 4 wherein said sample plate means includes a longitudinal blotting space for applying a lateral blotting paper strip, said blotting space longitudinally separated from said sample chambers row, said wash well, said waste well and said lateral application space, and said programmed computer means generating a further sequence of control signals to said longitudinal translation means, to said vertical translation means and to said plunger translation means after discharging said rinse liquid into said waste well, to blot the tips of said barrels on said blotting paper strip.

6. The apparatus of claim 4 wherein said sample plate means further includes a row of individual liquid dilution wells longitudinally spaced from said sample chambers, said dilution wells adapted to receive dilution liquid, said programmed computer means generating a further sequence of control signals to said longitudinal translation means, to said vertical translation means and to said plunger translation means for applying said first predetermined amount of liquid sample in each barrel to said dilution wells, applying a third predetermined amount of wash liquid from each barrel to said dilution wells, mixing the combination of said wash liquid and said liquid sample of said dilution wells, and aspirating an amount of diluted liquid samples from said dilution wells into said respective pipette barrels.

7. The apparatus of claim 1 wherein said sample plate means further includes a wash well adapted to contain wash liquid and a waste well longitudinally spaced from each other and from said sample chambers and a row of individual liquid dilution wells longitudinally spaced from said sample chambers, said dilution wells adapted to receive dilution liquid, said programmed computer means generating a further sequence of control signals to said longitudinal translation means, to said vertical translation means and to said plunger translation means after aspirating liquid samples of said liquid samples into said pipette barrels, to aspirate a second predetermined amount of wash fluid from said wash well into said respective pipette barrels operably creating a combination of sample liquid and wash liquid in each barrel, to discharge said sample liquid/wash liquid into said corresponding dilution wells operably creating diluted liquid samples, and to aspirate a third predetermined amount of diluted liquid samples from said dilution wells into said respective pipette barrels.

8. The apparatus of claim 7 wherein:
said programmed computer means generates a further sequence of control signals to said plunger translation means to alternatingly discharge said sample liquid/wash liquid into said corresponding dilution wells and aspirate a predetermined amount of diluted liquid from said dilution wells into said respective pipette barrels operatively causing mixing of said liquid samples with said dilution liquid in said dilution wells.

9. A sample plate adapted for use with automatic pipetting apparatus comprising a lateral row of individual liquid sample chambers and a lateral application space longitudinally separated from said liquid chamber row, said lateral application space adapted to receive a microporous support medium and including a raised portion and a lower portion, said row of individual liquid sample chambers being disposed on said raised portion, said lateral application space being disposed on said lower portion and further comprising a wash well and a waste well longitudinally spaced from each other and from said sample chambers.

10. The sample plate of claim 9 further comprising a row of individual liquid dilution wells longitudinally spaced from said sample chambers.

11. A sample plate adapted for use with automatic pipetting apparatus comprising a lateral row of individual liquid sample chambers and a lateral application space longitudinally separated from said liquid chamber row, said lateral application space adapted to receive a microporous support medium, and further comprising a wash well and a waste well longitudinally spaced from each other and from sample chambers;
and further including a longitudinal blotting space for applying a lateral blotting paper strip, said blotting space longitudinally separated from said sample chamber row, said wash well, said waste well and said lateral application space.

12. The sample plate of claim 11 including a raised portion and a lower portion, said row of individual liquid sample chambers being disposed on said raised portion, said lateral application space being disposed on said lower portion.

13. The sample plate of claim 11 including a raised portion and a lower portion, said row of individual liquid sample chambers, said wash well and said waste well being disposed on said raised portion, said lateral application space and said blotting space being disposed on said lower portion.

14. Apparatus adapted for use with automatic pipetting of liquid samples to be subjected to electrophoresis comprising a sample plate including a lateral row of individual liquid sample chambers and a lateral application space longitudinally separated from said liquid chamber row, said lateral application space having disposed thereon a removable microporous support medium adapted for electrophoretic analysis after a liquid sample has been deposited thereon, said sample plate including an upper portion and a lower portion, said row of individual liquid sample chambers being disposed on said raised portion, said lateral application space being disposed on said lower portion.

15. Automatic pipetting apparatus comprising, a base, track means disposed longitudinally on said base, a carriage longitudinally movably disposed on said track means, a sample plate removably disposed on said carriage, said sample plate including a lateral row of individual liquid sample chambers and a lateral application space longitudinally separated from said liquid chamber row, said lateral application space adapted to receive a microporous support medium, said sample chambers adapted to receive sample liquids, a pipette frame mounted vertically on said base above said carriage and said sample plate, said pipette frame having vertical mounting posts separated longitudinally from each other, each of said posts secured to said base, a mounting plate assembly slidably guided by said posts and disposed laterally with respect to said sample plate and including, a mounting plate having slidable guides disposed about said posts, a barrel bar fixed to said mounting plate, a plurality of microsyringe barrels having their heads secured in a row in said barrel bar and spaced corresponding to the spacing of said liquid chambers on said plate, said barrels being hollow with each having a lower tip, a plunger bar vertically movably disposed above said barrel bar and having a plurality of micro-plungers secured thereto, each of said micro-plungers movably disposed within a corresponding barrel of said microsyringes, and a plunger actuator plate vertically movable with respect to said mounting plate and carried by said mounting plate, said actuator plate removably secured to said plunger bar, translation and signalling means for moving said carriage longitudinally forward beneath said mounting plate assembly and generating carriage position signals indicative of said carriage position, moving said mounting plate assembly vertically with respect to said base and generating mounting plate position signals indicative of mounting plate position, and moving said plunger actuator bar and said plunger bar vertically with respect to said mounting plate and generating plunger bar position signals indicative of plunger bar position, and programmed computer means responsive to said carriage position signals, to said mounting plate position signals and to said plunger bar position signals for generating a sequence of control signals to said translation means for translating said carriage, said mounting plate and said plunger bar to aspirate a first predetermined amount of liquid from said sample chambers into said respective pipette barrels, and to apply said liquid in each of said pipette barrels from a position slightly above said microporous support medium whereby droplets of said liquid which forms on the tops of said barrels are transferred from said tips onto corresponding spaces of said microporous support medium when placed on said lateral application space of said sample plate.

16. The apparatus of claim 15 wherein said sample plate includes a wash well and a waste well longitudinally spaced from each other and from said sample chambers, said programmed computer means generating a further sequence of control signals to said translation means before aspirating liquid from said sample chambers, to draw a second predetermined amount of rinse liquid from said wash well into respective pipette barrels, and to discharge said rinse liquid into said waste well.

17. The apparatus of claim 16 wherein said sample plate includes a longitudinal blotting space for applying a lateral blotting paper strip, said blotting space longitudinally separated from said sample chamber row, said wash well, said waste well and said lateral application space, and said programmed computer means generating a further sequence of control signals to said translation means after discharging said rinse liquid into said waste well, to blot the tips of said barrels on said blotting paper strip.

18. The apparatus of claim 17 wherein said sample plate includes a raised portion and a lower portion, said row of individual liquid sample chambers being disposed on said raised portion, said lateral application space being disposed on said lower portion.

19. The apparatus of claim 17 wherein said sample plate includes a raised portion and a lower portion, said row of individual liquid sample chambers, said wash well and said waste well being disposed on said raised portion, said lateral application space and said blotting space being disposed on said lower portion.

20. The apparatus of claim 15 wherein said translation and signalling means comprises, carriage translation means responsive to carriage translative signals for longitudinally translating said carriage and said sample plate on said track means beneath said pipette assembly for operably translating said plate to at least a sample chamber position where said sample chambers are beneath said microsyringe barrels and to an application position where said microporous support medium is beneath said microsyringe barrels, carriage signalling means for generating a sample chamber carriage position signal when said sample chambers are beneath said microsyringe barrels, and an application position signal where said microporous support medium is beneath said microsyringe barrels, mounting plate assembly translating means responsive to up and down mounting plate assembly translation signals for vertically translating said mounting plate assembly between an upper position and a lower position, said upper position being sufficiently high to lift said microsyringe barrels above said plate operably allowing translation of said plate beneath said barrels, said lower position operably allowing insertion of said barrels into said fluid sample chambers when said plate is in said sample chamber position and operably allowing said barrels to be slightly above said microporous support medium when secured to said lateral application space of said plate, mounting plate signalling means for generating a mounting plate upper position signal when said mounting plate is in said upper position and a mounting plate lower position signal when said mounting plate is in said lower position, plunger actuator plate translating means responsive to up and down plunger actuator plate translation signals for vertically translating said plunger actuator plate and said plunger bar between a lower position where said micro-plungers are fully within said corresponding microsyringe barrels to a sample upper position where said micro-plungers are extended upwardly through said barrels operably drawing fluid into said barrels when said tips are immersed in fluid in said sample chamber, plunger actuator plate signalling means for generating a plunger bar lower position signal when said plunger actuator plate and said plunger bar are in said lower position, and a plunger bar sample upper position signal when said plunger actuator plate and said plunger bar are in said sample upper position, and electrical means for providing signal communication between said programmed digital computer and said carriage translation means, said mounting plate assembly translating means, and said plunger actuator plate translating means, and for providing signal communication between said digital computer and said carriage signalling means, said mounting plate signalling means, and said plunger actuator plate signalling means.

21. The apparatus of claim 20 wherein said digital computer means includes a stored sequence of instructions for generating and applying said carriage translation signal to said carriage translation means thereby translating said carriage to said sample chamber position and removing said carriage translation signal when said sample chamber carriage position signal is received, said programmed digital computer means operably generating and applying said down mounting plate assembly translation signal to said mounting plate assembly translating means thereby translating said mounting plate assembly from said upper position to said lower position and removing said mounting plate assembly translation signal when said mounting plate lower position signal is received, said programmed digital computer means operably generating and applying said up plunger actuator plate translation signal to said plunger actuator plate translating means thereby translating said plungers from said lower position to said sample upper position and removing said plunger actuator plate translation signal when said plunger bar sample upper position signal is received, operably causing fluid in each of said sample chambers to be aspirated into said corresponding barrels, said programmed digital computer means operably generating and applying said up mounting plate assembly translation signal to said mounting plate assembly translating means thereby translating said mounting plate assembly from said lower position to said upper position and removing said mounting plate assembly translation signal when said mounting plate upper position signal is received, said programmed digital computer means operably generating and applying said carriage translation signal to said carriage translation means thereby translating said carriage to said application position and removing said carriage translation signal when said application carriage position signal is received, said programmed digital computer means operably generating and applying said down mounting plate assembly translation signal to said mounting plate assembly translating means thereby translating said mounting plate assembly from said upper position to said lower position and removing said mounting plate assembly translation signal when said mounting plate lower position signal is received, and said programmed digital computer means operably generating and applying said down plunger actuator plate translation signal to said plunger actuator plate translating means thereby translating said plunger from said sample upper position to said lower position and removing said plunger actuator plate translation signal when said plunger bar sample lower position signal is received, operably causing fluid in each of said barrels to be applied to said microporous support medium when secured to said lateral application space of said plate.

22. The apparatus of claim 21 wherein said sample plate further includes a wash well and a waste well longitudinally spaced from each other and from said sample chambers, and said plunger actuator plate translating means further includes a wash upper position, and said plunger actuator plate signalling means further includes means for generating a plunger bar wash upper position signal when said plunger actuator plate and said plunger bar are in said wash upper position, said carriage translation means further includes a wash well position and a rinse well position, and said carriage signalling means further includes means for generating a wash well carriage position signal when said wash well is beneath said microsyringe barrels and a waste well carriage position signal when said waste well is beneath said microsyringe barrels, said digital computer means includes a further stored sequence of instructions for before aspirating liquid from said sample chambers, generating and applying said carriage translation signal to said carriage translation means thereby translating said carriage to said wash well position and removing said carriage translation signal when said wash well carriage position signal is received, said programmed digital computer means operably generating and applying said down mounting plate assembly translation signal to said mounting plate assembly translating means thereby translating said mounting plate assembly from said upper position to said lower position and removing said mounting plate assembly translation signal when said mounting plate lower position signal is received, said programmed digital computer means operably generating and applying said up plunger actuator plate translation signal to said plunger actuator plate translating means thereby translating said plungers from said lower position to said wash upper position and removing said plunger actuator plate translation signal when said plunger bar wash upper position signal is received, operably causing fluid in said wash well to be aspirated into said barrels, said programmed digital computer means operably generating and applying said up mounting plate assembly translation signal to said mounting plate assembly translating means thereby translating said mounting plate assembly from said lower position to said upper position and removing said mounting plate assembly translation signal when said mounting plate upper position signal is received, said programmed digital computer means operably generating and applying said carriage translation signal to said carriage translation means thereby translating said carriage to said wash well position and removing said carriage translation signal when said wash well carriage position signal is received, said programmed digital computer means operably generating and applying said down mounting plate assembly translation signal to said mounting plate assembly translating means thereby translating said mounting plate assembly from said upper position to said lower position and removing said mounting plate assembly translation signal when said mounting plate lower position signal is received, said programmed digital computer means operably generating and applying said down plunger actuator plate translation signal to said plunger actuator plate translating means thereby translating said plunger from said wash upper position to said lower position and removing said plunger actuator plate translation signal when said plunger bar lower position signal is received, operably causing fluid in each of said barrels to be applied to said waste well, and said programmed digital computer means operably generating and applying said up mounting plate assembly translation signal to said mounting plate assembly translating means thereby translating said mounting plate assembly from said lower position to said upper position and removing said mounting plate assembly translation signal when said mounting plate upper position signal is received.

23. The apparatus of claim 22 wherein said sample plate further includes a blotting space for applying a lateral blotting paper strip, said blotting space longitudinally separated from said sample chamber row, said wash well, said waste well and said lateral application space, said carriage translation means further includes a blotting position, and said carriage signalling means further includes means for generating a blotting carriage position signal when said blotting space is beneath said microsyringe barrels, said digital computer means includes a further stored sequence of instructions for after discharging said rinse liquid to said waste well, generating and applying said carriage translation signal to said carriage translation means thereby translating said carriage to said blotting position and removing said carriage translation signal when said blotting carriage position is received, said programmed digital computer means operably generating and applying said down mounting plate assembly translation signal to said mounting plate assembly translating means thereby translating said mounting plate assembly from said upper position to said lower position and removing said mounting plate assembly translation signal when said mounting plate lower position signal is received, whereby said barrel tips are blotted when said blotting paper is disposed on said blotting space, and said digital computer means includes a further stored sequence of instructions for said programmed digital computer means operably generating and applying said up mounting plate assembly translation signal to said mounting plate assembly translating means thereby translating said mounting plate assembly from said lower position to said upper position and removing said mounting plate assembly translation signal when said mounting plate upper position signal is received.

24. The apparatus of claim 15 wherein said means for translating said carriage longitudinally forward and backward beneath said mounting plate assembly comprises a carriage actuating motor fixed to said base, said motor having a rotatable output shaft, a pinion gear fixed to said output shaft, a rack gear secured to said carriage, said rack gear being in engagement with said pinion gear whereby the output shaft turning of said motor translates said carriage.

25. The apparatus of claim 18 wherein said signalling means for generating carriage position signals indicative of carriage position includes said carriage having a first longitudinal surface with notches disposed thereon, the longitudinal separation of said notches corresponding to the longitudinal position of said wash well, said waste well, said longitudinal blotting space, said sample chambers and said lateral application space, a first trip switch secured to said track means on which said carriage moves, said switch having a first spring forced roller means for engaging said longitudinal surface whereby said switch is tripped when said roller is forced into a notch, operatively indicating that said wash well or said waste well or said longitudinal blotting space, or said sample chambers, or said lateral application space is beneath said microsyringe barrels.

26. The apparatus of claim 25 wherein said signalling means for generating carriage position signals indicative of carriage position includes said carriage having a second longitudinal surface with at least one notch disposed thereon near the forward end of said carriage, a second trip switch secured to said track on which said carriage moves, said second switch having a second spring forced roller means for engaging said second longitudinal surface whereby said switch is tripped when said second roller means is forced into said notch operatively indicating that said carriage has reached its rearward limit of travel.

27. The apparatus of claim 26 further comprising a limit switch means secured to the rear of said carriage at a position for engaging the forward end of said plate for indicating that said plate is in an operative position within said carriage.

28. The apparatus of claim 15 wherein said means for translating said mounting plate assembly up and down with respect to said base comprises a mounting plate actuator motor fixed to said mounting plate, said motor having a rotatable output shaft, pinion gear means fixed to said output shaft, a rack gear secured to said base, said rack gear being in engagement with said pinion gear means whereby the output shaft turning of said mounting plate actuator motor translates said mounting plate up or down with respect to said base.

29. The apparatus of claim 15 wherein said signalling means for generating mounting plate position signals indicative of mounting plate position includes upper stop means secured to a member secured to said base, upper trip switch means secured to said mounting plate and having a trip arm for engaging said upper stop means for operatively tripping said upper trip switch means when said mounting plate reaches its upper limit of travel, lower stop means secured to said base, and lower trip switch means secured to said mounting plate having a trip arm for engaging said lower stop means for operatively tripping said lower trip switch means when said mounting plate reaches its lower limit of travel.

30. The apparatus of claim 18 wherein said means for translating said plunger bar up and down with respect to said mounting plate comprises a plunger plate motor fixed to said mounting plate, said motor having a rotatable output shaft, pinion gear means fixed to said output shaft, rack gear means fixed to said plunger actuator plate, said rack gear being in engagement with said pinion gear means whereby the output shaft turning of said plunger actuator plate motor translates said actuator plate and said plunger bar up or down with respect to said mounting plate.

31. The apparatus of claim 30 wherein said plunger actuator plate motor is fixed to the rear side of said mounting plate, said mounting plate includes two vertically plunger actuator plate guides with vertical grooves provided therein, said guides being fixed to the forward side of said mounting plate, said plunger actuator plate is slidingly disposed within said vertical grooves on said mounting plate, said mounting plate has vertical slots therein, and said rack gear means fixed to said plunger actuator plate extend from said plunger actuator plate on the forward side of said mounting plate to said pinion gear means fixed to said output shaft of said plunger actuator plate motor on the rear side of said mounting plate.

32. The apparatus of claim 30 wherein said signalling means for generating plunger bar position signals indicative of plunger bar position includes application cam means vertically adjustable on a vertical shaft secured to said plunger actuator plate, wash cam means vertically adjustable on a vertical shaft secured to said plunger actuator plate, a first trip switch means secured to said mounting plate and having a trip arm for engaging said application cam means for operatively tripping said first trip switch means when said plunger actuator plate reaches an upper application position, and operatively tripping said first trip switch means when said plunger actuator plate reaches an upper wash position, lower cam means vertically adjustable on a vertical shaft secured to said plunger actuator plate, and a second trip switch means secured to said mounting plate and having a trip arm for engaging said down cam means for operatively tripping said second trip means when said plunger actuator plate reaches its lower limit of travel.

33. The apparatus of claim 15 wherein said sample plate means further includes a row of individual liquid dilution wells longitudinally spaced from said sample chambers, said diluted wells adapted to receive dilution liquid, said programmed computer means generating a further sequence of control signals to said longitudinal translation means, to said vertical translation means and to said translation signalling means for moving said plunger actuator bar and said plunger bar vertically with respect to said mounting plate for applying said first predetermined amount of liquid sample in each barrel to said dilution wells, applying a third predetermined amount of wash liquid from each barrel to said dilution wells, mixing the combination of said wash liquid and said liquid sample of said dilution wells, and aspirating an amount of diluted liquid samples from said dilution wells into said respective pipette barrels.

34. Automatic pipetting apparatus comprising a base, sample plate means stationarily disposed on said base, said plate means including a row of liquid sample chambers and a lateral application space longitudinally separated from said liquid chamber row, said lateral application space adapted to receive a microporous support medium, said sample chambers adapted to receive liquid samples, a pipette frame including vertical support means for supporting said frame from said base above said sample plate means, longitudinal translation means for effecting longitudinal movement of said pipette frame above said stationarily disposed sample plate means, a mounting plate carried by said pipette frame, vertical translation means for effecting relative vertical movement of said mounting plate and said sample plate means, a plurality of microsyringe barrels having their heads secured in a row to said mounting plate, said barrels spaced corresponding to the spacing of said liquid chambers on said plate, said barrels being hollow with each having a lower tip, a plurality of micro-plungers, each of said plungers disposed in one of said barrels, plunger translation means for moving said plungers vertically within said barrels, signalling means for generating longitudinal signals representative of the relative longitudinal orientation of said pipette frame with respect to said sample plate means, for generating mounting plate signals representative of the vertical orientation of said mounting plate relative to said sample plate means and for generating plunger signals representative of the orientation of said plungers relative to said barrels, and programmed computer means responsive to said longitudinal signals, to said mounting plate signals and to said plunger signals for generating a sequence of control signals to said longitudinal translation means, to said vertical translation means and to said plunger translation means to aspirate a first predetermined amount of liquid from said sample chambers into said respective pipette barrels, and to apply said liquid samples in each of said pipette barrels onto corresponding spaces of said microporous support medium when placed on said lateral application space of said sample plate.

35. The apparatus of claim 1, 2, 3, 4, 5, 6, 7 or 8 wherein the application of said liquid samples in each of said pipette barrels comprises applying a droplet of said liquid samples onto corresponding spaces of said microporous support medium when placed on said lateral application space of said sample plate by precisely positioning said lower tips of said barrels at a small distance above said microporous support medium, said small distance being smaller than the diameter of a droplet of liquid sample which may be maintained on the end of said tips through surface tension forces of the barrel tips, whereby each droplet slightly touches said microporous support medium and is thereby relieved of its surface tension and is precisely transferred to said microporous support medium from each of said pipette barrels.

36. The apparatus of claim 15, 16, 17, 18, 19, 20, 24, 25, 26, 27, 28, 29, 30, 31, 32, 33 or 34 wherein the application of said liquid samples in each of said pipette barrels comprises applying a droplet of said liquid samples onto corresponding spaces of said microporous support medium when placed on said lateral application space of said sample plate by precisely positioning said lower tips of said barrels at a small distance above said microporous support medium, said small distance being smaller than the diameter of a droplet of liquid sample which may be maintained on the end of said tips through surface tension forces of the barrel tips, whereby each droplet slightly touches said microporous support medium and is thereby relieved of its surface tension and is precisely transferred to said microporous support medium from each of said pipette barrels.