AU595022B2 - Automatic pipetting apparatus - Google Patents

Description

Signed .zP L.....Status,,rqqsdent of ,Helena ,Laboratories Corporation Declarant's Name -TIPTON GOLIAS CO0M MO0N WE ALTH OF AIS T RA L IA PATENT ACT 19 CO-Mr.DETE SPECIF1I kTION (original) FOR OFFICE US, Class Int. Class Application Number: Lodged: 7, A7kF7 Complete Specification Lodged: Accepted: Published: t Priority: Related Art: 59502 2 4

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Name of Applicant: Address of Applicant: Actual Inventor(s) Address for Service: HELENA LABORATORIES CORPORATION P.O. Box 752, Beaumont, Tex-as 77704-0752, United States of America ROBERT J. SAIRRINE HE~NRY A. GAR SE DAVIES COLLISON, Patent Attorneys, 1 Little Collins Street, Melbourne, 3000.

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a Complete Specification for tho invention entitled: !'AUTOMATIC PIPETTING APPARATUS" Thel following statement is a full description of this invention, incl,ding the best method of performing it known to us r TITLE: AUTOMATIC PIPETTING APPARATUS BACKGROUND OF THE INVENTION Field of the Invention This invention is related in general to the field of applying fluid samples to analysis strips. In particular, this invention relates to an automatic pipetting apparatus for applying multiple fluid samples to a microporous support medium such as a cellulose acetate or agarose strip which 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 in 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 o0 proteins.

o0. Description of the Prior Art In the electrophoresis technique, a blood or other a0 fluid sample is applied to a support medium which 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 la -2support medium such that each of the samples may be subjected to the electric field at the same time.

The samples may be applied to the support medium one at a time in serial fashion with a hand pipetter, but the hand pipetter must be rinsed with a cleansing agent and blotted before a new sample is aspirated and then applied to thr strip.

Applicators have been designed to apply fluid samples simultaneously or in "parallel" t! the strips. Such applicators are described at page 61 of the General Products Catalog for 1384-1985 of Helena Laboratories with offices in Beaumont, Texas. Such applicators may apply eight, twelve or more samples to a microporous support medium and have the advantage of making the electrophoresis technique easier and more reproducible.

The applicators known prior to this invention however have been essentially non-automatic applicators and required cleaning of the applicator tips after each application to the support medium.

20 Automatic dispensing systems are kinown in the prior art. For example, a system sold under the trademark "Well Washers" of BioTech Instruments, Inc. of Burlington, Vermont provides an alignment mechanism by which a row of eight or twelve barrels may be positioned above one of a plurality of rows of washing vials or wells. Automation in the system provides selection of dispensing fill volumes, soaking times and number of wash cycles.

None of the prior art however has provided an apparatus for automat:ically filling a plurality of pipettes 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 washing and cleaning the barrels during each cycle time so as to prevent contamination of each of the barrels during S" application of a new plurality of fluid samples to a new support medium.

o Another disadvantage of the prior art is that there has been no means for precisely o automatically applying a very small amount of the °order of one micro-litre 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 o° order of one micro-litre of sample fluid with a *oa diluting liquid, and precisely applying a very small amount of the diluted sample to a support medium.

SUMMARY OF THE INVENTION %o In accordance with the present invention there is provided an automatic pipetting apparatus comprising a base, sample plate means disposed on said base, said sample plate means having longitudinal and lateral dimensions, said plate means including a lateral row of liquid sample chambers and a lateral 3 -L i -yxi__ ^ll iT- I-iL.1 I f ill(.i- i.l :i 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 laterally above said sample plate means, said pipette frme having a longitudinal position with respect to said longitudinal dimension of said sample plate means, logitudinal translation means for changing Si'" the longitudinal position between said pipette frame and said sample plate means, a mounting plate carried by said pipette frame, vertical translation means for effecting t 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, Sor. 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 Sorientation of said pipette frame with respect to said sample plate means, for generating mounting plate signals representative of the vertical orientation of said :i

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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 geierating 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 a droplet of 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 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 S' diameter of a droplet of liquid sample which may be .r t 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 0 support medium from each of said pipette barrels.

In accordance with the present invention there is also provided an automatic pipetting apparatus comprising a base, sample plate means disposed on said base, said sample plate means having longitudinal and 6 lateral dimensions, said plate means including a lateral 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 laterally above said sample plate means, said pipette frame having a longitudinal position with respect to said longitudinal dimension of said sample plate means, longitudinal translation means for changing the relative longitudinal position between said 0 0 pipette frame and said simple 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 o< 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 ,j being hollow with each having a lower tip, a plurality of micro-plungers, each of said Splungers 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 'ith respect to 7 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 1 of control signals to said longitudinal translation means, to said vertical translation means and to said 444 0 Soplunger translation means to aspirate a first predetermined amount of liquid from Caid 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 04 support medium when placed on said lateral 00oL. application space of said sample plate, wherein said sample plate means 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 So* 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 ii t t 4 I tI t 4t it i;: *i #4 4£ 4r 4 4* 44*4 44 8 pipette barrels, and to discharge said wash liquid into said waste well, and 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, In accordance with the present invention there is also provided 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.

In accordance with the present invention there is

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1 4 0* 44 4 4 4 *1%r 4 4 *C6 044*Q 4* 0 4 04 O 9 also provided a sample plate adapted for use with automatic pipetting apparatus colprising 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 Leceive 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.

In accordance with the present invention there is also provided an 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 3 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 a a moving said carriage longitudinally forward beneath said mounting plate assembly and generating I, carriage position signals indicative of said carriage position, moving said mounting plate assembly vertically with respect to said base an? vating mounting plate posiion. tons ini~ai' mounting plate position, an4 moving a A ctuator bar and said '0 0* i 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 a droplet of said liquid samples in each of said pipette barrels onto corresponding spaces of said microporous support "U medium when placed on said lateral application space U' 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 mieroporous 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.

4 In accordance with the present invention there is also provided an automatic pipetting apparatus comprising, a base, track means disposed longitudinally on said base, -1IE~i_ ii 9 c 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 0: pipette frame having vertical mounting posts "o 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, *o 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 0 chambers on said plate, said barrels being hollow with esch having a lower tip, a plunger bar vertically movably disposed o. above said barrel bar and having a plurality of s 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 S"r O A -n 1 9d 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 Smoving 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 S, 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 Scarriage, 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 svipport medium whereby droplets of said °Aj" Iliquid which forms on the tops of said barrels are ,o transferred from said tips onto corresponding spaces of said microporous support medium when placed on said lateral application space of said sample plate, a rkd wherein said sample plate includes a wash well and a waste well longitudinally spaced from each other and from said sample chambers, 9 a said programmed computer means generating a further sequence of control signals to said translation means before aspirating liq,;id from said sample chambers, to draw a se,-nd predeterminied amount of rinse liquid from said wash well into respective pipette barrels, and to discharge said rinse liquid into said waste well and wherein said sample plane includes a V, longitudinal blotting space for applying a lateral S, blotting paper strip, said blotting space 4 r, 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.

In accordance with the present invention there is also provided an automatic pipetting apparatus 4 0 comprising, Sa 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 t itr tt t ii Ui 4 944$ 4 Blr 44 b* B 4 4 application space lateral application space adapted to receive a mn.croporous support medium, said sample chambers adapted to receive sample liquids, 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 secures to said base, a mounting plate assembly slidably guided by said posts and disposed laterally with :espect to said sample plate and including, a mounting plate having ebid by 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-plunigers secured thereto, each of said micro-plungers movably disposed within a corresponding barrel of said microsyringes. and a plunger actuator plate vertically movably 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, 1 p- 11

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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 form 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, 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 1 9h 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 micosyringe 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 S, medium when secured to said lateral application space Sof said plate, mounting plate signalling means for generating a mounting plate upper position signal o«o: when said mounting plate is in said upper position and a mounting plate lower position signal when said Smounting plate is in said lower position, r plunger actuator plate translating means responsive to up and down plunger a- uator 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 i 41~ :i~~tE to .4 940 *40 4 *-a.r 4 tt 10 a. 4* a *0 0*4.i 0 *000 0 4

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a 4r a 4 00 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, wherein said digita, 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 9j plate assembly translating means thereby translating said mounting plate asse:mbly 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 S 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 I to said ujpp3 position and removing said mounting Ii t plate assembly translation signal when said mounting .'II plate upper position signal is received, said programmed digital computer means opeabLy generating and applying said carriage translation signal to said carriage translation means t Ahereby 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 iio said mounting 9k plate assembly translating means thereby translating said mounting plate assemblv. from said upper position to said lower position and remonving said mounting plate assembly translation signal when said mounting plate lower position signal is received, and said programmed digital computer r;operably generating and applying said dow- l,,,.ner 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 tci said microporous support medium when secured to said lateral application space of said plate, wherein said sample plate further includes a wash well and a waste well longitudinally spaced from each other and from said sample chambers, Sand 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 S.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 1' beneath said 9 1 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 S• 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 a received, said programmed digital computer means operably generating and applying said up plunger actuator pl-te translation signal to said plunger actuator plate translating means thereby translating said plungers from said lower position to said wash r 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 -t ci-l i 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9m 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 operablyr generating and applying said down mounting plate assembly translation signal to 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 C operably generating and applying sa d down plunger actuator plate translation signal ,j said plunger so actuator plate translating means thereby translating said plunger from said wash upper position to said lower position and removing said plunger actuator t.,I 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 NrO

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9n 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, and 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, t t said carriage translation means further includes a blotting position, and said carriage signalling meant 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 t 4 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 1 9 o plate lower position signal is received, whereby said barrel tips are blotted when said blotting paper is disposed on said blotting space, 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.

In accordance with the present invention there is also provided an automatic pipetting apparatus comprising a base, sample plate means stationarily disposed on said base, said sample plate means having longitudinal and lateral dimensions, said plate means including a lateral 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, n* pipette frame including vertical support ;means for supporting said frame from said base laterally above said sample plate means, longitudinal translation means for effecting longitudinal movement of said pipette frame above said stationarily disposed sample plate means, |n S 1 -I IIIC1~ 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 S*representative of the vertical orientation of said mounting plate relative to said sample plate means and "o 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 I I 9q apply said liquid samples in each of said pipette barrels onto corresponding spaces of said micropo support medium when placed on said lateral application space of said sample plate.

Brief Description of the Drawings The 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 S* which: r* 4* 0* 4 44* 44 Figure 1 shows a perspective view 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 showing the functional relationship between the sample plate with its sample chambers, wash well, waste 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 down and a plunger bar secured to plungers which are moved up and down with 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 is a perspective view of another embodiment of the sample plate further including a row of dilution wells; Figure 3 is a side view partially cut away and in 46,0, 20 section with the cover removed and taken from the view along lines 3-3 of Figure 1; Figure 4 is a cross-section of the carriage and the sample plate; Figure 5 is a downward looking view along lines S of Figure 1 and shows the cross-section of the base at level 5-5 and the track, carriage and sample plates beneath tho pipetting assembly; -11- Figure 6 is oss-sectional view taken along lines 6-6 of Figure 5 snd shows the base, the carriage and sample plate according to the invention and further shows the translation and guiding means by which the track is moved forward and backward with respect to the pipetting assembly; Figure 7 is a forward looking view taken along lines 7-7 of Figure 3 and shows partially broken away, partially cross-sectional parts of the pipette assembly slidably mounted on posts secured to the base and including a mounting plate which moves up and down with respect to the base and carrying a row of barrels of individual pipettes and an actuator plate movable vertically with respect to os0 the mounting plate for moving a plunger bar vertically for 64 S moving individual plungers within the barrels of each of *0 s 4"0 the pipettes; Figure 8 shows a top view of the pipette assembly showing in dashed lines the mounting plate of the pipette assembly; 20 Figure 9 shows a cross-sectional view taken along lines 9-9 of Figure 7 and illustrates the relationship between the mounting plate, the tip bar and the plunger bar and the means by which the, mounting plate is moved up and down with respect to the base; Figure 10 is a similar view to that of Figure 7 but shows the mounting plate having been translated to a lower position but with the plunger bar remaining in an upward ii -12position whereby the tips of the pipetters are in a downward position, but the plungers are extended upwardly from each of the barrels of the pipettes; Figure 11 is a top cross-sectional view taken along lines 11-11 of Figure 10 and illustrates the motor and rack and pinion system by which the mounting plate is moved up and down with respect to the base; Figure 12 is a cross-sectional view taken along lines 12-12 of Figure 10 and illustrates the relationship of the mounting bar after it has been moved downwardly by the rack and pinion system by operation of the motor turning with respect to the frame mounted rack; Figure 13 is a view similar to that of Figures 7 and but illustrates the plunger actuator plate and plunger e bar moved downwardly with respect to the mounting plate Itt 4 thereby forcing the plungers associated with each of the o 4 pipetters into their barrels and forcing any fluid previously aspirated into the barrels out the tips of the barrels; 20 Figure 14 shows a cross-sectional view taken along lines 14-14 of Figure 13 and illustrates the rack and *4£ pinion system by which the actuator plate and the plunger bar secured thereto is moved up and down with respect to the mounting plate; r* •Figure 15 is a cross-sectional view taken along lines 15-15 of Figure 13 and illustrates the movement of the actuator plate downwardly with respect to the mounting I rr, /1 i -13plate by operation of the rack and pinion system controlling relative movement between the actuator plate and the mounting plate; Figure 16 is a schematic illustration of the microcomputer integrated circuit device receiving signals from position detector circuits associated with 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.

3l 9 49 DESCRIPTION OF THE INVENTION Figure 1 illustrates in a perspective view the automatic pipetting apparatus 10 according to the o 4 invention. The apparatus includes a base 12 on which a sample plate 16 is movably supported. The sample plate o o 'Re 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 which a pipette head 18 is shown.

i ;i -14- Figure 2 is a schematic illustration of the essential mechanical elements of one embodiment of the invention with the base and the cover removed. None of the mounting apparatus is shown 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 shown in a functional way rather than in actual mechanical detail which will be shown in detail in the figures and discussion below.

The sample plate 16 as shown includes a row of sample chambers 24 as well 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 well 28 and the wash well 26 are provided on raised portions 22 of the sample plate. If a 'o desired, a plastic cup may be provided in each of sample S" chambers 24. A blotter space 32 between the rinse well 28 and the sample chambers 24 is provided at substantially the same vertical level 49 as the sample application space 21 on which the support medium 20 is secured.

As illustrated in Figure 2, the various regions of the sample plate are longitudinally distinct, yet the j* lateral spacings between the saiple chambers 24 corresponds to the application spots 19 on the support 1 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

I

I I Ii( _liliiI _11 ~I those chambers is aspirated by the automatic pipetting apparatus and is applied in a similar row on the spots 19 of the microporous support medium It is advantageous to provide the sample chambers 24, waste well 28 and wash well 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 downward position during all wash, waste, blot, sampling and application operations. However, it would be obvious to one of ordinary skill in the art that other arrangements could be provided especially where different levels of the mounting plate could be provided in the translation and signalling apparatus for controlling the mounting plate A detailed discussion of such translation and t signalling apparatus for controlling the mounting plate e 1 t s discussed below.

t.

4* The schematic illustration of Figure 2 shows that the 0 sample plate 16 is translated in forward and rearward directions beneath the pipette assembly 70 by virtue of the tmotor 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 Turning now to the pipette assembly 70 shown in g Figure 2, a mounting plate 80 is translated upwardly and 44t downwardly by means of mounting plate motor 116 having its pinion 120 engaging a rack 122. Thus, the entire mounting 1 -16plate 80, and the microsyringe barrels 92 attached to the barrel bar 88 which is secured to the mounting plate 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 down with 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 shown 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.

0 Figure 2 therefore shows all of the essential elements as far as the translation of the sample plate 16 backward and forward beneath the microsyringe barrels 92 2 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 Sa the plunger bar 90 and actuator plate 84 with respect to the mounting plate Figure 2A schematically illustrates an alterna,.'ve embodiment of the invention where the sample plate remains immovable with respect to the base 12' with the pipette assembly 70' being mounted on rollers 300 for longitudinal translation. Figure 2A illustrates that the sample plate j i r -17may 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 16B includes a row of sample chambers 24', a waste well 28', a wash well 26' and a blotting space 32'. The operation of the alternative embodiment is similar to that of the embodiment of Figure 2 except that translation and signalling means are provided for longitudinally translating pipette assembly 70' with 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' which may include an additional row of dilution wells in addition to the row of sample chambers 24, waste well 28, wash well 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 view taken along lines 3-3 of Figure 1 with a portion of track 34 cut away 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 4* 4' 4 4*4 4 44 44 4 *re 444 4 o 4 4, gt 44 4 444C 44(i 4.O~ 4 *44 I* 44 44 4 4 4,4' .4 44 4 4444 4) 4 -18pipette 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 upward 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 engagement with rack gear 36.

Notches are provided along the left edge of the carriage 46. These notches cooperate with 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 P. r 3.

Figure 4 illustrates a vertical cross-section through the carriage 46 and the plate 16 and shows the actual wash a* well 26, rinse well 28 and one of the sample chambers 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 downward view taken along lines 5-5 of Figure 1. Carriage 46 is shown supported by tracks 34 and 1 ~i-n iL-- ;I lii-il I li -19movable in the rearward and forward directions by means of mo,.r 40, pinion 38 and rack 36 as also illustrated in Figure 6. The sample plate 16 is disposed in a valley or U-shaped cross-sectional structure 47 of carriage 46.

Position signalling notches on the sides of the carriage 46 cooperate with trip switch 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 past them. For example, the notches on the left hand side 46 include the wash notch 50, the waste notch 52, the blot notch 54, the sample chamber notch 56 and an application notch 58. The notches correspond to the longitudinal position of the wash well 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 where the wash well 26 is directly beneath the barrels 92, the roller 61 moves into the wash notch 50 thereby tripping the trip S switch 60 for signalling the microprocessor associated 9 4* with electronic module 200 (Figures 3 and 16) that the o,0. wash well is beneath the barrels 92. The trip switch 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.

I On the right hand side of the carriage 46 is notch 59 in which the roller 63 is shown. A sample plate trip switch 62 is thereby tripped to indicate that the carriage is at its maximum forward position. Trip switch 65 is mounted on the rear edge of carriage 46. Switch 65 closes when 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 when carriage 46 is in the maximum rearward 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 apparatius which will be explained in detail below. Lamp 212 provides a visual indication to the user of the S automatic pipetting apparatus 10 that the power is turned a on.

Figure 6 illustrates in a cross-sectional view taken along lines 6-6 of Figure 5 the means by which the o*0* carriage is translated with respect to tracks 34. The *c tracks 34 are supported upon base 12 by nmeans of supports 48. The carriage 46 includes slots 149 in its sides on which it slides on tracks 34.

Plate 16 as indicated above is disposed within a notch or valley 47 of the carriage 46, The carriage translation motor 40 is fixed to the base 12 and includes -21a shaft 42 which is supported by means of shaft supports 44. Pinions 38 secwued fto shaft 42 have their gears in engagement with racks 36 which 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 forward or rearward directions.

Figure 7 illustrates the pipette assembly 70 looking rearwardly along lines 7-7 of Figure 3. The mounting blocks 76 are shown secured to the base mounting blocks 78 by means of screws 79. The mounting blocks 76 carry vertical mounting po, ts 72 as illustrated in Figures 7, and 13 and in the top views of Figures 8, 11 and 14. The front plates 73 and back plate 86 are secured by means of scrows to mounting blocks 76.

t A mounting plate 80 is vertically slidably supported about the vertical mounting posts 72. Retainer bearings 74 provide sliding engagement between the posts 72 and vertical bearing blocks 75. The mounting plate 80 is fastened to extensions of bearing blocks 75 by means of screws 81. By reference to Figures 7 and 13, it is seen *that mounting plate 80 may be moved from its upward position as shown in Figure 7 to its lower position as shown in Figure 13 by its ittachmeant to bearing blocks °4 and their sliding engagement on ptsts 72.

A barrel bar 88 is secured to mou ing plate 80 Ly means of screws 89. Mounted on bf ,l bar 88 are a i -22plurality of pipette barrels 92 having their heads 193 secured within the barrel bar 88 in a manner to be described below. As illustrated in the partial cut away 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 extending below the bottom edge of the mounting plate which cooperate with 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 ips 93, when plungers 94 are driven downwardly wit '.arrels 92, to "kiss" or be slightly 44 S4.. applied either o ,he support medium or the blotting .s paper. The drcp~! on the lower tips 93 of the barrels o 92 are held beca a of their small size (as small as one micro liter) and surface tension forces of the barrel 0o 0 tips. When the tips are brought to a small distance within thE( upper surface of the support medium 20 or 4 blotting paper 30, the droplets are relieved of ths 4 *0 surface tension holding them to their barrels and are precisely applied to the blotting paper or to th support mediUtt.

0* Actuator guides 183 are secured to the mounting plate an, inctlude grooves in which an actuator plate 84 is inserted for sliding movement upwardly and downwardly with -23respect to the mounting plate 80. The actuator plate 84 has grooves in which a plunger bar 90 is inserted. The plungers 94 of the microsyringe barrels 92 are secured to the plunger bar 90 and extend within 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 downwardly with respect to the mounting plate 80, and through such action, the plunger bar 90 moves downwardly with respect to the barrel bar 88 causing plinlgers -4 to move downwardly within the barrels 92 thereby forcing any fluid within %uch barrels outwardly through the tips 93 of the barrels and forming a droplet at the tips of the barrels.

Position signals 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 4a 4 1 plungers 94 ;,ith respect to the mounting plate 80. The trip switch 106 mounted on the mounting plate cooperates with the lower stop 115 mounted on the mounting block 76 to provide a lower mounting plate position signal <O when the mounting plate 80 reaches its lower extent, ,n a r- 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 with the upper stop 114. The upper trip switch 105 when S tripped provides a signal to the microprocessor of c I -24electronics 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 with 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 and the trip switch 108 is tripped by wash cam 102 as the actuator plate 84 moves further upwardly. The down cam 104 trips trip switch 110 mounted on the right hahd side actuator guide 183 when the actuator plate 84 reaches its maximum downward travel where the plungers 94 are within the barrels 92.

Figure 9 is a cross-sectional view taken along lines 9-9 of Figure 7 and illustrates the mounting plate 80 and o .0 the actuator plate 84 both in their upward positions.

SMounting plate motor 116 has driven the mounting plate S° 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 now to Figure 10, the pipette assembly 70 is shown with the mounting plate 80 in the downward position but the plunger bar 90 and the plungers 94 are in their upward position with respect to the barrel bar 88. The S, barrels 92 are in a downward position in the wash well for aspirating 5 4p fluid, for example, from the wash well 26.

i -~111-rT~- Figure 11, a view of the pipette assembly looking downwardly along lines 11-11 from Figure 10, illustrates the arive mechanism by which the mounting plate 80 is moved up and down with respect to the base. A mounting plate actuator motor 116 is fixed to the mounting plate by means of a mounting screw 117. The output shaft of the motor 116 has a gear 119 fixed to it. Geat 119 is engaged with a left pinion gear 120L which is mechanically coupled to a right pinion gear 120R by means 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 10) 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 S° stop screws 95 of guide tips 97 are slightly above 9a 9 surfaces 49 of the plate 16 indicative that tips 93 are oat slightly above the top edge 49 of the plate so that droplets which form on the tips may "kiss" the surface 49 and apply sample fluid to the support medium or blotting paper.

-26- Figure 12 is a cross-sectional view of the pipette apparatus taken along lines 12-12 of Figure 10 and shows that the mounting plate actuator motor 116 has turned its pinion gear 120 so that the mounting plate 80 has been moved downwardly 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 where the barrels 92 of the pipettes have been lowered by the mounting plate 80 and where the fluid from the wash wells 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 with respect to the raised portion 22 by means of the motor 116 turning and causing the pinion 120 to translate upwardly and downwardly on fixed rack 122.

Turning now to Figure 13, the state of the pipette apparatus 70 is such that the actuator plate 84 has moved downwardly causing the plungers 94 to be inserted back into the barrels 92 thereby positively displacing any fluid which has been aspirated within the barrels either to an application space, a blotter, or to a waste well.

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 the wash cam 102 providing a means for signalling 0* o o oc o 0 0 0 66 a 6 090 4u 46 6 -27the position of the actuator plate 84 with respect to the mounting plate Figure 14 is a downward looking view along lines 14-14 o. Figure 13 and illustrates the drive mechanism by which the actuator plate 84 is translated vpwardly 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 naounting screw 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 is supported by means of shaft mountings 118 which also supports shaft 124 (see Figure 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 pinions 130L and 130R.

As the plunger bar actuator plate motor 126 is caused to turn in either the clockwise or the counterclockwise direction, the actuator plate 84 is caused to move 20 upwardly or downwardly with respect to the mounting plate 80. Figure 13 shows the slots 136 in the mounting plate 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 0000 0 @0 o 0 00 04 ofr 00 0J 0 0 *00 0d~ 0 0L1 *0 i i 1 9 -28means 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 screws 141. The barrel lock bar 91 is similarly constructed as the barrel bar 88.

The barrel bar 88 provide a removably securing means by which the barrels 92 may be easily replaced due to wear or breakage.

Figure 15 is a cross-sectional view looking along lines 15-15 of Figure 13 and illustrates the mounting plate 80 in a downward position. The actuator plate 84 has been translated downwardly where the plunger bar 90 is adjacent the barrel bar 88. Figure 15 illustrates the actuator plate racks 132 extending through slots of the mounting plate 80 and their engagement with pinion gear 130 which has been turned by means of the actuator plate motor 126. The barrel 92 is now in a rinse Nell 28, for example. The plunger 94 has been forced down by means of the actuator plate 84 moving the plunger bar 90 to its lowermost position. Of course, the raised portion of the plate 22 has moved longitudinally with respect to the pipette assembly between the views of Figures 12 and 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.

HD68P01V07 manufactured by the Hitachi Corporation. The

I

I I *0 4 4 44 040 044 4 40 4* o0I t 04 0 4401 4440' a-i 1 i r -29circuit 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 software by which the entire automatic operation is controlled and wil 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 as circuits UDN-2952B manufactured by the SPRAGUE Corporation. These motor driver circuits are used to control the speed of the motor and its direction of rotation. Also provided in conjunction with 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 position 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 with respect to the pipette assembly.

The position detector circuits 209 represent the t circuitry associated with the lower trip switch 106 and the upper trip switch 105 which signal the upward or downward limits of travel of the mounting plate 80 with respect to the base.

The position detector circuits 210 represent the circuitry associated with trip switches 108 and 110 which indicate the relative position of the actuator plate 84 with respect to the mounting plate 80. The signals associated with each of those position detector circuits are represented as being carried by a bundle of electrical leads 215 to the input interface circuitry 201F of electronic module 200.

The alarm circuit 211 is provided for the apparatus, for example, such as a sounding device EAF14R06C manufactured by Panasonic. Such circuit is activated 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 t illustrated in Figure 5 to signal the user that the power is on to the apparatus. 'he interlock tircuit 213 represents the circuitry with sample plate interlock trip o 20 switch 65 which indicates the presence or absence of the sample plate on the carriage. Command circuit 214 represents a push button switch used to start or abort the sn automatic pipetting application process.

In operation, the central processing unit 201A receives the sequence of events instructions from the I programs stored in the read only memory 201B. The central processing unit 201A then receives positional information -31concerning 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 microprocessor 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 which may be the push button 214 illustrated in Figure Figure 17 illustrates in flow chart form the operations of the CPU 201A under program control. The CPU 201A determines the validity of a cmmnand to start the processing by means of reading and decoding the binary coded data present at the input interface 201F which receives a signal from the interlock circuit 213. This operation insures that the plate 16 is fully inserted into the carriage.

Sa.. 20 The CPU 201A causes the motors 40, 116 or 126 to turn in the required direction be means of writing the appropriate binary coded data to the output interface circuit 201E which is connected to the motor drive circuits 202, 203 and 204. The microprocessor CPU 201A e then causes the mechanism movement to stop precisely when the required location is reached by writing the appropriate binary coded data to the output interface -32circuit 201E which is connected to the motor drive circuits 202, 203, 204 to disable the drive and then writing the appropriate binary coded data to the output interface 201E which 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 microprocessor CPU 201A to determine electrical 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 within the prescribed length of time, the drive command is aborted and the alarm is activated by means of the microprocessor CPU 201A writing the appropriate binary 20 coded data to the output interface connected to the alarm Scircuit 211. The timer 201D is also used to determine the repetition rate of the alarm thereby allowing the microprocessor circuit CPU 201A to e'Icript and communicate S. to the operator the nature of the failure.

As shown in Figure 17, once the automatic pipetting apparatus of the invention is running, a wide variiety of different applications may be achieved. The sequence of i j-- 1 -33operations shown in Figure 17 is preferred 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. ThBn the applicator tips are dried by lowering them to a blotter pad as illustrated in Figure Next, the barrels are moved to their upward position with the plungers in their downward position, the carriage is moved rearward and the barrels are lowered into the sample chambers 24. The plungers are raised thereby drawing 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 er 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 lowered such that distilled water is again drawn into the barrels and then dispensed into the 4, 20 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 flow Schart of Figure 17 branches to the dilute routine. A preferred routine for diluting the samples is to draw an additional four micro liters of fluid from the wash well.

This action results in each of the ba-Zels being filled Wl#4 -34with four micro liters of diluting fluid water) 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 row of the plate of Figure 2B. This process may be repeated a desired number of times to effect mixing of the sample with the dilution fluid (water). 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 where 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 wells. A predetermined Samount of wash liquid may then be applied to the dilution wells. After mixing the combination of the wash liquid °o0 20 and liquid sample of the dilution wells, (for example by the mixing technique described above), a sma.L amount of <^liquid samples is aspirated from the dilution wells and applied to the support medium.

The appendix to this specification includes a source listing of the computer program written in HD68P01V07 Hitachi Assembly language which is stored in the read ohly memory 201B so as to automatically ontrol the pipetting, cleansing, blotting, diluting (at the operator's option) and other functions described above.

Various modifications and alterations in the described structures will be apparent to those skillid i' the art of the foregoing description which 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.

o I 0 44 q, ,INTERNAL REGISTER MAP** 4 6 INPUT/OUTPUT PORTS* 7 Ct# ~4 0 0 o 000 S 00 o o o 0 #~a

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S t S6 0000 000i 0002 Pl.DDP E')U $0000 P2LDDP EQU $00051 Por~t I Dpta Dir, P e( Port Data Dir. R ea o Fort I. Dat egist er PORT t EQU $0002 'Pi t 0 p i t I S~i t2= Bi t83= B i t 4 t 5 Bi t6= Pi it7 (F (PI P 12 (P I q) (P14) (P15) (P16) (P17) Carrie Tbr-ate Not used Tube Tbrake Not used Ne-i'dle Thra'(e Needle Motor enable Tube Motor enable Carriaae Motor ena~ble $00024 Port 2 Data Regist Not Beeper QLutOut Not used~ Not use,4 Not ut-ed 003 POP.T2 EQU 27 28 so. ~c Bit0= Bitl= Bit2= Vit3= Bit4= (P20; (P21) (P22) (P23) (P24) 0004 0005 0006 P@DDR E,-1U $0004 PADDR EQU 00 05, TPICS3 EQU t$00 hPort Data Dir, R ea Port 4 Data Dir. F ag Port S Data Regist

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PlT2= (P30) (psi1) (P32) 1* J ITS= (P33) (P3'3) *BITG= (P86~) DIRECTION i'T FORCR TR ]~RECIN FPCRC MOTOR DIRECTION PIT FOR TUBE MOTOR DIRECTION BXT! FOR TUBES MOTOR DIVECTION BIT7 F 0R NEEDLES NiOT

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DIRECTION BIT FOR NEEDLES MIOT NOT USED or 11

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Bit7= (P37) NOT USED SCI= Not u xsed SC2= Not used PORT4 EQU $S0007 0007 Port 4 Data Pegist pi to= B i tI B i t2= ~i t3= B~k 1 4 Bi t5= PBitE'= Pi t7= (P40) (P41) P42) (P.43) (F44) (P4!5) (P46) (P47) Home aosition for carrier Home oosition Pcr tubheE Home taosition for needles Carri er position Tubes Dosition Needle position Not used No t use d **t a 6* *TIMER PEGISTEPS I 0003 "'CSREG EQU BITO= (OLVL) &.0008 Timer Control Stat us Reg.

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Input Captt4re Req Inpu~t Cap~ture Reg I CRMSB EQU I CVLSB EQU S 00 CD t00QE F ORT IRG CONTROL REGISTER

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88 89 91 92 92 94 96 97 99 99 C It 100 '1 102 t 0 103 t1iii: 10>4 105 107 108 109 110 I 111 1 2 t 113 I i 114 115 1 16 117 118 119 120 121 122 124 125 126 127 128 129 130 31 B BitO= a Bi;~= *Bi t2= B Bit3= PORT3 Latch Enable Biy4= OSS (Output Strobe Sel.) BitS= Bit6 IS.3 IRQl Enable Bit7= IS3 F)aa Bit SERIAL PORT REGISTERS 0010 RMCREG EQU $0010 Rate Mode Control Reg Bi tO= Bit I= Bi t2= Bi t3= Bi t4= BitS= Bi t6= Bi t7= (SSO) Baud Rate Select (661) Baud Rate Select (CCO) Format ClK Source (CCI) Format 01k Source 0011 TRCSFG EQU $0011 Xmit Recieve Cortr ol k Status Bi tO= Bit1= Bit2= Bi t3= Bit4= Bi t5= Bi t6= Bi t7= (WU) Wake-uo (TE) Xmit Enable (TIE) Xmit IPQ2 Enable (RE) Recv Enable (RIE) Recv IRQ"4 Ebable (TDRE) Xmit Data Reg Em~ty Flag, (ORDE) Overrur'Framiria Error (RDRF) Recieve Data Reg Full 0012 0013 RCVREG EQU 90012 XMTREG EQU $00,13 Receive Data Regis ter k Transmit Data Re,i ster *RAM* NE*P**OM*CONTROLREGISTER* RAM AND EPROII CO)NTROL REGISTERS 0014 RAMROM EQU $0014 Ram Eprom Control Pea 132 133 134 135 136 BitO= (PLC) Programing Latch Control Bitl= (PPC) Programing Power Control Bit2= Bit3=

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137 138 139 140 141 142 143 144 145 146 147 148 149 150 151 152 153 154 155 156 157 158 159 160 1 S.1 Bit4u BitG= (RAME) Enable Ram Addressing Bit7= (STBY PWR) Standby Power Status Flag $009i,-$001F RESERVED STANDBY RAM ($0080-00BF) TEMPORARY RAM ($00P 1 0-$OFF) 0080 0040 0008 0002 0004 0004 001 0 0017 IRQMSY EQU $80

TUBEENB

TUBEMOTP

TUBESW1

TUMJETBRK

TUBEMOTN

TUBESW2

TUBEMSK

EQU

EQU

EQU

EQU

EQU

EQU

EQU

$40 $08 $02 S04 $04 $10 $F3 162 163 4 '1 164 165 166 167 1E 169 0080 0002 0001 0001 0001 0008

OOFC

CRGCENBI

.CRGMOTP

C PGOSWI

CRGTBRK

CRGMOTN

CRGSW2

CRGMSK

EQU

EOU

EQU

EQU

EQU

EQU

EOU

$80 $02 $01 $01 $01 $08

$FC

DEBOUNCE MASY STAR T SWTCH *TUBE DRIVE ENABLE *TUBE MOTOR DRIVE P0 SITI VE.

*TUBE HOME SWITCH.

*TUBE TRIAC BRAIE *TUBE MOTOR DRIVE NE

CATIVE,

*TUBE DOWN POSITION

SWITCH,

*MASX<S OFF THE TUBE DIRECTION BITS TO A 0.

*CARRIAGE DRIVE ENAB *CARRIAGE MOTOR DRJV E POSTIVE.

*CARRIAGE HOME POSIT ION SWITCHt *CARRIACE TRIAC BRAK

E

*CARRIAGE MOTOR DRIV E NEGATIVE, *CARPIAGE LOCATION S

WITCH.

*MASI'S OFF THE ORG D IRECTION BIT TO A 0.

*MASK OFF THE NEEDLE DIRECTION BITS TO A 0 *NEEDLE MOTOR DRIVE

POSTIVE.

*NEEDLE HOME POSITIO 170 171 172 173 0020 0020 0004 NDLENB EQU $20

NDLMOTP

NDLSWI

EQU

EQU

$20 $04 i4 1t Le :1: 3:-e- 174 175 176 177 178 179 *#.180 *184 ***182 187

IS,

184 190 196 192 194 190 191 192 198 199 290 216 200 203 204 0010 0010 0020

OOCF

0003 0040 0001 0002 0003 0004 0005 0006 0001 0002 0001 0002 0003 0004 0004 00OFF 0015 0QE 0 0002 00 OF 5000 02FF 04FF 0 8FF

OFFF

I FFF

NDLTBRI(

NDLMOTN

NDLSW2

NDLMSK

EQU

EQU

EQU

EQU

CLEANYET EQU

TRAY

WASH

WASTE

BLOT

SAMP WELL

DULUTER

PLATE

NDLIUL

NDL5UL

ERROR!

ERROR2 ERROR3

ERRORA

ETO I $3TACK (4L BRAK

ALLENB

BRAKEPRT

SWBOUNCE

JOGTIME

TONE 1 TONE2

TONES

TONE4

EQ'J

EOU

EOU

EQU

EQU

EQU

EQU

EQU

EQU

EQU

EQU

EOU

EQU

EQU

EQU

EQU

EQU

EQU

EQU

EQU

EQU

EQU

$10 $20

$CF

103 $40 $02 $03 $04 $05 $06 $01 $02 $01 $02 $03 $04

$EO

FORT I

$OF

$5000 $O2FF SO4FF

$OSFF

$OFFF

N SWITCH.

*NEEDLE TRIAC BRAKE *NEEDLE MOTOR DRIVE

NEGATIVE.

*NEEDLE LOCATION SW!

TCH,

*MASK( OFF THE NDL DI RECTION BIT TO A 0.

*HOW MANY TIMES T1O C YCLE THROUGH THE WAS H CYCLE.

*TO MAKE SURE THE TR AY IS ALL THE IN.

*CARRIAGE LOCATION 0

NE.

*CARRIAGE LOCATION T

WO.

*CARRIAGE LOCATION T

HREE.

*CARRIAGE LOCATION F

OUR.

*CARRIAGE LOCATION F

IVE.

*CARRIAGE LOCATION S

IX,.

*NEEDLE LOCATION ONE *NEEDLE LOCATION TWO *TUBES NOT AT HOME P

OSITION.

*MOTOR NOT D RIVING.

*TRAY IS'NOT ALL THE WAY IN. 1 *TIMER OVERFLOW FLAG IN THE TCSREC, *TOP OF RAM.

*ALL MOTOR ENABLE *WAITING FOR TU)BE SW ITCH TO STOP BOUNCIN *TUBE HOME POS. ERRO R TONE.

*MOTOR NOT REACHING LIMIT SWITCH TONE *TRAY NOT IN TONE.

*SWITCH NOT CLOSED E

PROR.

*UNKNOWN ERROR, TONE5 EQU $1FFF 1.1 l -1 rj~F 7 r.

.4 o .r 94 94 9 205 206 207 208 209 210 211 212 213 21<4 215 216 217 218 219 220 221 S22 224 225 228 229 230 231 1FFF TONE 0007 0006

OOFF

00AF 0010 0080 0080 0081 0083 0084

SWPRT

MOTORPRT

OFFSW

SOAKTIME

BLOTIME

WASHTIMS

TONES

WATCHDOG

ENABLE

EQU

EQU

EQU

EQU

EQU

ORG

RMB

RMB

RMB

RMB

PORT4

PORTS

$OOFF

$OOAF

$0010 2 1 1 EQU $iFFF *READY FOR OPERATION

TONE.

*SWITCH PORT, PORT4 *MOTOR PORT PORTS *PAUSE TIME.

MOTOR CONTROL BLOC K ENAPl MOTOR CONTROL BLOC K DIRECTfON MOTOR CONTROL BLOC K SWITCH MOTOR CONTROL BLOC K TRIAC BRAKE MOTOR CONTROL DIRE CTION BIT MASK Mode FtaQ 0085 DIRECT RMB I 0088 SWITC" RMB 1 0087 TDRAYKE RMB 1 0088 MASK 0089 00SA )8B 008C

CRGPOS

NDLPOS

STATUS

COUNT

RMB I RMB 1 RMB 1 RMB I RMB 1 MAC START LDAA ENABLE

COMA

ANDA BRAKEPRT STAA BRAEPRT LDAA MASK ANDA MOTORPRT ORAA DIRECT DISABLE DRIVE ZERO DIRECTION BIT

S

GET REQUIRED DIREC

TION

ENABLE

233 234 285 238 237 245' 238 240 241 242 243 244 245 246 247 24.

249 250 490 F OOSD

STAA

LDAA

ORAA

STAA

MEN

MAC

LDAA

COMA

ANDA

ORAA

STAA

LDAA

ANDA

STAA

LDAA

ORAA

STAA

MEN

MOTORPRT

ENABLE

BRAKEPRT

BRAKEPRT

STOP

ENA ELE

BRAEPRT

TBRAKE

BRAK EPRT

MASK

MOTORPRT

MOTORPRT

ENABLE

BRAKEPRT

BRAKEPRT

DISABLE DPVE TRIAC BRAKE BRAKE AND DISABLE DRIVE FAST STQP DRIVE FAST STOP 008D 4k' ~ur~ 7-35gt tr I it it 0 I I *0 0 #6*00 251, 252 254 255 256 257 252 259 260 261 262 263 264 265 266 267 258 269 270 2R 8 14 I it 20 F800 F800 P801 F804 F807 F809

FSOB

FSOC

F8OE F8OF FS11 F812 F814 008B 008C 02 01

ORG

INT' SE]

CLP

CL

LDAA

STAA

SF800

STATUS

COUNT

#$02 P2DDR 4F 97 08 43 C6 EO 53 97 00 D7 02

CLRPA

STAA

COMA

LDAB

COME

STAA

STAB

TCSREG

#ALLENB

P DI)R PORT I ALL OUTPUT BRAKEPRT SET ALL TRIAC BRAK E,DISABLE DRIVE P3DDR PORT 3 ALL OUTPUT *MAKES P21 AN OUTPUT 271 272 273 274 275 276 277 278 279 280 281 282 283 284 285 44 F816 47 F818 6R FSIB 8 F81E 11 820 15 F822 18 F824 22 F822' 26 F8231 2R F87,A 11 *FF92B 20 *F82E 29 *F831 38 *F834 44 F837 46 F83A 51 F83C 60 F83F 2R F840 4 F842 8 F843 04 7FFF 0006

EO

02 02

LDAP

ORAB

STAB

STAA

LDX

H1 CLR 8E OOFF 09 26 Fl

GE

BD F9DA BD F9F4 BD FA23 BD FB26 7F 0080 86 80 B7 008B 3E C6 EO 53 D7 02 (F8 iB)

LIDS

DEX

BNE

CLI

JSR

JSR

JSR

JSR

CLR

LDAA

STAA

WAI

EXEC LDAB "0Mp.

#$7FFF

MOTORPRT

#ALLENE:

IWAKEPRT

BRAKEPRT

#STACK

!I

TUBEHOME

CRGHOME

NDLHOME

BEEPER

WASHTIMS

STATUS

#ALLENB

ENABLE

AST STOP DRIVE.FOR F MOTOR DRIVES FAST

STOP

Mode READY STAB BRAKEPRT *SETS THE TRIAC BRAK TS AND DISENABLES T MIE MOTORS.

CLR MOTORPRT *MtjTOR DRIVERS FAST

STOP-

286 14 F845 7F 0006 287 16 288 19 289 23 290 291 292 293 294 295 296 F848 F84A F84C F84E F851 F853 F855 F857 F858

LDAI

GR41B

STAB

OALLENB

BRAKEPRT

BRAKEPRT

#QF'FSW

APRT

#IROMSK

17 *ENABLES MOTOR DRIVE RS FOR FAST STOP.

CE OOFF 96 07 84 80 27 F7 09 26 F7 17 LDX 14 LDAA

ANDA

BEQ

DEX

3NE (F84E) (F851) i 14--

CEBBO)

297 BR FB5A 7D 008B 298 10 FB5D 26 01 299 20 F85F B 300 24 FB6O 28 03 3601 3R FS62 7E F800 TST ST AT() ENE !'2

RTI

(F865) 12 EM] If Not 0 Not INT T'L Mod c if READY Mode so co RUN If RUN Mode so o TWIT JM",p INIT

A,

*AW

tA

A

IAA 4A

A

4 ~AA Aft t~

A'

i A AAA 302 303? 304 305 30 e 307 !308 309 310 3911 312 313 314 315 316 317 318 319 321-0 A I ~I 321 323 324 325 326 327 328 8A 29 1430 331 332 335 337 338 339 340 341 342 343 34a, 2R F865 7 F867 9 FB6A 12 F86.B 14 F86E 23 *F8 7 0 32 *F873 34 FB76 43 *FB8e.

52 *FS7B 54 FS7E 63 *Fe80 72 *FBEZI at *F886 90 *F889 92 F88C 101 *F88E 110 *F891 119 *F894 128 *F897 1.30 EB9A 13 9 *F69C 1.48 *F89F 150 F8A2 159 *FBA4 1GB *FSA7 t70 FBAA 179 *FBAC 188 *FSAF 197 *FBB2 206 *FBB5 215 *F8BB 217 F8BB 226 *FSBD 235 *FSC0 237 F8C3 246 *F80C5 255 *FBC8 258 F8CI3 260 FBCP 264 FS%"F 2R~ F8D.

It *F8D3 20 *F8D6 ?9 *FBD9 F9DA 04

FACER

01 FAF7 F9DA 03 FA CS F %23 F9E7 FC44 F9DA 04

FACS

F9E7 01 17AF7

FSDA

4C 05 'FAG S PA23 VS E7 B6 7 F B7 00813 0OE BE 00FF C 6 01 ED FACS PD F9E7 CE 6 01 ED FAF7 ED F9DA C 6 02 ED FACS B D FA23 BD F9E7 ED F9DA 06C 03 BD FACS ED F9E7 ED FC44 1B LDAA

STAA

CL I

LDS

LDAB

JSP

J ER J SR

LDAE

Jsp dSP dSR i -p

!,DAB

ISR

JSP

Jsp I SR

JSP

LDAB

LDAB

lisp J SR

ISP

i S R

LDAB

~JS5

LDAA

ANDA

BE

LDAB

I SR J1SR

J$R

#~STAC K

#WASH

C P GLO0C

TUBEDOWN

NDLLOC

TUBEHOME

#CWA STE

CFQLOC

NDLHOME

TUB EDO'WN

TUBEHOME

hVLOT CR CLOG

TUBEDOWN

BLOTTER

TUB EHOME 4~SAMP WELL C PCLOC

TUBEDOWN

#NDLIUL

N DL1.LO C

TUBFHOME

#CBLOT

CRGLQC

NDLHOME

TUBEDOWN

BLOTTER

TUBEHOMIS

ISAMPWELL

C.RGIOC

TUBEDOWN

ICWDLIUL

NDLLQC

TUBEHOME

MOTOPFT

110

ODULL)TER

CRGLOC

NDLHOME'

TUBEDW6C? *THE TIMlE THE TUBES WILIL STAY DOWN4 #$7F ST ATUS Flag RUN mode (59 10D) '4

<S

j S I k t a r4~ a..

a. a.

a.

347 38 *F8DC 348 47 *FSDF 349 49 FSE2 350 58 *F8E4 351 67 *F8E7 352 69 FSEA 353 78 *FSEC 354 87 *FSEF 355 89 FSF2 356 98 *F8F4 857 107 *FSF7 358 116 *FSFA 359 118 F8FD 360 127 *FBFF 361 136 *F90'! 362 138 F90~5 363 147 *F907 364 156 *F90A 365 158 F90D 366 167 *F90F 3(67 176 *F912 '368 178 4=915 369 187 *F917 370 196 *F9tA 3(71 2R F91D 372 11 *F91F 373 20 *F9247 374 29 *F925 375 98 *F928 376 47 *F92B 377 49 F92E 378 58 *F930 379 67 *F938 380 76 *F936 S8l 85 *F939 382 2R F93~C 8So3 11, i '3E 384 20 *S4 1 382-5 F 9 44 3o86 -;81 '*F946 387 40 *F949 388 42 F94C ,389 51 *F9 AE 390 60 *F951 391 69 *F954 392 78 *F957 393 82 F95A 394 84 F95D 395 88 F95F 396 2R F961 397 6 F963 398 2R F965 3899 6 F967 400 2R F969 ED FC44 ED F9DA 06 01 ED FACS ED F9E7 CE6 02 ED FAF7 ED F9DA~ C06 05 E.D FACE ED FA2Z4 ED F9E7 C6 02 ED FAF7 BD FA23 C6 02 ED FAF7 RD FA23 CE 02 ED FAF7 ED FA2a c 6 01 ED FAF7 I':D F9DA\ 06 06 ED F AC S BD FA23 ED F9E7 ED F AB ED F9DA C 6 0 I= BD FACE ED F9E7 ED FC44 ED F9DA C6 01 BD FAiCE ED F9E7 C6 02 ED FAF7 ED F9DA C6 02 PD FACE ED FA23 ED F9E7 ED F9DA B6 0080 '81 00 27 08 at 01 27 -"F 81 02 27r 12, C6 0-4 Lisp isp

LDAB

Li El I DAB LD PB LiSR

LDAE:

isp

I-DAB

i -op LiSR

LDAB

Li SP isp isp

LDAB

i s i $p i SFP LiSF Li SIR

LDAB

"SRp isp

LDAB

LiSP Li SRp LiSR

LDAA

OMPA

BEG

CMPA

BEG

OMpA

BEG

LIDAB

BLOTTER

TUBEHOME

#WA~SH

CRGLOC

TUBE DO WN SNDL5U L

NDLLOC

TUREHOTIE

#DULUTEP

CRGLOC

NDLHOTIE

TUBED OWN NDLS UL NDLL 00

NDLHOME

NDLSU L

NDLLOC

NDLHOfIE NDLLO C

NDLHOME

4NDLIUL

NDLLOC

TUBEHOME

#PLATE

C ROL 00 NDLHOfIE

TUBEDOWN

SOAY'

TUBEH OME MtBOT CFGCLO C

TUBEDOWN

BLOTTER

TUBEHOP,,

OtWASH

CPC.LOC

TIJEEDOWN

4tNDL.SU L

NDL(CC

TUBEHCiIE OtWASTE CROCLO C

NDLHOME

TUBEDOWN

TUBEHOME

WASNT IMS #$00 1~ (F969) (F994) F97 B

A

a., 0-4 rr 401 11 *F96B 3D FAC8 402 14 F96E CE 5000 JSR CRGLOC LDX *JOCTIME F971 F973 *F976 F979 F97B *F97D F980 F982 F985 r* I 4 I (I 412 413 414 415 4li 419 420 421 422 422 422 422 422 422 422 30 *F988 9R*F982 18 *F9E 27 *F991 ,R F994 F997 F999 16 F99C 3r F99E 9R*F9AI F9A4 13 F9A4 15 F9A7 18 F9A8 22 F9AA 26 F9AC 29 F9AF 86 B7

BD

20 C6

ED

86

CE

BD

BP

BD

BD

7C 86 B1 26 7E

PD

B6 43 94 97 B6 94 02 0085 F9A1 10 03

FACS

01 0085 5000 F9A1 F9E7 FC44 F9DA 0080 03 0080 9E F800 FC53 0084 02 02 0088 06 (F98B) (F93C)

LDAA

STAA

JSR

BRA

is LDAB

JSR

t,DAA

STAA

LDX

J SR !9 JSR

JSR

JSR

16 INC

LDAA

CMPA

BNE

JiP JOG JSR

START

LDAA

COMA

ANDA

STAA

LDAA

ANDA

O4AA

STAA

LDAA

ORAA

STAR

Ii DEX

BNE

STOP

LDAA

COMA

ANDA

ORAA

9TAA

LDAA

ANDA

STAA

LDAA

ORAA

STAA

RTS

TOBEHOME LDAB

LDX

#CRGMOTP

DIRECT

JOG

!9

ILOT

CPGLOC

#CRGMOTN

DIRECT

#JOGTIME

JOt

TUBEDOWN

BLOTTEP

TUBEHOME

WASHTIMS

*CLEANYET

WASHTIMS

II

INIT

PRAK OFF

ENABLE

BRAKEPRT

BRAKEPRT

MASK

MOTORPRT

DIRECT

MOTOPRT

ENPA8LE

BRAKEPRT

BRAKEPRT

Ii

ENABLE

BRAKEPRT

TBRAKE

BRAKEPRT

MASK

MOTORPRT

MCTOPPRT

ENAdLE

BRAKEPRT

BRAKEPRT

*HOW LONG TO LEAVE T HE MOTORS ON.

*HOW LONG TO LEAVE T HE MOTORS ON DISABLE DRIVE ZERO DIRECTION BIT

S

GET REQUIRED DIPEC

TION

ENABLE

422 33 *F9P1 BA 0085 422 422 422 42.

42- 424 425 425 425 425 425 S0 425 425 425 426 427 428 F91P4 F9B6 F9B9 F9B F9 DD F9PE F9CO F9CO F9C3 F9C4 F9C6 F9C9 F9CB F9CE F9DQ F9f2 F9D7 F9D9 F9DA F9DC $7 06 86 0084 9A 02 97 02 09 26 FD P.S 0084 43 94 02 BA 0087 91 02 Bs oO88 94: OE 97 QS B6 00$4 BA 02 97 02 39 CS 01 CE F9E2

(FPD)

DISABLE DRIVE TRIAC BRAKE PRAKE AND DISABLE TR7VE FAST STOP DRIVE FAST SiOP $S 01 oil .w -w

I

1 *4* 4 44 It I 1 *4 4 I *41

I

I 11444 K I

II

I I

I

429 43 0 431 4:32 433 434 48 6 4'as 4:39 440 441 442 443 44-4 445 446 447 448 4413 4.50 4,51 452 455 4!56 457 45.0 459 460 461 462 463 464 4 r,5 466 467 468 469 470 471 472 473 474 475 476 478 470 480 482 6 F91)F F9E2 F9E? F9E4 F9E5 F9E6 2P F9E7 5 F9E9 8 F9EC F9FO F9F3O 6P F9F4 15 *F9F7 1S F9FA 20 F9FD, FA03 FA04

FAOS

C. .06 9R*FAQ7 12 FAQA 15 FAOD FA 10

FAII

F A 12

FAIS

FA14 1 2 FAIS 12 FA I1S FAtE

FAIF

FA20 FA21 F A2 6P FA23 5426 11 F A2,0, 14 FA2B FA2E FA21F FA30 F AS I FA82 3R A? 3 6 FA36 F A S9 FAq-A

FASB

7 E FB7A 40 08 02 0.4 F CE 01 CE F9EF 7E FB77 40 04 to 04

FS

7F 00w09 I D FA3E CE FA02 CC 01 7E FB77 so 02 01 01

FQ

BD FASE CE FA10 7E FB77 so 02 08 01

FC

BD FASE CF, FAtE 7E Fly'77 s0 012 08 01 7F 0084 CE FAZE 20 20 Q4 10 0,F C E FA39 7E FB7A 20 la 20 imp I FCB

FCB

FCB

FOB

TUB'EDOWN LDAB

LDX

imp !I FCB

FQB

FCB,

FCB

CRGHOfIE OLP J SR

LD)X

LDAB

im it FCB

FOB,

FOB

FOB

FOB

CROBACI( JEP

LD)X

it FCB

FOB

FOB

FCB

FOB

CRGFRONT JSP

LDX

it

FOB

FOB

FB

FOB

FOB

NDLHOIE CLP

LDAP

LDX

FCB

FOB

FOB

NDL1UL5 LUX !I FCB FCtB

FOB

DRIVE!9

TUBEENB

TUBEMOTP

TUBESWI

TUBETBJ(.

TUBE!ISK

DRIVE

TUBEENB

TUBEMOTN

TUPESW2 TUBE TB RY

TIJBEMS(

C PC.POS C HEC KTUB

DRIVE

C RGENP? CRcSWI

OPCTBRK

CRGMSY

O HEC KTUB #11

DRIVE

CFGENB

ORGMOTN

CRGSW2 CRGC1SK C HEC K TUB #11

DRIVE

CRGENB

CRO.MOTP

ORGSW2,

ORGTBRK

NDLPOS

#$0 NDLEr2*T

NDLMOTP

NDLS4I

NDLTBRIK

NDLMS<

01 D I VE IS

NDLENB

NDLmQTN NDLsw2 I I I I

II

4 I I II 0~ 1*11* I I *1 0 r.

-x -351- 483 484 485 486 487 488 489 490 491 492 493 494 495 496 497 498 499 500 501 502 .902 502 502 502 502 502 502 502 502 502 502 503 504 505 506 507 i, 508 509 510 511 t:12 ~51A 51$ 519 5320 521 522 523 524 525 FA3C FA3D 2R FA3E 3R FA40 4R FA43 8 FA44 2R FA46 6 FA47 2R FA49 5 FA4B 9 FA4D 2R F A4F 5 FA51 10 FA54 3R FA55 6 FA57 12 FA59 16 FA5C 26 FASE

FASF

4R FA5F 6 FA62 9 FA63 13 FA65 17 FA68 21 FASA 24 FA6D 28 FA6F 32 FA71 35 FA74 39 FA76 42 FA78 44 FAZA 48 FA7C 50 V'A7E 52 FA7F 55 FA~si 2R FAS4 5 FAS6 9 FASS 14 FA8A 9P*FASB 18 *FA8C 2 r *FA91 29 FA94 T, 8 4A96 47 FA99 6R FA9A 9R*FA9D 3R FAAO 4R FAA3 8 FAA4 4R FAAS 6 FAA9 10

CF

86 OF CE 03FF 09 26 Fl 4A 26 F7 86 02 94 07 27 05 86 01l 7E FB49 39 96 08 96 09 7C 0083 27 01 3B 56 0084 43 94 02 BA 0087 97 02 B6 0088 94 06 97 06 56 0084 9A\ 02 l7 02 16 08 84 FB 97 08

OE

7E FB49 86 40 94 07 26 01 DD FA23 DD F9SDA BD FC44 CC 02 BD FAF7

SE

FF 0081 BD FB29 CE 0000 08 26 FD B6 O0PC 4C

FCB

FCB

CHECKTUB LDAA 12 LDX !I DEX (FA43) BNE

DECA

(FA40) BNE

LDAA

ANDA

(FA54) SEQ

LDAA

Jmp !3 RTS UPDATIME LDAA

LDAA

INC

(FA5F) SEQ

RTI

I!

STOP

+11 LDAA

COMA

ANDA

OPAA

STAA

LDAA

ANDA

STAA

14DAA

ORAA

STAA

LDAA

ANDA

STAA

CLI

LDAA

iMP TRAYIN LDAA

ANDA

(FABB) BNE

RTS

I8R

SR

JSR

LDAB

J SR

WAI

FLASH STX 12 JSR 13 LDX !I INX (FAA3) BNE

LDAA

INCA

NDLTBRK

NDLMSY

#SWBOUNCE

#$03FF

II

12 #TUBESW1

SWPRT

13 #ERROR 1

BEEPERRO

TCSREG

TCSREG+1

WATCHDOG

!I

ENABLE

BRAKEPRT

TBRAKE

BRAKEPRT

MASK

MOTORPRT

MOT0RPRT

ENABLE

BRAKEPRT

BRAI(EPRT

TCSREG

TCSREG

#ERROR2

BEEPERRO

#TRAY

SWPRT

!iI

NDLHOME

TUBEHOME

BLOTTER

#NDLSUL

NDLLOC

TONES

BEEPERI1 #s$0000 11

COUNT

*CLEARS IPQ DISABLE DRIVE TRIAC BRAKE BRAKE AND DISABLE DRIVE FAST STOP DRIVE FAST STOP 4-"

I

I

It I Itr t t t

II

I I

I

526 527 528 529 ~3 0 53 2 53? 53-4 535 537 53S 539 540 541 542 543 544 545 546 547 546~ 549 550 551 552 5 3 554 555 556 557 558 560 561 562 63 564 56G5- 566 .567 568 569 570 571 572 573 574 575 576 577 578 579 I1I FAAA 13Z FAAD 17 FAAF GR FABI I1I FABA 15 FAP7 3R FAD9 aR FABC 6 FABE 8 FACO 12 FAC2 4P FAC4 8 FAGS 13 FAC7 2R FACS 6 FAGS 10 FACC 2R FACE 7 FACF 9 FAD2 12 FAD4 16 FADS 2R FADS 5 FADA 9 FADO 2R FADE 3R FAEO 5R FAE9 7 FAE6 10 FAES 14 FAEA 2R FAEC 5 FAEE 9 FAFO 2R FAF2 3R FAF4 2A FAF7 S FPIFS 10 FAFB 2R FAFD 7 FAFE 9 FEO 1 12 FPO3 16 FEDS 2R FV07 5 FBO9 9 FBOB 2F FEOD 3R FPOF 5R FE 12 7 FB 15 10 FB17 14 FEIS 2R, MEi R7 OOSC 81 01 26 EF 7F 008C FE 0081 20 E4 CE QOAF 96 0S D6 09 84 20 27 FS 09 26 F5 39 17 FO 0089 24 15 50 B7 0089 86 01 9-4 07 27 08 86 08 94 07 26 02 CE 01 7'E FA15 B7 0089 86 01 94 07 27 02'- 86 08 94 07 26 02 CE 01 7E FA07 17 FO 0OQ8A 24 l b 50 B7 008A 86 04 27( 08 86 20 94 07 26 02 7E FA23 B7 QO8A 86 04 94 07 27 08 86 20

(FABC)

(FADC)

(FAE3)

(FAA')

(FA9D)

(FAEO)

(FAEO)

STAA

CMPA

ENE

CLF

LDX

EPA

SOAI' L-DX i tLDAA I DAB

ANDA

EEO

flEX EN E

PTS

CGLOC TP

SUB

EGG

NEGE

STAA

LDA A

ANDA

EQ

LDAA

ANDA

E NE

ADDE

12 ip i ISTAA

LDAA

ANDA

EEO

LDAA

A NDA

ENE

ADDP

13 'IMP NDLLQC I PA

NEG

STA A

LDAA

ANDA

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Claims (34)

1. Automatic pipetting apparatus comprising a base, sample plate means disposed on said base, said sample plate means having longitudinal and lateral dimensions, said plate means including a lateral 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 iquid isamples, a pipette frame including vertical support means for supporting said frame from said base laterally above said sample plate means, said pipette frame having a longitudinal position with respect to said longitudinal dimension of said sample plate means, logitudinal translation means for changing the longitudinal position between said pipette frame and said sample plate means, l a mounting plate carried by said pipette frame, vertical translation means for effecting relative vertical movement of said mounting plate and Ssaid sample plate m6ans, v o^ 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, 37 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 t. relative to said barrels, and *t 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 :1,1 apply a droplet of said liquid samples in each of said pipette barrels onto corresponding spaces of said microporous support medium when placed on said oo* lateral application space of said sample plate by precisely positioning said lower tips of said barrels *aat a small distance above said microporous support a 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 f tension forces or 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, I 38

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 whertein said pipette frame is translated longitudinally above said sample plate means by said longitudinal translation means,

4. The apparatus,, of. cl.aiq 1 wherein said sample t plate means includes a wash well adaittid to contain wash liquid and a waste well longitudinally spaced from each other and from said sample chambers, said programmed computer means genierating a further sequence of control signals to said lofgitudinal translation means, to said vertical translation means and to said plunger translation means before aspirating liquid from said sampleA 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 o waste well.

5. Automatic pipetting apparatus comprising a O, base, sample plate means disposed on said base, said sample plate means having longitudinal and lateral dimensions, said plate means including a lateral 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 microporouIs support medium, said 01 S /y P 4 39 sample chambers adapted to receive liquid samples, a pipette frame including vertical support means for supporting said frame from said base laterally above said sample plate means, said pipette frame having a longitudinal position with respect to said longitudinal dimension of said sample plate means, longitudinal translation means for changing the relative longitudinal position between said pipette frame and said sample plate means, a mounting plate carried by said pipette S" frame, I" t t 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 correspoiding 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 S*1 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 iiith respect to said sample plate meanst for generating mounting pt signa4s representative of the vw.trial oiBtoin o. i s mounting plate relative to said sprnoe plate means ana for generating representative of the td 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'miroporous support medium onto corresponding spaces of said microporous support medium when placed on said lateral application space of said sample plate, wherein said sample plate means includes a wash well adapted to contain wash liquid and a waste well longitudinally spaced from each other and from said sample chambers, .Ott said programmed computer means generating a further sequence of control signals to said a 84 4oo* longitudinal translation means, to said vertical translation means and to said plunger translation means before aspirating liquid from said sample Sichambers, o. to draw a second predetermined amount of *o vwash liquid from said wash well into respective S" pipette barrels, and to discharge said wash liquid into said waste well, and 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, seid wash well, said waste well and said lateral i /LT 41 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-a row 'of -individual' c* liquid dilution wells longitudinally spaced from said sample chambers, said dilution wells adapted to c( receive dilution liquid, said programmed computer means generating a t 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, ^I ,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 o, well 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, ,t PC It I I te ey 2 42 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 S, into said correstioding~ d'i'ilhtion wells 'operabiy 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 *t said programmed computer means generates a further sequence of control signals to said plunger translation means to alternately discharge said .sample liquid/wash liquid into said corresponding oB,, 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 I. 43 application space adapted to receive a microporous support medium and including a raised portion and a lower: portion, said row of indi.'",al 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.

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 weXl 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 4nd said lateral application space.

11. The sample plate of claim 10 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 and said blotting space being disposed on said lower portion. .ii- 4-a

12. The sample plate of claim 10 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.

13. The sample plate of claim 9 further *0 9 «4 t1 t 4 4 4 0 S i comprising a row of individual liquid dilution wells longitudinally spaced from said sample chambers.

14. 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 t ,of individual, liquid sample chambers and a lateral application space longitudinally separated from said 4 tt 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 frome 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 o said sample plate and including, a mounting plate having slidable guides diz osed about said posts, "e 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 r u I 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 S* indicative of plunger bar position, and programmed computer means responsive to said carriage position signals, to said mounting plate 0position 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 pipt.,e barrels, and to apply a droplet of 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 by precisely positioning said lower tips of said barrels at a small distance above /€flA L 46 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. 1,

15. The apparatus of claim 14 wherein said t ,t sample plate includes a Wasih well and a waste well longitudinally spaced from each other and from said Ssample 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, 1. to discharge said rinse liquid .nto said waste well. 4

16. Automatic pipetting apparatus comprising, S" a base, .o 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 -M 47 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 he;ads 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 o4* 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 48 vertically with respect to said basE ard 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 V, •signals for generating a sequence of control signals S, 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, and wherein said sample plate includes a wash wel! 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 ©3 ki 1 *i v 49 to discharge said rinse liquid into said waste well and wherein said sample plane 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 meains after discharging said rinse liquid into said.waste well, to blot the tips of sa'id barrels -on Sacid' blotting paper strip.

17. The apparatus of claim 16 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.

18. The apparatus of claim 16 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.

19. The apparatus of claim 14 wherein said translation and signalling means comprises, carriage translation means responsive to carriage translative signals for longitudinally translating said carriage and said sample plate and i;-i said track means beneath said pipette assembly for operably translating said plate to at least a sample chamber position whai 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 when said microporous support medium is beneath said microsyringe barrels, mounting plate assembly translating means responsive to up and down mounting plate assembly k translation signals for vertically translating said c 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 barrel into said fluid sample chambers when said plate is in said sample chamber position and operably allowing said barrels to be slightly abjve 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 a Sa mounting plate lower position signal when said M mounting plate is in said lower position, f 0 a plunger act~uator plate translating means responsive to up and down plunger actuator plate translation signals for vertically translating said plunger actuator pl.ate and said plunger bar between a lower position where said micro-plungers are fully within said corresponding microsyringe barrels toa sample upper position where said imicro-plungers are extended upwardly through sai~d barrels operably I w; I- drawing fluid into said barrels when said tips are Mersed 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 v° communication between said programmed digital computer and t said carriage translation means, said mounting plate assembll 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 t ~said plunger actuator plate signalling means, o

20. The apparatus of claim 19 wherein said a digital computer means includes a stored sequence of instruction for ad* oe 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 lild 52 plate assembly translation signal to said mounting plate assembly from said upper position to said lower position and removing :aid mounting plate assembly translation signal when said mounting plate lower position signal is received, said programmed digital computE~ means operably generating and applying said up plunger actuator plate translation signal to said plunger artuatur 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 programme- 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 p 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 carrigs3 position signal is received, said prograwmed digital computer means op-;ably generating and applying said down mounting plate assembly translation signal to said mounting plate assembly translating means thereby translating 0 rr -i I n; 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 qsign and removing said plunger actuator plate signal when said plunger bar sample S.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 f lateral application space of said plate. 4

21. The apparatus of claim 20 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 a4we 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 I 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 includes a washj.t. rins well pc iti and IIL_ t tI 41t1 t *T 4 *r b SW 4 -*4 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 instruction 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 aspirate into said barrels, said programmed cl -l 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 l- *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 S"r said mounting plate assembly from said upper position of*, 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 meanu. operably generating and applying said down plunger actuator plate translation signal to said plunger actuator plate translating means thereby translating said plunger form 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 fluLid in each of said barrels to be applied to said waste well, and said programmed digital computer means 56 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.

22. Automatic pipetting apparatus comprising, a base, track means disposed longitudinally on said base, n a carriage longitudinally movably disposed on said track means, t a sample plate removably disposed on said carriage, said sample plate including a lateral row of individual liquid sample chambers and a lateral,. \in VrQ :S Scev.o W\r 5.eiy V!,O A cSCLc%&,,ta k:cs'I-; So& application spacelateral application space adapted to receive a microporous support medium, said sample chambers adapted to receive sample liquids, 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 secures to said base; a mounting plate assembly slidably guided by said posts and disposed laterally with respect to said sample plate and including, s\ r.a a mounting plate having4sldaby 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 57 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 movably 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 i. 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 sgnals 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 bir to aspirate a first predetermined amount nf 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 form 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, 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 S* application position where said microporous support t* 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 4 barrels, and an application position signal where ,o 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 micosyringe barLels 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 I \J l l O 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 san,,le 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 generatig a plunger bar lower position signal when 4":t said pluner actuator plate and said plunger bar are in said lo ,er position, and a plunger bar sample upper positon signal when said plunger actuator plate and sai, plunger bar are in said sample upper S position, and electrvcal 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, us- said mounting plate signalling means, and said plunger actuator plate signalling means, 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 t toperably generating and applying said up plunger 0 a actuator plate translation signal to said plunger actuator plate translating means thereby translating *said plungers from said lower position to said sample 0 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 c@ 61 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 sign'. when said application carriage position signal is received, said programmed digital computer means I *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 *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 microporuds support medium when secured to said lateral application space of said plate, wherein said sample plate further includes a wash well and a waste well longitudinally spaced from each other and from said sample chambers, t nd said plunger actuator plate traislating moans 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 4: plate assembly translation signal to said mounting plate assembly translating means thereby translating said mounting plate assembl, 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 I S t' 'WA 63 actuator plate translating means thereby translating said plungers f.'om 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 pr ,grammed 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, 0 40 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 s 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 1 I 64 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, and 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, t I said carriage translation means further i 4, includes a blotting position, and said carriage signalling means further includes means for generating a blotting carriage 0* j 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 ll 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 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 14 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, 4 I said motor having a rotatable output shaft, a pinion gear fixed to said output shaft, f a rack gear secured to said carriage, said rack gear being in engagement with said pii in gear whereby the output shaft turning of sdid motr translates said carriage. rC A' I 4N I 66

24. The apparatus of claim 17 wherein said signalling means for generating carriage position signals indicative uf 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 I \gitudinal blotting space, or said sample chamber, 4aid lateral application space is beneath said mi .owyringe barrels. 'o

25. The apparatus o' :laim 24 wherein said signalling means for generating carriage position signals indicative of carriage position inciAdes said carriage having a second longit'dinal surface with at least one notch disposed thereon near the forward and of said carriage, o a second trip switch secured to said trck on which said carriage moves, said second switch having a second spring forced roller ih'vans 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. ~r r, 67

26. The apparatus of claim 25 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.

27. The apparatus of claim 14 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 rot-table output shaft, pinion gear means fixed to said output shaft, rack gear secured to said base, said rack gear,4beg 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.

28. The apparatus of claim 14 wherein said signalling means for generating mounting plate p position signals indicative of mounting plate position includes ~upper stop means secured to a member secured to said base, So upper t cip 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 1 ran~ i 68 lower stop means for operatively tripping said lower trip switch means when said mounting plate reaches its lower limit of travel.

29. The apparatus of claim 17 wherein said means for translating said plunger bar up and down with respect to said mounting plate comprises a plngerr plate motor fixed to said mounting plate, said motor having a rctatable 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 t. said mounting plate.

The apparatus of claim 29 wherein said plunger actuator plate motor is fixed to the rear side of said mounting plate, said riounting plate includes two vertically plunger actuator plate guides with vertical grooves 44* provided therein, said guides being fixed to the n>o forward side of said mounting plate, said plunger actuator plate is slidingly disposed within said vertical grooves on said I 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 ~j-j 'i 1N~~ said mounting plate.

31. The apparatus of claim 29 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 ?lunger actuator plate, wash cam means vertically adjustably 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 tt 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 s 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 c*a T reaches its lower limit of travel. 0

32. The apparatus of claim 14 wherein said sample plate means further includes a row of individual liquid dilution wells longitudinally spaced from said sanple chambers, said diluted wells adapted to receive dilution liquid, said programmed computer means generating a Lit U A N St.. 0vr~ i i i: I 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 o- Sa\C\ ^Jsv c\XSr sa\AA SO-VNrFe_ combinationkof said dilution wells, and aspirating an amount of diluted liquid samples from said dilution wells into said respective pipette barrels.

33. Automatic pipetting apparatus comprising a base, sample plate means stationarily disposed on said base, said sample plate means having longitudinal and lateral limensions, said plate means including a lateral row of liquid sample chambers and 4 t 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 laterally above said sample plate means, longitudinal translation means for effecting longitudinal movement of said pipette frame above said stationarily disposed sample plate means, mounting plate carried by said pipette frame, vertical translation means for effecting relative vertical movement of said mounting plate and e? 'V o 71 said sample plate means, a plurality of microsyringe barrels having their heads secured in a -w to said mounting plate, said barrels spac' 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-plungsrs, each of said plungers disposed in one of said barrels, plunger translation means for moving said plpngers 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 mnunting 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 ior relative to said barrels, and programmed computer means responsive to said °o longitudinal signals, to said mounting plate signals and to said plunger signals for generating a sequence of control signals to said longitudinal translation 6 O. means, to said vertical translation means and to said 0 *s 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, 7 i I 72

34. An automatic pipetting apparatus substantially as hereinbefore described with reference to the accompanying drawings. DATED this 23rd day of October, 1989. HELENA LABORATORIES CORPORATION By its Patent Attorneys DAVIES COLLISON 14 4 QII I1 I I-