DE1798431B2 - DEVICE FOR THE INDEPENDENT DETERMINATION OF VARIOUS PARAMETERS OF A BLOOD SAMPLE - Google Patents

Description

Blood is made up of microscopic bodies suspended in serum. The predominant ones Corpora are called red blood cells, while white blood cells make up the lesser proportion. An examination of the blood characteristics requires both the examination the 'corpuscles or corpuscles per se as well as the whole blood, and for this purpose there has been since For some time a group of characteristic values or parameters which are recognized to be characteristic Are information carriers and provide the best possible description of a given blood sample. There is six particularly important parameters, all of which relate to red blood cells, their content, size, etc. relate. These parameters are used in the diagnosis, investigation, and treatment of anemia great importance. A reliable parameter that is primarily used to diagnose infections and with General health studies used refers to white blood cells.

The six most important parameters are: the number of red blood cells RBC (= red blood eel! count), the hematocrit index HCT, the hemoglobin HGB, the mean hemoglobin content MCH (= mear corpuscular hemoglobin), and the mean hemoglobin concentration MCHC (= mean corpuscular hemoglo bin concentration) and the mean particle volume MCV (= mean corpuscular volume). The seven " The parameter is the number of white blood cells WBC (= white blood cell count).

The first three parameters RBC, HCT and HGB as well as the number of white blood cells WBC are bishe ; tls parameter sizes have been measured directly, the other three parameters of red blood cells were true

Disher derived from or from the first three were calculated.

For a better understanding, the parameters and sizes are briefly explained in detail below.

RBC = red blood cell count

As the quantity corresponding to this parameter, the total number contained in 1 cc of blood becomes red Blood cells accepted. It is expressed in millions and is typically 5.5 million. According to the classical counting method, a blood sample was diluted, and a tiny amounts of this dilution were allowed to run into a counting chamber or into a hemocytometer. This chamber was of a known volume and meshed, and the technician had to Laboriously count bodies through a microscope and extrapolate the result obtained by multiplication with the dilution.

HCT = hematocrit index

The parameter represented by this value corresponds to the percentage of red blood cells on total blood volume. Because the corpuscles are elastic, disc-shaped cells with a liquid are inside, they can be compressed into a compressed volume under the action of centrifugal force. Usually the operator places the blood sample in a cylindrical chamber these rotate, as a result of which the corpuscles are deposited at one end of the chamber, and then represent the respective Amount of corpuscles and serum solid, the serum being an essentially clear liquid and without further above the dense red mass of the sedimented blood cells can be seen. Of the HCT index represents the percentage determined from this measurement. In the case of the present application The device shown here, the HCT index is not measured as a variable, but derived or derived as a parameter. calculated.

HGB = hemoglobin content

In medicine, this parameter is called the number of grams of blood pigment contained in 100 cm of whole blood Are defined. The fluid inside the red blood cells is an iron-containing complex protein compound, which gives the blood its characteristic red color. According to the known method To determine the hemoglobin, a lysine is added to a blood suspension Dissolves blood cells and exposes the hemoglobin. The free hemoglobin is then combined with one in the Reagent contained in diluent to react and gives a color-producing compound, which enables the determination. The color of the resulting solution is determined by colorimetric methods using any known device.

MCV = mean particle volume

This parameter is a measure of the average size of the corpuscles and is based on the normal average body size related. It is expressed in cubic microns. One compares the normal body size with the MCV of a blood sample to determine whether the average Size is a few percent larger or smaller. The MCV is usually made up of measured quantities calculated, namely by dividing the hematocrit index by the number of red blood cells, d. H. HCT / RBC.

MCI! - mean hemoglobin content of a particle

This parameter is a measure of what is contained in any blood cell regardless of its size Amount of hemoglobin. This is a number expressed in micromicrograms, ίο which is usually 29 micrograms and can be derived or calculated by division from HGB through RBC. Division by a normal value gives the coloring index.

NfCHC = mean hemoglobin concentration of one

Particle

This parameter is a measure of the hemogiobin concentration in an average body. For a given MCH, the following applies: the smaller the body. the greater the concentration. The value is in Percent and is roughly 3b percent. The size can be derived or calculated by Division of HG B by HCT.

WBC = white blood cell count

This is the number of white blood cells contained in 1 ecm of whole blood. Since that The ratio of red blood cells to white blood cells is more than 100: 1, the white will Blood cell count or WBC measured in units or in units multiplied by a thousand, where the Normal value is 5000. In order to do a white blood cell count, the red blood cell count must be Blood cells in the blood sample are excluded, and this is achieved again, as with the Hemoglobin determination, by a HämoKsc. at Use of a Coulter device in which the suspension passes through a microscopic passage opening must flow, it is important that thorough hemolysis occurs so that the resulting Decomposition products of the cell walls of the red blood cells lie within the "noise or interference range" on which the detector circles for the white Blood cells do not respond.

The three measured quantities or parameters are interrelated as explained above, and these interrelationships are known as blood indices. The three parameters MCV, MCH and MCHC are usually made up RBC, HCT and HGB are calculated and give the hematologist information about whether the corpuscles are large or small and whether the blood pigment is normal or abnormal or pathological. These indices provide also other important information.

DT-PS 9 64 418 describes, eitve particle detection device according to the Coulter principle. This makes it possible to determine the number of in a blood sample contained red and white blood cells and also the volume of these particles determine.

(, o Subject of the earlier German patent 16 73 146 is a device for the automatic determination of various parameters of a blood sample, with a sample dosing and distribution device for successive, mutually separated samples, at least one dispensing pre-

(, s direction for dilution liquid and / or reagents, Mixing devices, analyzing devices and evaluation devices which are used by the analyzing devices output parameters proportional

Measured values in proportional storable electrical Convert quantities and several of these electrical quantities for calculation can only be recorded indirectly Process parameters.

With this device, too, the hematocrit / .ahl, number of red blood cells, hemoglobin concentration, the mean corpuscular volume of erythrocytes, the mean corpuscular hemoglobin content and mean corpuscular hemoglobin concentration are determined. A number of these measured values are obtained through direct measurements and another number through Indirect determinations found. Here, however, no particle detection devices are found as analyzing devices use according to the Coulter principle. It is therefore not possible to use the count valuesc of the particles also determine the total volume of the counted particles. In addition, this older proposal the blood sub-samples one behind the other, only separated by air cushions, through the lines moved and the measured values of individual blood samples determined at the same time in the penetration process.

The present invention has for its object to provide a device which the automatic Determination anyway! the direct as well as the indirect blood measurement values are allowed in a simple manner and which, in contrast to the older proposal about the counted particles, also that Total volume of these particles can be determined.

The invention solves this problem with a device for the automatic determination of various parameters a blood sample, with a sample dosing and distribution device for successive, separate ones Samples, at least one dispensing device for dilution liquid and / or reagents, mixing devices. Analyzing devices and evaluation devices that are used by the analyzing devices submitted. Convert measured values proportional to parameters into proportional, storable electrical values and process several of these electrical quantities to calculate parameters that can only be measured indirectly, which is characterized in that the analyzing device used is particle detection devices are present according to the Coulter principle.

Advantageous further developments of the invention emerge from the subclaims.

The invention is described in greater detail below using an exemplary embodiment shown in the drawings explained. In the drawings shows

F i g. 1 is a schematic representation of the fluid system a device for the automatic determination of various parameters of a blood sample with block-shaped illustrated components,

F i g. 2 a table on which the control and duration of the individual Functions of the device and the operation of important parts of the device are shown; the table is vertically separated into two parts and consists of Fi g. 2a and F i g. 2b on two sheets.

F i g. 3 is a schematic diagram of the most important Components of the electrical circuit arrangement of the device: the circuit diagram consists of two parts, namely Fig. 3a and F i g. 3b on two sheets.

For a better understanding, the general structure of the device is first shown on the basis of the individual Functions explained.

The blood sample is taken in any way and in its entirety on a specific one Marking, usually in the form of a card with blank spaces into which the The information you are looking for is printed by the device's printer. The device comes with a snorkel type Provided tube, which is immersed in the blood sample and a certain amount in the fluid system soaks in. A tiny measured part of this whole blood is diluted with a predetermined amount transferred to a mixing chamber, where the first dilution then takes place. From this chamber

ίο comes together part of the suspension obtained with a pumped-in hemolysin in another mixing chamber, with the suspension in the second There is some time left in the mixing chamber for the red blood cells to dissolve and release their hemoglobin can.

A second part is withdrawn from the first mixing chamber and placed in another mixing chamber further diluted for red blood cell counts.

Each of the samples obtained is further treated separately after it has been produced. The White Blood cell sample, which contains the hemoglobin, is passed into a bath with three measuring tubes and by a constant liquid pressure in all three tubes simultaneously for a certain period of time sucked in. The measuring tubes are each provided with electrodes, and there is one in the bath common electrode so that when the white blood cells enter the tactile orifices containing measuring tubes three sets of signals are obtained. An electronic circuit provides one Detector output showing the number of white blood cells WBC directly. The White Blood cell samples with the hemoglobin they contain make it possible to determine the hemoglobin content HGB from the sample contained in the bath with the measuring tubes. It is accordingly Bath formed with a corresponding extension, which has parallel visible surfaces through which a beam of light is sent through the suspension and onto a photosensitive device and Provides information about the hemoglobin parameters of the original blood sample. One with the Output of the light-sensitive device connected electronic circuit results in the HG B representing Size.

Meanwhile the suspension of red blood cells is in a similar bath with measuring tubes. Electrodes and corresponding electronic circuits for detecting the occurrence of the suspension in the Measuring tube generated signals have been passed. A similar vacuum system or a pump takes care of it constant pressure, and as with the arrangement for the white blood cells, the palpation takes place over

a predetermined amount of time allowing a given volume of liquid to flow into all three Measuring tube corresponds.

The device also includes a device for filling and emptying the various containers as well for draining off already used and excess liquids. Once put into operation, the individual operations continuously, and the respective patterns do not contaminate each other.

The HGB mood as well as WBC and RBC provide three of the parameters through direct measurement. To the

MCV tuning is used to be on the safe side with two outputs from those obtained from the measuring tubes Red blood cell counts. That

System delivers analog electrical quantities, which these represent four parameters and store them in memory circuits.

Electronic means are provided for calculating the other three parameters. RBC and MCV are multiplied in a servo-driven computing device and result in HCT. HCT and HGB become in a similar device to give MCHC. MCV and MCHC are in the latter Device multiplies and yields MCH. The two derived parameters are MVHC, MCH and HCT also stored in order to be able to be printed on the card of the blood sample on demand.

The device is completely programmed via a series of control cams, which are arranged in a suitable rotary arrangement to one another on shafts rotating at constant speed. The control cams are simple switch actuation devices with cam surfaces which come into or out of engagement with the switches that are to be opened or closed in response to the movement of the control cams. These can be electrical switches or hydraulic or pneumatic valves. Since there is nothing unknown as such in this arrangement. no construction is shown in the drawings which shows the control cams, the cam drive or the switches or valves operated by them. F i g. Instead, FIG. 2 illustrates in the form of bars for which period of time which switches are opened and which are closed by the control cams. In FIG. 2, for example, sixteen control cams are indicated, each control cam being numbered accordingly. One set of control cams runs from C 1 to CS and the second set from C11 to C18. The control cam set is driven by a separate motor, which is specified in the table with timer 1 or 2. The first timer operates the control cams C1 with CS for a period of 15 seconds, as shown, and at the end of this period operates timer No. 2, but stops working itself. However, it can be restarted and as soon as it is restarted via a starter circuit, it goes through the same cycle again. Timer # 2 stops operating after 15 seconds and waits for the next signal from timer # 1. Although it takes 30 seconds to complete a full blood sample this way, the runs can overlap by fifteen seconds.

In Fig. 1 is the flow regulator for the exact measurement of the whole blood at the top left in the shown in a schematic representation and denoted by 10. It consists of three elements 12, 14 and 16, where the middle element 14 is arranged as an intermediate layer between the other two, however, as in FIG to describe individual, is pivotable in such a way that certain flow channels are aligned with one another will.

The middle element 14 is a construction manufactured with the highest precision, each with a single channel on opposite sides of a middle pivot point about which it can be pivoted. Each of these channels can take up a certain amount of a liquid and, by moving between two given positions, divide or close this liquid volume and pass it on or transfer it. This function is illustrated by arrows indicating the alignment of the central channels with the other guides in the upper and lower elements of the flow regulator 10. In this way, the upper element 12 and the lower element 16 are fixed to one another, and each element has four through openings. These passage openings in the upper element 12 are designated P1, P2, P3 and P4 and in the lower element with P5, P6, P1 and PS . When the middle part assumes its first position, its left channel P9 is aligned with the passage openings P1 and P5, while at the same time its right channel P10 is in communication with the passage openings P3 and P9. If the middle part 14 is rotated into its second position about its pivot point symbolically represented by the dashed line 18, the channels P9 and P10 move in the direction indicated by arrows into the dashed positions, ie according to FIG. 1 to the right, further flow being interrupted between the passage openings P1 and P5 and P3 and P7, while the channel P9 is brought into alignment with the passage openings P2 and P6 and the channel P10 with the openings P4 and PS.

This process can also be reversed and causes a certain volume of liquid from 2s of which one flow path can be removed and inserted into the other flow path, while the first flow path is blocked. For the sake of simplicity, the flow regulator is designed as a composite structure designated.

The following liquid lines are with the composite structure tied together:

1. Liquid line 20 leads from the passage opening P1 to the lower end of the blood sample control valve 22nd

2. Fluid line 24 connects flow port P2 to the top of the diluent control valve 26th

3. Liquid line 28 connects the passage opening P3 with the lower end of the dilution

central control valve 26.

4. Fluid line 30 connects flow port P4 to the top of the blood sample control valve 22nd

5. The fluid line 32 connects the channel P5 to the blood sample snorkel 34. Note that the snorkel in the diving position in a vessel 36 with a whole blood sample 38 dargestelli is. The vessel 36 is any container on which a certain identification is attached can be (not shown). Ready as above; mentioned, such a marking occurs arr best in the form of a card or the like, which can be inserted into a printer and which is through the Gera determined parameters for the respective blood sample is printed.

6. Liquid line 40 connects the Durchlauföff voltage P6 with the smaller chamber 42 de Mixing container 44 for the white blood cells.

7. Liquid line 46 connects the Durchlauföff voltage Pl with the smaller chamber 48 of the mixing container 50 for the red blood cells.

8. Liquid line 52 connects the passage opening 18 with the larger chamber 56 de Mixing container 44 for white blood cells. This line is also used as a removal device draws.

A blood sample pump 56 is connected to the blood sample control valve 22 via lines 5 and 60

509 587/41

ίο

and a diluent pump 62 via lines 64 and 66 to the control valve 26 for the diluent. Both control valves are three-way valves 22 and 26, the possible inner passageways being shown schematically in oblique dashed lines. The two flow paths in valve 22 are each designated with PIl and P11 'and in valve 26 with P12 and P12'. When valve 22, the average flow paths lead to the designated W discharge line. At valve 26, the central flow paths are connected to a line 70 of a source of diluent 52.

The pumps 56 and 62 are actually distribution valves with positive displacement pistons, which between move back and forth along the valve ends, displacing a volume of liquid. ) every pump picks up at one end the same volume of liquid that it dispenses at the other end.

Assuming the main flow regulator 10 is in the position shown in Fig. 1, wherein the center plate 14 is arranged so that the marked parts of the channels P9 and PIO are aligned on the left with the illustrated passage openings, so by actuating the blood sample pump 56 by moving their Piston from bottom to top - with the two passageways P11 'of the valve 22 open - a whole blood sample 38 is sucked into the line 32 and 20 via the channels Pi, P9 and P5. At the same time, any liquid located in the upper end of the sample pump 56 is discharged via the line 58 into the drainage line ^. During this time, the passageways PIl are blocked. The whole blood sample fills the channel P9, and as soon as the part 14 is pivoted to the other position, as shown by the dashed position of the channel P9, the blood remaining therein is brought into alignment with the passage openings P2 and P6.

The process described can be followed on the table in FIG. 2. The control curve CI here just closes a switch for a total of 15 seconds, as the bar 80 shows. This is a feed and self-holding function, so that the motor or the like. Drive device for the control cams continues to rotate during this entire period of time. The bar 82 represents the function of the control curve C1 , which is associated with the sequence of several operations. This actuation state affects the fluid system as shown by bars 82-1 to 82-6. The bar 82-1 represents the position of the central portion 14 of the composite structure 10, and position no. 1 corresponds to that in FIG. 1 marked position. Position no. 2 corresponds to that in FIG. 1 dashed position. The composite structure 10 is, as already mentioned, in its position No. 1 from the beginning to the end of the beam 82, that is approximately 1.5 seconds, the initial start-up time being considered to be zero. In fact, it takes a fraction of a second before everything starts, as the table shows.

The blood sample control valve 22 is immediately when the device according to the invention is put into operation in the position described above and is thus regulated in such a way that whole blood is poured into the Device sucks in. This is illustrated by bar 82-2. The sample pump 56 sucks, as described, blood in during the first 1/2 second, as illustrated by the solid bar 82-3, and promotes it up to the hatched bar 82-4, which shows that the lower end of the pump with Total volume is filled. As can be seen, the Solid bar 82-5 ', the the diluted sample in the upper end of the sample pump 56 from the first Represents passage to the hatched bar 82-5. which means that at the same time an emptying in the Drainage takes place.

The remainder of the bar table gives the state of the fluid system during the same period of time again. During the period represented by bar 82, the diluent control valve is located in the position shown at 83, which continues the position given before the start of the cycle. Of the Bar 84 shows that the diluent pump 62 is filled with diluent at the top Making the pattern for the white blood cell count, which it did. as the diluent for the red blood cell pattern.

For the dilutions to be made for red and white blood cell determinations, white must be Enter the order as the concentration is higher in view of the lower number of white blood cells have to be. The red blood cell pattern is made by diluting a beforehand for the white blood cells obtained dilution.

During the entire operating time of the first timer and the control cams Cl to C8 are certain parts of the fluid system become filled with fluid. Note the open bars 85.86 (two) and 88, which indicate the presence of previous patterns, for the Measuring tube immersion vessel 90 for the white blood cell sample which, for the sake of simplicity, is white bath is called; for the vacuum chamber 92. and for the measuring tube immersion vessel 94 for the red pattern, which is called the red bath for the sake of simplicity. These patterns come into play in the second half of the cycle Game.

The bar% starts at time 1.5 and ends around time 11, which means that it has a Represents a switch that is closed by the control curve C3 for a duration of 9.5 seconds. About that first half second of this state includes actuation of a device such as a relay. Cylinder Od. Like. A, which the middle plate 14 of the composite structure 10 in its second position sway. This is shown by the hatched vertical bar 96-1. For the rest of the bar% corresponding period of time, the composite structure 10 is constantly in position no. 2, as in 96-2 can be seen. The total amount of blood that lines 32 and 20 and the aligned Filled through channels through the composite structure 10 in position 1, is interrupted and blocked, and the small blood cone or plug caught in the channel P9 becomes with the passage openings P2 and P6 brought into alignment. As the top of the diluent pump 62 with diluent from the diluent source 72 was filled and the valve 26 on delivery of Diluent is set for the production of the white pattern, the line 22, which for Passage opening P2 does not carry any diluent after operation during this first half second of the pump 62 with the internal flow paths P12 'connected as shown and the flow

paths P12 are blocked. As represented by 96-3, the small plug of whole blood is transported by moving the central plate 14. This movement is represented by the vertical, downward-facing lines on bar 96-4, which is the white Specifying pattern.

Looking at the state of the various other bars, it can be seen at 97-1 that the same after completion of the movement of the center plate 14, the valve 26 switches to the position in which it releases diluent for the production of the white pattern, and in this position is the Line 24 not connected to the upper end of the diluent pump, its lower end on the diluent feed is connected. the This valve is regulated via the relay operated by the control cam C4 or the key cylinder. The bar 97 between the time 2 seconds and 5.5 seconds shows the actuation of a Mechanism which causes the valve 26 to be switched to the new position in which it is, as indicated by the Bar 97-1 shown until time remains 11.5 seconds. At the beginning of this period continued the delivery of the diluent as indicated by 97-2, with the delivery at time 5.5 Seconds is complete. Because at the same time with the promotion of the thinner for the white Samples from pump 62 to line 24 at the other end of the pump. Diluent from source 72 which ultimately becomes the diluent for the red pattern are bars 97-3 and 97-4 is shown at 1 to show that a Funding takes place. Once this pumping is complete, the end of the pump remains with the diluent for the red pattern until it is emptied on signal.

The plug of whole blood and diluent from line 24, represented by bar 96-3 pass through conduit 40 into the smaller chamber 42 of the mixing container 44 for white blood cells, and although just below the chamber ceiling and tangential to the chamber wall. The liquid stream flows invertebrate into the chamber. Before the blood plug in line 40 and after this in line 24 is located, as can be seen. Diluents. In this way, the circular movement causes the Tangentially entering liquid creates an invertebrate mixing movement in a horizontal plane. The entire in The amount of diluent flowing through the mixing container 44 corresponds to that displaced by the pump 62 Lot. These are 10 ecm, which push the blood plug into the chamber 42 and through flush the composite structure 10 connected through channels. The plug of whole blood is about 50 lambda, its exact volume being sufficient to provide the appropriate dilution at this point to give, which mixed with a hemolysin then results in a 250: 1 dilution.

The chambers of the mixing container 44 are completely empty when the paint begins to run in, whereby at 98 air enters the chamber 54 and flows through the chamber and the connecting channel 100 into the chamber 42. The control cam CA actuates the air valves to enable this process, the transfer of liquid into the chamber 54 being prevented until the point in time 5.5 seconds. This regulated air flow, which forms relatively large and slowly emerging bubbles, results in an invertebrate mixing movement in the vertical plane. The bluiprop is already well mixed with the diluent at this point since it is exposed to mixed components of relative movement in both the horizontal and vertical planes while the container 42 is filling. Once all of the solution has been poured in, there is a short delay, as shown by the end of the bar 97, and the control curve C5 actuates air valves, as shown by the bar 102, for a period of approximately 2.5 seconds in the reverse direction through the chambers 42 and 54. As a result, the pattern is conveyed through the line 100 into the chamber 54, and again tangentially against the chamber wall in order to largely exclude microscopic bubbles and eddies. The time interval between the reversal of the air flow also allows mixing by the moment of inertia. As with most fluid transfers in this device, the transfer rate is slowed from start to finish so that when the transfer is complete, the flow is practically zero. By adjusting the valves and actuators accordingly, this is carried out to a point where only three or four bubbles in the very last traces of the transferred liquid result in a complete emptying of the chamber, e.g. E. the chamber 42, ensure.

In the bar table according to FIG. 2 is the transfer from chamber 42 to chamber 54 represented by bar 97-5 going down to bar 102-1 is offset. After the transfer has taken place, the sample remains in the mixing container 44 for a certain time by the fully drawn bar 102-1 between the times 8.5 seconds and 9.5 seconds indicated, whereby the vesicles also subside.

At the point in time 9.5 seconds, the control curve Cb is put into operation and results in the bar 104, which extends over a period of 5.5 seconds up to the 15-second time mark. At the beginning of this period of time, i.e. at the point in time 9.5 seconds, the sample control valve 22 is moved into the position in which the inner passageways PI1 are aligned with the lines 58 and 60, whereby the line 30 with the throughflow openings P 4, PlO (at position no 2 of the composite structure 10), P8 and the line 52 is connected. which is immersed in the pattern located in the chamber 54. This setting of the valve 22 is represented by the bar 104-2. The sample pump 56 is now switched on and draws about 1 ecm of the liquid by movement; Plunger, flushing the opposite end, in which whole blood was located, into the drain line. The setting remains, and the opposite ending. ie the upper end is now filled with a thinned pattern which remains in it for the remainder of the first half of the cycle and also during the second half of the cycle until the new sample is introduced. In the illustration, the bar 82-4 is connected to the drawn-out bar 82-i, which illustrates the presence of whole blood in the lower end of the positive displacement pumps 56 until reprecipitation begins at the hatched part 82-4 of the bar between the points in time 8.5 seconds and 10 seconds, whereby the setting remains the same as at the beginning of the hatched part 82-4. The transfer of settings 82-3 through 82-5 includes a reversal of the settings at the left end of bar 82-7, s <

that the state in the ends of the sample pump is again represented by the bar 82-5 '.

A small plug of a diluted sample (one part of whole blood in approximately 250 parts of diluent) will collect in the P10 channel in setting 2. This transfer of a small pattern from the mixing container 44 to the composite structure 10 is represented by the vertical lines extending from the Extend bar 102-1 down to bar 104-1. At this point in time, the control cam CI again closes an actuating device in order to pivot the central plate 14 of the composite structure 10 back into its position 1. This shift is shown at 96-5 / between the time stamps 11 seconds and about 11.5 seconds. The condition at setting # 1 is again shown at 82-1 , with the bar 106 representing the operation of the control curve CI . Once the composite structure 10 is in position # 1, the diluent control valve 26 is actuated and placed in the position in which it dispenses the diluent for the red pattern. The control curve C1 now actuates the actuation medium for driving the pump 62, as illustrated by the bar 107.

During this movement, the entire amount of diluent (10 ecm) in the lower end of the pump is used to produce the red blood cell dilution, and this is from the transfer of the diluent at 1071 through line 28, passages P3, PIO and P7 and line 46 into the smaller chamber 48 of the mixing container 50 for the red blood cells can be seen. The same mixing process takes place here as in the case of mixing the white blood cells. The diluent flows from the control valve 26 through the described fluid channels and lines and pushes the plug of the already diluted sample with a volume of slightly more than 50 lambda out of the passage P10 and flushes it out. The path of this plug is indicated at 106-1 and extends down from bar 104-1 to bar 1C7-1. The setting of the pump 62 now changes from the bar 107-2, which shows the transfer of the diluent from the pump end with the red blood cells, via the bar 107-3, which represents a delivery, to the hatched bar 107-4, which shows that the end of the pump for the white blood cells is now filled with diluent. This condition persists for the duration of the entire bar 84 until the next sample is introduced. The setting of the dilution valve is again shown by the bar 83.

During this same time, namely from the point in time 11.5 seconds at which the return of the panel 14 of the composite structure 10 to its position no. 1 has been completed, the control curve CS is effective for the entire time corresponding to the beam UO. For the time shown there are two functions: First, the valve 112 in the drain line 114 is opened to allow the mixed white blood cell sample to enter the container 116, in which the red blood cells are dissolved. Up to this point in time, valve 112 was closed and valve 118 in line 120 was open. Second, a measured amount of a hemolytic agent is pumped from supply 122 through pump 124 and conduit 123 into container 116 of the pattern from container 44. The bar 110-1 represents the time during which the hemolysing agent is pumped, the bar being connected by vertical lines upwards to the bar 110-2 to show that the agent is entering the container 116. This is also running at the same time into the container 116 a pattern represented by the solid bar 110-3. as the vertical lines pointing downwards reveal me.

When transferring these fluids is always

ίο made sure that no bubbles appear. the

Inlet nozzles in the containers are aligned so that they direct their contents tangentially against the walls and the flow of liquid narrows or slows down will. This can be done by simple control pumps.

Reach cylinder and air pressure. When the

Fluid into the hemolysis reservoir 116 from the reservoir

44 air pressure can be used to ensure complete evacuation.

The last function performed by the first timer is indicated at 107-1, where the red blood cell pattern let into the first chamber 48 of the MiscliDehälters 50 wiro. as the bar 104-4 represents. That

The pattern remains in this chamber up to η time 18 seconds in the second half of the cycle Point in time, the degree of dilution is 5) 000 and is a function of the volumetric Ka timers and Passage channels of the device according to the invention.

At the point in time 15 seconds, the second timer is switched on by the first timer.

its operating time is then controlled by the control curve CU, as represented by the bar 125. The bars 126 and 132 also represent the function of various control curves Cl 2 to C18. The control curve C18, which operates the haemoj: lobin examination device, has two working periods, which are represented by two bars 132, which gives two readings which are identical to vt.

At a point in time of 15 seconds, no functions take place or have to take place in the device, those with the aspiration of the whole blood sample, the actuation of the composite structure and the movement of the sample and dilution pumps. Both patterns are even already mixed and ready for scanning. Hence, all are in the

_{The conditions prevailing <5} affected devices are the same as at the beginning of the first half of the cycle at time zero, as indicated by bars 82-1, 104-2. 82-5 '83 and 84 shown. As a result, a new sample can be introduced and the first half of the cycle initiated independently of the ongoing second half of the cycle. Appropriate shutdown circuits are used to : make restarting the first half of the cycle impossible until the control curve CIl has actuated a suitable circuit. Once the second half of the cycle has been initiated, it only stops when it is complete.

The only new parts of the device affected by the second half of the cycle depend on the running-in de Samples in their respective gauges and baths, taking measurements, rinsing, draining, etc. together.

The first function effected is controlled by the control curve C12, and the functions sin <are represented by the bars 126-1, 126-2 and 126-3. The first function is related to the measuring tube container 9 for the white blood cell pattern or the white bath, as it is called for the sake of simplicity. The bar 85 represents a previous pattern which was left in the white bath 90. To the

/ IU

At a time of 15 seconds, the valve 134, which is located in the line 135 coming from the flushing supply, is closed, the valve 136 in the line 138 is opened, and the previous lauster is drained from the white bath through a drain connection 140. which extends obliquely from the rinse supply to prevent any step back into the bath. This line 138 leads to a drainage container 141 which is controlled via a corresponding vacuum line 142. Recall that about 10 ecm of the pattern flowed into container 54; accordingly, the previous pattern in the bath 90 corresponds approximately to this amount, taking into account the addition of hemolyzing agent, the withdrawal for the counting, the liquid in the lines, etc. The measuring tubes 144 with the sensing openings 146 are arranged with a movable plate in such a way that they can be handled together. Its dimensions and those of the bath have been selected in such a way that 10 ecm of liquid carry the sample up to over the openings 146 through the volumetric space at the bottom of the bath.

Once the previous pattern into the drain line has been drained. as through the vertical ones, going down from the bar 126-1 to the thick ones Line 148, representing the drain line, the bath 90 is empty at the time 17.5 seconds. While at the same time, the previous pattern in the red bath 94, which corresponds to the bar 88, is in the Drain line drained as shown by bar 126-3, denoted by downward facing lines the second drain line 148, in the table below, is connected. This is the same type of devices in the Game, namely the valves 149 and 150, the lines 152 and 156, and the obliquely downwardly extending Discharge line 154. The red band 94 is constructed similarly to the bath 90 and has measuring tubes 158 with Touch openings 160.

The bar 126 also represents the conduction of the red blood cell pattern from the first chamber 48 in FIG the second chamber 162 of the mixing container 50 represents. The continuation of the beam 104-4 in the second half of the cycle shows this transition through the vertical lines to the bar 126-2, at which point the red pattern is already in the larger chamber 162. It stays in this chamber until it goes into the red Bad can be transferred, which takes place a little later in terms of time.

As soon as both containers have been emptied into the drain line, they are flushed out. this happens by closing valves 136 and 150, opening valves 134 and 149 and entering about 5 ecm Diluent in the bottom of each bath. This rinse doesn't quite go over that openings and takes place through the action of the control curve C13 during the through the bar 127 illustrated time. The left ends 127-1 and 127-2 of the bars on the baths give the running in of the Rinse thinner in the baths again. The control curve C14 controls the second emptying from the baths during time 128 as represented by right ends 128-1 and 128-2. As can be seen the Vacuum chamber 92 connected via line 170 and valve 172 to the drain container 141, and this Chamber 92 is drained simultaneously with the final emptying of the two baths. The beam 86 ends at time 21 seconds with vertical lines extending down to drain line 148 during

the time corresponding to the length of the bar 128.

The next function is controlled by the control curve C! 5 during the bar appropriate time. During this time, the white pattern is released through valve 118 and line 120 into his bath 90, while the red pattern is filled into his bath 94 via valve 174 and line 176 will. The first function is represented by the end of the bar 129 - 1, which merges into the bar 88. These both states, d. H. the baths filled with patterns remain that way. until the next pattern for counting are done.

The white pattern at this point consists of microscopic decomposition products of the outer cell walls of destroyed red blood cells, the intact white blood cells and the hemoglobin from the destroyed red cells. Taking into account the size of the passage P9 and the resulting plug (50 lambda), the 10 ecm diluent coming from the dilution pump 62, the sample taken from 52 prior to the hemolysis, the addition of hemolytic agent at 124 and the circulation in the device , the dilution of the white sample entering the white bath is almost exactly 250: 1. The red sample in the corresponding bath has a dilution of approximately 50,000: 1.

The counting process begins after a short settling time. This is via the vacuum chamber 92 to the Line 180 put a constant vacuum, a vacuum regulator connected at 182 is on a certain Value set while reading pressure gauge 184 what liquid is drawn from the baths into the measuring tubes during a given period of time and thus provides the volumetric pattern to be counted. This line 180 leads via the valves 186 and 188 to the separation chambers 190 and 192. Each of these chambers has three separate sections 194 each with a drip nozzle, which is connected to one of the measuring tubes of the corresponding bath. The sections 194 thus have drip nozzles 1%, which are connected to the corresponding measuring tubes 144 and 158 leading lines 1% and 198 are connected. If you place on chambers 190 and 192 a vacuum, the pattern is drawn through openings 146 and 160 into the interior of the respective tubes sucked in, and when the pattern passes through the openings it can according to the known Coulter principle are counted. The electrical connections and connections are not in Fig. 1, but in other figures shown. It should be sufficient at this point that each measuring tube has a separate inner Has electrode, and the bath for these tubes has a common electrode or ground electrode. By If the outlet drips off the lines 158, there is no electrical interference between the tactile opening circuits. The constructions for the white and red blood cells are both as shown formed similarly. However, the bath 90 only requires a rectangular extension 200 on its Floor. The same vessel saline can also be used for bath 94 for the sake of uniformity. From one Light source 202 is through optics 204. which have a green filter for generating monochromatic Including light of appropriate wavelength, a light beam sent through the extension 200 unc acts on the photocell of a hemoglobin determination

509 587/411

mation device 206. This takes place at the time 19.5 seconds by the control curve C 18, as the time bar 132 shows , so that. indicated in Figure 2 as in 132-1 below, which is used to rinse the bath 90, a measurement can be made of the pure diluent, and once / time to 29.5 seconds, the measurement 132-2 through the white pattern he follows. These data are used to calculate the hemoglobin parameter and are stored in appropriate circuits for the printer.

The counting process takes place simultaneously for both samples, as can be seen from the fluid transfers from the baths. The vacuum is applied for a significantly longer period of time than the actual counting process takes, by the action of the control curve C17, as the bar 131 shows. At the time indicated by the right end of the bar 85, the white sample is sucked into the measuring tubes, as indicated by the bar 131-1 , and from there into the vacuum chamber 92. where it remains for a certain period of time, w ^; e shows the bar 86 on the right. As far as the red pattern is concerned, the same process takes place as indicated by the bar 88 to 131-3 with 131-4 , up to the bar 86 on the right. The control curve C 16 initiates the count by momentarily turning on the circuit at 130 , the circuit remaining in counting mode for the time corresponding to the bar 130-1.

At the time of 30 seconds, all of the recorded data has been stored in an electronic circuit arrangement. This applies to both the measured and the calculated parameters. An automatic printing mechanism is provided which is actuated by inserting a card on which the data are to be printed. This is illustrated by bar 208 with all data being printed within 5 seconds.

3A and 3B show the most important components of the electrical circuit arrangement of the system, the control, reset and similar circuit connections having been omitted for the sake of simplicity. Since there are three measuring tubes rh "" ( for each thinned sample), six probe opening circles are provided, each with an amplifier, a threshold value circuit, an integrator, etc. for each probe opening Counting circles for the white blood cells are shown in the upper part of the circuit diagram and labeled with the letter W ; the counting and analyzing circles for the red blood cells are shown below the white circuits and labeled with the letter R. At the very bottom of the diagram, a hemoglobin circuit is drawn .

The white blood cell counting circles and red blood cell counting circles correspond, and therefore only the white blood cell circles need be described, with equivalent components and circuits having the same reference numerals. The differences are explained in the course of the description. In the upper left half of the circuit diagram, blocks 300-1, 300-2 and 300-3 represent the white tactile opening circuits which usually include the electrodes, circuit connections, etc. described above. The signals generated in these circuits result from the passage of the particles to be scanned through the scanning opening in

35

40

45

55

60 each measuring tube. The derived signals are used for amplifier 302-1. 302-2 and 302-3 placed, via which the current source 304 feeds all tactile openings. In practice, the openings have a diameter of approximately 100 μm and g <- ^ ... n ".." .. "" ^, """", so that one source of supply is sufficient for all. Current regulation is provided, and each individual channel can be adjusted accordingly in order to provide corresponding count values and signals.

The outputs of the amplifiers 302 are applied to the switching network 306 , which normally provides straight-ahead connections but enables cross-connections to be made during the adjustment periods. The output of each amplifier 302 is thus fed to its respective threshold circuit 308-1, 308-2 or 308-3. The amplifier outputs are also via the lines 310 together with the outputs 324 of the threshold value circuits to the cathode ray oscilloscope Kretse 312 so that each touch opening circuit on the os ^ illoscope 314 is assigned a picture or oscillogram. which reflects its operating status. By means of appropriate electronic circuits, the three oscillograms can be made equally visible as shown at 316 , and likewise the oscillograms of the key opening circles for the red blood cells, as shown at 318 . The majority of threshold circuits 308 are provided to integrators 320-1, 320-2 and 320-3 , respectively , each of which typically includes a capacitor, electronic switch, and diodes, discussed below

The pulses generated by the tactile opening circuits 300 must be converted into analog values to calculate the parameters measured in flour. These pulses are voltages at the threshold outputs 326-1, 326-2 and 326-3, provided the key opening pulse was large enough to exceed the set threshold value; each pulse gives a capacitor in the respective assigned pump circuit 322 Charge from, the latter gioi in turn from its charge in the connected integrator circuit 320 . The integrator circuit stores the charges and supplies a stored voltage proportional to the number of charge pulses and can be read at the respective outputs 328-1 or 328-2 and 328-3. The circuit is arranged in such a way that these outputs are medium as required! a switch SW- \ can be read which can be switched from the normal output of the election circuit 330 to each of the integration circuit outputs or to a measuring station. This enables the variable integration capacitors to be calibrated.

If all inputs of the constituency are in sthi have the same value, this means that all key openings are free and generate signals. Should be" Clog the opening, the signals coming from this opening differ significantly from the one Signals from the other two. The data is clogged through an electronic elimination process The first opening was eliminated and only the data from the other two openings were used. If all distinguish three signals significantly, they may result in all data being discarded.

Normally, the selector circuit 330 provides an output voltage which is the average of all dr < Inputs corresponds to; but should a Tastöffnun start to generate a signal that differs from de

nderen two by an amount dependent from the ^circuit: different actuator, it is automatically ausgechlossen and the other two used to determine the average value. The voltage level at 332 is attenuated in a Koir.zider.zk ^ correction-Net / -, vtrk 334, and the overall output is attenuated by a factor which is the conversion of the signal at 336 to a voltage equivalent to WBC. The output damper 338 regulates the size factor for properly printed responses. Two states are provided for the "> attenuator 340. In one state, the output of the coincidence / correction network 336 is passed through directly to 338 without any change. In the second state, a fraction of 336 and 332 'are added to create another coincidence- »5 to produce a correction factor which corresponds to a change in the size factor of the integrator 320-1, 320-2 and 320-3 The sense and purpose of this function will be explained later.

The number of white blood cells in a given blood sample is very small compared to the number of red blood cells; the fluctuations and dynamics are also very large. For the counting circles of the white blood cells, in which the count fluctuations are very large, would be the difference between the values generated by the integrators is very difficult while maintaining the stability of the generally available components to be processed. In populations with many wise men Blood cells should be the tension that each corpuscle at 328-1,328-2,328-3 represents less than that Voltage per pulse in sparse populations. Are the integrators and voting circuits set so that they provide a given output for a lower range, and then it turns out that the number white blood cells far exceeds the predetermined amount, then the circuit can be automatic be switched to compensate for this. This is preferably done by determining the state of charge in integrators 320.

A normally non-conductive size selector switch 342 receives from the integrator outputs 344-1,344-2 and 344-3 a voltage. As soon as one of the integrators is oversaturated, i. h "if the number of emitted pulses far exceeds the setting value of the switch, then a conductive state occurs one, and five circuits become conductive. Three of these circuits are controlled by line 346 and lead back to the integrators, adding additional capacitors to the integrating capacitors connected in parallel in order to change the size factor of the integrators. Two are in the Circuits 340 and 338 are provided and are fed by line 348. One changes the coincidence correction and the other the output damping to bring the scaling change into abutment. Of the The output of attenuator 338 is 350 and contains a DC voltage proportional to WBC.

Since with the red blood cells the numerical fluctuations are not as great as with the white ones Blood cells, these fluctuations can be easily handled by an integrator scale will. Accordingly, there is also no equivalent circuit for the red blood cells in the circuits the switch 342 and its associated part are provided. In all other respects the counting circuit is for the red blood corpuscles with that for the white niiitlfisrnerchen identical in structure, and the corresponding Components are marked with the same reference numerals. To the circuit arrangement for the red blood cells are usually more stringent than those for white blood cells the size of the red blood cells is often determined. Accordingly, the amplitudes should be if possible, are retained up to the point in time at which the size information corresponds to the original Signal channels can be taken. In this case, since MCV is obtained, the size information becomes direct taken from amplifiers 302-1 and 302-2. Because a dial 330 is provided in the red part of the device. the use of two outputs on 352 and 353 ensures greater reliability wedge for MCV determination, if one of the channels is blocked. These signals are sent to the MCV attenuators 354-1 and 354-2 supplied and forwarded from there to the MCV supplying device 356-1 and 356-2.

The outputs of the MCV devices are applied to averaging means in selector circuit 330 via lines 358 so that the output at 360 provides an average signal proportional to MCV. Bypass lines lead to the poles of the Shahs SW-2, which is operated in conjunction with the constituency to turn off one of the lines 358 and the MCV limit switch if the result indicates that a line is providing incorrect information. This circuit allows the same voltage factors at 360 once the average of the two MCV signals or, in the case of a clogged orifice elimination, either is used. A correspondingly attenuated value at 362, which is proportional to the MCV and is converted into the corresponding value for the actual MCV, is output to the line 364. Two further outputs can also be derived from the MCV information, one being attenuated by a different amount at 366 to yield the voltage at 368 which is a function faMCV of MCV, and the other is not attenuated on line 370 and represents a function fbMCV.

RBC of the red blood cell count is provided by attenuator 338, which is the white attenuator 338 corresponds to the size factor circuit to which Line 372 released.

The hemoglobin measuring device 206 is shown below in Figure 2a. The photosensitive device 206 is shown in FIG. 1 illustrates and provides a current that is amplified in amplifier 374 and in the computer 376 is converted into a voltage at 378, which after being damped or damped accordingly at 380. has been weakened, at 382 results in a size analogous to the HGB of the blood sample. This can be converted into a digital size can be converted. The signal emitted at 378 is in the vapor; 384 in his Scale changed to provide a different voltage on line 386 for use in the calculation of the already mentioned indices. This value is a function of the HGB of the HGB.

As can be seen, the RBC voltage at 336 with no attenuation is used in another calculation and on line 338 to one of the computing devices. This value is a function of fRBC RBC.

The information measured and to be calculated in the device is printed on a card or on otherwise transferred to vision devices or recorders. This requires a conversion

The analog information is converted into digital data, but for the sake of simplicity the signals are initially treated as analog quantities. The lines 350,364,372 and 382 lead directly to the distribution device, namely the commutator 390. Each of the seven parameters has a pole, and the commutator scans all the poles in turn and transmits the information to the printer 392 in the appropriate order has converted digital data in the converter 394 . The information can optionally be sampled in a corresponding device 396 in order to obtain an indication of the analog quantities for other purposes in the line 398. The printer drive 400 advances the card each time. when a parameter has been recorded for a given run.

The computer 402 supplies HCT and the computer 404 supplies the other two indices.

The voltage labeled fRBC is proportional to RBC and is applied via line 338 to servo amplifier 406 , which drives motor 408 , which in turn rotates a wiper 410 on a potentiometer 412 which is set in this way. that there is a predetermined voltage to earth across it. This voltage is obtained from an appropriate reference source and adjusted to the required value by a divider 414. The position of the wiper 410 is thus related to RBC, and the feedback of the error voltage through line 416 keeps the error voltage at zero by rotating it accordingly. There is a voltage across potentiometer 420 which is proportional to MCV. because line 370 is connected at its higher potential end. As mentioned earlier, the voltage on this line is a function faMCV of MCV. Since the rotation of the grinder is 410 with the slider 422 in synchronism, the voltage at 422 is equal to the product of the voltage on the higher potential having the end of the potentiometer 420 and the rotation or RBC, and this product is directly proportional to HCT . Therefore, the output is the voltage stored at the tip of wiper 422 and appears on line 424.

The voltage on wiper 422 is amplified at 426 and applied via line 428 to the tip of potentiometer 430 so that the voltage is above

ίο this potentiometer is proportional to HCT. This takes place in the computer 404 and, as can be seen, two other potentiometers 432 and 434 are also provided. Potentiometer 432 is connected to line 368 so that the voltage faMCV on the line is proportional to the voltage across the potentiometer

MCV lets become. There is a fixed voltage across the potentiometer 434 , which voltage is determined by the divider 436 and the value of the reference voltage.

In the same way, the servo amplifier arrangement described above brought the veil 410 into its rotational position proportional to RCB, so that the servo amplifier 438, the motor 440, the feedback line 442 and the connection of the servo amplifier 438 to the line 386 cause the rotational position of the Grinder's 444 is proportional to HG B divided by HCT or to MCHC. This results from the fact that instead of a constant voltage across the potentiometer 430, such as. B. in the case of the potentiometer 412, the HCT voltage is applied, and this is variable.

The feedback 446 provides constant zero adjustment.

The last quantity received, namely MCH. lies above the wiper 452 of the potentiometer 432. Since the position of the wiper 452, which is synchronized with the other wipers, is proportional to MCHC. and since the voltage across potentiometer 432 is proportional to the MCV. MCV is multiplied by MCHC, and then on line 454 is MCH.

In addition 5 sheets of drawings

Claims (8)

Patent claims: 1. Device for the automatic determination of various parameters of a blood sample, with a sample dosing and distribution device for successive, separate samples, at least one distribution device for dilution liquid and / or reagents, mixing devices, analyzing devices and evaluation devices which are delivered by the analyzing devices. Convert measured values proportional to parameters into proportional storable electrical quantities and process several of these electrical quantities to calculate parameters that can only be recorded indirectly, characterized in that two separately working, different parameter determining particle detection devices (144, 146, 158, 160) according to the Coulter principle are used as analyzing devices available. 2. Apparatus according to claim 1, characterized in that each Coulter analyzing device (90, 94) via a drip connection (196) to a chamber (190, 192) and this via a controllable valve (186, 188) to a vacuum source (92, 184 ) is connected. 3. Apparatus according to claim 1, characterized in that each Coulter analyzer has a plurality of measuring openings of the same size and separate detector devices are connected to each measuring port, that a selector circuit is provided for each Coulter analyzer and is separately connected to each detector device of the analyzers and that a Oscilloscope (314) is present, which is connected to each detector device for simultaneous display for the oscillograms (316, 318) generated by all detector devices. 4. Apparatus according to claims 1 to 3, characterized in that a Teüchenkoincidence correction circuit (334) is available for the measured quantities analogous to the particle signals, that a stable multivibrator (338) is connected to the output of this correction circuit and that one on the change the dynamics of the stored and converted signals responsive circuit (342) for changing de. Coincidence and the scale of at least one of the analog quantities is present. 5. Apparatus according to claim 1, characterized in that a hemoglobin measuring device (206) for generating an analog electrical measured variable which is proportional to the hemoglobin in the once diluted partial sample is connected to the one Coulter analyzing device (90). 6. Device according to one of the preceding claims 1 to 5, characterized in that a computer (402, 404) is available as an evaluation device for recording and combining the determined number of red blood cells, the hemoglobin and the mean particle volume, which supplies analog electrical quantities which are proportional the average hemoglobin content and the average hemoglobin concentration of a body and the hematocrit index. fts 7. Apparatus according to claim 6, characterized in that the computer includes an arrangement (406 ...) for multiplying the analog values of the mean particle volume and the number of red blood cells to derive an analog value proportional to the hematocril index and that furthermore a second arrangement (432) to multiply the analog size proportional to the minimum particle volume by the analog size proportional to the mean hemoglobin concentration of a body and to derive an analog electrical output which is proportional to the mean hemoglobin content of a body. 8. Apparatus according to claim 7, characterized in that the computer (404) contains a servo amplifier, with a first potentiometer (430) with a grinder (444). which applies the analog electrical quantity proportional to the hematocrit index to the first potentiometer, the grinder causing the error voltage to be fed back so that a second and third potentiometer (434) each with a rotating grinder is present. A reference voltage is applied to the third potentiometer so that its wiper decreases the analog electrical quantity proportional to the mean hemoglobin concentration, the analog electrical quantity proportional to the mean particle volume being present over the second potentiometer and the wiper the analogue electrical quantity proportional to the mean hemoglobin content proportional electrical quantity decreases.