JPH01212363A - Method for supplying blood or the like - Google Patents

Abstract

PURPOSE:To obtain a method for preventing the lowering of analytical accuracy due to the scattering of a liquid due to coagulation, by using a pipet capable of discharging a definite amount of a liquid once or repeatedly as a liquid metering container. CONSTITUTION:Chemical analysis slides 1a-1f are respectively received in the heat insulating chambers 11a-11f provided to an incubator 10 held to definite temp. by a heater and the coloration of a reagent layer is measured as reflected optical density by an optical measuring probe. A slide feed means 50 having the slides 1a-1f mounted thereon is moved along the heat insulating chambers 11a-11f to be inserted in and discharged from the heat insulating chambers 11a-11f by a lever 53. When the means 50 is positioned at the left- hand edge of a stator 52, the center each of the spot deposition orifice of each of the slides 1a-1f is positioned directly under the nozzle of a pipet 60 to deposit whole blood in a spot form. At this time, the outer diameter of the leading end of the nozzle is set to 2mm or less and the liquid sucked in the pipet 60 is discharged at flow velocity of 60mul/mm or less per a sec and the liquid is repeatedly supplied in a definite amount of 20mul or less.

Description

[Detailed Description of the Invention] [Field of Application of the Invention] The present invention provides a chemical analysis element, for example, a chemical analysis slide, in which:
This invention relates to a method of dot-supplying a fixed amount of a sample liquid using a liquid injector, a dispenser, etc.

[Prior art and its drawbacks] When quantitatively analyzing a specific chemical component in a biological fluid such as blood or urine using a chemical analysis element, such as a chemical analysis slide, a predetermined amount of sample liquid must be measured with the required precision. Dispensers used for this purpose, which must be metered and supplied to the analytical slides, are widely used, for example, from Eppendorf (e.g. No. 4780) and Nichiriyou (e.g. Model 8100). ing.

Blood, plasma, etc. begin to coagulate after about 20 seconds when exposed to air. If a dispenser with a fine-tipped nozzle is used to supply such a liquid sample with coagulability, the liquid sample (hereinafter referred to as the sample) can be inhaled and discharged, or after being discharged for - times. If the sample is left for a short time before being discharged, the sample will coagulate at the tip of the nozzle, making it difficult to discharge the sample. For example, at a temperature of 24°C and a relative humidity of 40%.
Under these conditions, if the sample is blood, such coagulation will occur in about 40 seconds. Although it will not be difficult to drain the sample in about 20 seconds, in the case of zero-reaction cuvettes, the removed liquid will scatter and the accuracy of the amount of liquid supplied to chemical analysis slides, chemical analysis tapes, reaction cuvettes, etc. will decrease. The specimen adheres to the wall surface, and on the chemical analysis slide or chemical analysis tape, the specimen adheres to areas other than the predetermined areas and is not used for analysis, resulting in a decrease in analysis accuracy. In chemical analysis slides and chemical analysis tapes, the sample does not adhere uniformly to the predetermined area required for quantitative analysis.
Analysis accuracy decreases. Therefore, in order to supply such coagulable specimens by means of a filler or dispenser, the time interval between aspiration of the specimen and the start of evacuation or repeated evacuations should be relatively short, i.e. within about 15 seconds. There was a need.

However, when dispensing a single sample, the interval between discharges may unavoidably exceed 15 seconds.Dispenser dispensers are often used in combination with automatic analyzers, and in such cases, the dispensing of the sample is not necessarily at the same time interval. The automatic analyzer performs the chemical reactions necessary for the analysis, which may not necessarily be repeated, so the ink.

If the time required to pass through the incubator is fixed, which is often equipped with a beta (constant temperature room), it is possible to pass the analytical slide, analytical tape or analytical cuvette continuously through the incubator; In general, sample dispensing is performed continuously. However, the dispensing of the sample is not necessarily carried out continuously.
When using an automatic analyzer, such as the automatic analyzer described in No. 94368 (Sugaya), which is equipped with an insulating chamber that individually houses the analysis slides and can keep each slide at a constant temperature for an arbitrary period of time. In order to efficiently perform analyzes (as many as possible within a certain period of time), it is important to keep the insulated chamber as empty as possible, so when the first sample is dispensed, If there is space in the room, the second
Start the analysis by dispensing (supplying) the first sample onto the analysis slide, etc., wait until the analysis of these samples is completed and the incubation room is empty, then dispense the third and subsequent samples onto the analysis slide, etc. In such a case, between dispensing the first sample and dispensing the second sample, it may take some time, for example, 15 seconds or more, to clean or replace the nozzle of the dispenser and aspirate the second sample. time is required, so the time interval during which the incubation chamber is empty is also shorter than when samples are not exchanged (1
(0 seconds or less), and the time interval during which the insulating chamber is empty after the analysis is completed is uneven. Therefore, the third
During sample analysis, the intervals between sample dispensing (which must be done after waiting for the incubation chamber to become empty) become uneven, and the time (approximately 15 seconds) that may cause the sample to coagulate may be exceeded.

Furthermore, it is often necessary to wait for 15 seconds or more to prepare the analysis slide to be spotted. In such a case, coagulation of the sample reduces the accuracy of the amount of liquid supplied to the analysis slide or the like, or the scattering of the liquid reduces the accuracy of analysis.

- Even with injectors that drain only once, it is often necessary to wait 15 seconds or more until the analysis slide or incubator is ready, reducing analysis accuracy.

[Technical Problem to be Solved] The technical problem of the present invention is that in a method for supplying a fixed amount of a coagulable sample liquid using a liquid injector equipped with a fine-tipped nozzle, it is difficult to aspirate a sample. It is an object of the present invention to provide a method for supplying a sample liquid in which analysis accuracy does not deteriorate due to scattering of the liquid due to coagulation even if the time from start to discharge unavoidably exceeds 15 seconds.

The present invention also provides a method for repeatedly supplying a fixed amount of coagulable sample liquid using a dispenser equipped with a fine-tipped nozzle, in which the interval between discharging the sample liquid unavoidably exceeds 15 seconds. We also aim to provide a sample liquid supply method that does not reduce analysis accuracy due to liquid scattering due to coagulation.
Consider it a technical issue.

[Means for solving the technical problem] The above problem is such that the outer diameter of the tip of the liquid discharge nozzle is 2 mm or less, and the liquid sucked into the injector or dispenser is transferred from the liquid discharge nozzle at a flow rate of 60 μm/lll112 or less per second. The problem was solved by a liquid supply method, which is characterized by discharging the liquid at a rate of 20 μl and supplying a fixed amount of liquid of 20 μl or less once or repeatedly.

The present invention is particularly effective when supplying whole blood, whole blood diluted solution, plasma, or plasma diluted solution using a syringe or dispenser.

[Specific structure of the invention] The liquid supply method of the present invention usually includes a liquid measuring container,
The method of the invention, in which a pipette is used which can dispense a fixed volume of liquid once or repeatedly, is particularly useful for delivering liquid volumes from 2 μl to 20 μl.

Many pipettes have a cylinder and a piston.

Further, the nozzle may be fixed, or a removable and disposable nozzle tip may be used. The method of the invention includes, for example, U.S. Pat.
1, 3,732,734, 3,732,735, 3,757,586, 3,766.784, 3,7
66.785 and 4.023,716 can be used. In addition to the above-mentioned commercially available nozzle tips, for example, U.S. Pat.
, 237,095 and 4.347,875 can be used. The surface of the nozzle is US Patent 3
．． 500,689, commercially available nozzle tips, which may be coated with a hydrogenated hydrocarbon polymer, are often made of polypropylene due to processing accuracy.

The outer diameter of the nozzle tip of the liquid injector or dispenser such as a pipette used in the method of the present invention is about 0.31 to 2 mm. Preferably it is 0.4 mm to 1.61 mm.

When used in the method of the present invention, the inner diameter of the tip of a nozzle such as a bed is about 0.2 am to 1.4 m11+.

Preferably it is 0.3 mm to 0.8 n+m.

The length of the nozzle naturally depends on the desired amount of liquid to be accommodated, since in the case of a disposable tip, the entire amount of liquid is accommodated.

[Effect of the invention] When a fixed amount of a coagulable sample solution such as whole blood, plasma, or their diluted liquid is supplied once or repeatedly using a liquid injector or a dispenser, the inhalation of the sample Even if the time to drain or the time interval between drains unavoidably exceeds 15 seconds, the accuracy of analysis will not deteriorate due to scattering of the liquid due to coagulation.
Chemical analysis slides and chemical analysis tapes that do not cause the specimen to adhere to areas other than the designated area, and in reaction cuvettes, the specimen does not adhere to the walls and are not used for analysis, resulting in a decrease in analysis accuracy. Also, in the chemical analysis tape, there is no decrease in analytical accuracy due to the specimen not being uniformly attached to a predetermined area required for quantitative analysis.

[Example: An automatic analyzer was manufactured according to the description in JP-A-61-294368. Figure 1 shows its front view.

The incubator 10 is kept at a constant temperature by a heater (not shown), and is a 1 lb. incubator.

Equipped with 11c, lid, lie, and llf. Each of these greenhouses contains chemical analysis slides la.

1b, Lc, ld, le, and if are stored. An optical measurement probe 30 is provided below the incubator 10,
It is possible to move along the rows of the insulating chambers 11a to llf, and therefore along the rows of stored chemical analysis slides 1a to 1f. Measurement openings 12a to 12f are provided below each of the insulating chambers 11a to llf, and through these openings 12a to 12f are provided, through which the optical measurement probe 30 measures the color development (color change, fluorescence, etc.) of the reagent layer of the chemical analysis slide as reflected optical density, etc. can be measured.

The chemical analysis slides are sequentially transferred to the slide transport means 50.
It is placed on top of the . The slide conveyance means 50 is provided with a stator 52 or is moved by a near motor along the rows of the insulating chambers 11a to llf, and the chemical analysis slides 1a and the like are inserted into the insulating chambers 11a to llf by a lever 53. The chemical analysis slides that have undergone the necessary reactions and measurements in the incubation chamber are discharged from the incubation chambers 11a to llf by further inserting the lever 53 deeper.

When the slide conveying means 50 is located at the left end of the stator 52, the outer diameter is 0.65 mm and the inner diameter is 0 so that the center of the dot on the chemical analysis slide is located directly under the tip of the nozzle.
．． Install a dispensing pipette (of the type described in Japanese Patent Application No. 61-144258) with a total volume of 110 μl using a 5 mm nozzle tip.

4N below! Chemical analysis slide 1a for test items in J
Immediately after aspirating μl, the chemical analysis slide 1a is placed on the slide transport means 50 located at the left end of the stator 52, and the first whole blood sample 10μe is transferred at 8 μm (4 μm) per second.
After the spotting, the nozzle tip is filled with the sample liquid up to the tip.The slide conveying means 50 is moved over the stator 52 by a linear motor into the insulating chamber LLa. Move it all the way to the front and press lever 5.
3, the chemical analysis slide 1a was inserted into the incubation chamber 11a. Return the slide conveyance means 50 to the left end position of the stator 52, and similarly spot whole blood on the chemical analysis slide 1b.
It was inserted into the greenhouse 11b.

The spotting on slide 1b was done 10 seconds after the spotting on slide 1a. Continue in the same manner for chemical analysis slide 1
Whole blood was spotted on slides c and 1 d sequentially and inserted into the incubation chamber, but the spotting on slide 1c was 30 minutes after the spotting on slide 1b.
Seconds later. The spotting on the slide 1d was done 10 seconds after the spotting on the slide IC.

Replace the nozzle tip of the dispensing pipette with a new one, aspirate the second whole blood sample, and apply 10 μl each of the second blood sample to chemical analysis slides 1e (same type as la) and 1f (same type as lb) in sequence. 11 e and 11 respectively.
After the liquid was spotted on the zero analysis slide 1f inserted in the slide 1f, the tip of the liquid was retreated to 1 mm from the tip of the nozzle tip within 3 seconds.

Approximately 20 minutes after inserting slide 1d until inserting slide 1e
It took seconds.

After keeping the chemical analysis slides 1a to 1f warm for 6 minutes in each of the insulating rooms 11a to llf,
Measuring openings 1 provided below each of
The reflected optical densities of the reagent layers of the chemical analysis slides 1a to 1f were sequentially measured using an optical measurement probe 30 that was movably provided below the incubator 10 through 2a to 12f. After the optical measurements were completed, each slide was discharged outside the incubation room.

The remainder of the second whole blood sample is retained in the nozzle tip of the dispenser for approximately 7 minutes until the slides 1 a and 1 b in the incubation chambers 11 a and 11 b are discharged, and then transferred to the slide transfer means 50 at the spotting position. It was spotted onto 1 g of a chemical analysis slide placed on top of the glass. After inserting the slide 1g into the insulating chamber 11a, dotting was performed on the slide 1h in the same manner. Slide 1g
The time interval between dotting and 1 h was 20 seconds. Slide 1
h was inserted into the insulating chamber 11b immediately after spotting. Slides 1g and 1h are each 6mm in the same way as slide 1a etc.
After minutes, the reflected optical density was measured and the chamber was then discharged.

The whole blood sample and the first and second samples were also dispersed without scattering.
It was successfully spotted approximately circularly on the analysis slide. For comparison, the discharge speed was set to 30μp/s (150μ1ists2
), when the blood was spotted on the analysis slide IC, the blood was scattered and spotted on the analysis slide, causing an error.

[Brief explanation of the drawing]

FIG. 1 is an elevational view of an automatic analyzer used in an example of the present invention, and FIG. 2 is a plan view thereof.

Claims (1)

[Claims] 1) In a method for supplying a fixed amount of coagulable liquid using a liquid injector having a liquid discharge nozzle, the outer diameter of the tip of the nozzle is 2 mm or less, The aspirated liquid was discharged at a flow rate of 60 μl/mm^2 or less per second, and
characterized by supplying a certain amount of liquid of less than μl;
Liquid supply method. 2) In a method for repeatedly supplying a fixed amount of coagulable liquid using a dispenser having a liquid discharge nozzle, the outer diameter of the tip of the nozzle is 2 mm or less, and the liquid sucked into the dispenser is Discharge at a flow rate of 60 μl/mm^2 or less per second,
A liquid supply method characterized by repeatedly supplying a fixed amount of liquid of 20 μl or less. 3) The liquid supply method according to claim 1) or 2), wherein the sample liquid is whole blood, whole blood dilution, plasma, or plasma dilution. 4) The time interval of liquid discharge exceeds 15 seconds at least once during repeated feeding.
) liquid supply method.