U.S. patent application number 12/845130 was filed with the patent office on 2011-03-03 for specific component measuring method by spectral measurement.
This patent application is currently assigned to WAKO PURE CHEMICAL INDUSTRIES, LTD.. Invention is credited to Wataru AKAHANE, Takashi FUJITA, Hiroyuki YAMADA, Sachiko YAMAMOTO.
Application Number | 20110052448 12/845130 |
Document ID | / |
Family ID | 33566804 |
Filed Date | 2011-03-03 |
United States Patent
Application |
20110052448 |
Kind Code |
A1 |
FUJITA; Takashi ; et
al. |
March 3, 2011 |
SPECIFIC COMPONENT MEASURING METHOD BY SPECTRAL MEASUREMENT
Abstract
The present invention discloses a method for measuring an amount
of an objective component to be measured in a sample, which
comprises; preventing an electric charge in an atmosphere in a
photometry chamber from transferring to the surface of a solution
which generates light due to an energy variation of a substance
induced by the objective component in the sample, measuring value
of the light, and determining an amount of the objective component
in the sample on the basis of the measured value thus obtained, and
an instrument used for the method. According to the present
invention, in measurement of an objective component in a sample
using a spectrophotometer, problems such as between-day variation
of signal values or increase of background value, etc. can be
solved, and a trace component can be measured in high accuracy and
high sensitivity.
Inventors: |
FUJITA; Takashi; (Hyogo,
JP) ; YAMAMOTO; Sachiko; (Hyogo, JP) ; YAMADA;
Hiroyuki; (Hyogo, JP) ; AKAHANE; Wataru;
(Hyogo, JP) |
Assignee: |
WAKO PURE CHEMICAL INDUSTRIES,
LTD.
Osaka
JP
|
Family ID: |
33566804 |
Appl. No.: |
12/845130 |
Filed: |
July 28, 2010 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10561538 |
Dec 19, 2005 |
7795034 |
|
|
PCT/JP04/08711 |
Jun 21, 2004 |
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12845130 |
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Current U.S.
Class: |
422/82.08 ;
422/82.05 |
Current CPC
Class: |
G01N 2201/023 20130101;
G01N 21/76 20130101 |
Class at
Publication: |
422/82.08 ;
422/82.05 |
International
Class: |
G01N 21/64 20060101
G01N021/64; G01N 21/00 20060101 G01N021/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jul 3, 2003 |
JP |
2003-270797 |
Mar 31, 2004 |
JP |
2004-106577 |
Claims
1. An instrument for measuring value of light generated due to an
energy variation of a substance in a solution instrument, which
contains a mechanism for measuring value of light generated due to
an energy variation of a substance in a solution and a mechanism
for preventing an electric charge in an atmosphere in a photometry
chamber from transferring to the surface of the solution, wherein
the mechanism for preventing the electric charge in the atmosphere
in the photometry chamber from transferring to the surface of the
solution is a mechanism for making an atmosphere surrounding a
reaction vessel and/or an atmosphere surrounding the reaction
vessel in the photometry chamber electrically constant, and is one
or more mechanism selected from the group consisting of: (1) an
apparatus for flowing gas having a constant electric charge to the
atmosphere surrounding the reaction vessel and/or into the
photometry chamber, (2) a neutralization apparatus, or (3) a
photometry chamber on which a material having a static electricity
elimination effect is attached.
2. The instrument according to claim 1, wherein the light generated
due to the variation of the energy of the substance is
luminescence, fluorescence, or phosphorescence, which is generated
corresponding to a presence of an objective component to be
measured in a sample.
3. The instrument according to claim 1, wherein the mechanism for
making the atmosphere surrounding the reaction vessel and/or the
atmosphere surrounding the reaction vessel in the photometry
chamber electrically constant is two or more mechanisms selected
from the group consisting of: (1) an apparatus for flowing gas
having a constant electric charge to the atmosphere surrounding the
reaction vessel and/or into the photometry chamber, (2) a
neutralization apparatus, or (3) a photometry chamber on which a
material having a static electricity elimination effect is
attached.
Description
CROSS REFERENCE TO RELATED APPLICATIONS
[0001] This application is a divisional application of Ser. No.
10/561,538, filed Dec. 19, 2005, which is a National Stage
Application Under 35 U.S.C. .sctn.371 of PCT/JP2004/08711, filed
Jun. 21, 1994, which is based upon and claims the benefit of
priority from the prior Japanese Patent Application No.
2003-270797, filed Jul. 3, 2003 and Japanese Patent Application No.
2004-106577, filed Mar. 31, 2004, the entire contents of which are
incorporated herein by reference.
TECHNICAL FIELD
[0002] The present invention relates to a method for measuring a
trace component in a sample originated from a living body such as
serum, plasma, or the like using a measuring instrument. In
particular, the present invention relates to a method for
quantifying an objective component to be measured in high accuracy
by preventing influences such as measurement errors or increased
background value caused by localized electric charge in measuring
environment, especially in an atmosphere surrounding a reaction
vessel, while measuring luminescence, fluorescence,
phosphorescence, or the like using a spectrophotometer, and an
instrument used for the method.
BACKGROUND ART
[0003] Analysis and/or measurement of an objective component to be
measured by measuring luminescence, fluorescence, phosphorescence
or the like have been widely utilized especially for the
measurement of trace components such as environmental hormones or
in vivo substances, because of its high sensitivity. In such highly
sensitive spectrophotometry, small signal variation is
required.
[0004] However, there is a problem of inability of correct
measurement. Because a photodetector used for such high sensitive
spectrophotometry is so sensitive, the photodetector detects even
weak signals irrelevant to the objective measurement. As a method
for preventing such measurement errors caused by weak signals
irrelevant to the objective measurement, for example, a method for
avoiding the influence of incorrect electric current by grounding
of the photoelectric converter, or a method for detecting only
objective signals by subtracting incorrect current measured in
advance has been employed.
[0005] However, even though these procedures are performed,
problems which are disturbed the high-accuracy measurement of the
objective component remains, since background value varies by each
measurement. Therefore, correct values cannot be obtained unless a
calibration curve showing the relationship between the signal
values and the concentrations of the objective component obtained
by measuring the signal value using standard of the objective
component containing known concentrations as a sample is
established for each measurement.
[0006] For example, when a trace component is determined by using a
luminescence property of luminol, luminol is oxidized by an action
of appropriate amount of an oxidase such as peroxidase (POD) in
accordance with the amount of the objective component to be
measured in the presence of hydrogen peroxide, and fluorescence is
generated (emit light). However, even when the concentration of the
objective component is zero and the oxidase such as POD is not
present, luminol is oxidized and generates luminescence slightly.
There are problems which these energy variation value (background
value) of luminol caused by factors other than the oxidase such as
POD, or energy variation value generated by the oxidase such as POD
(they are often referred to collectively as "signal value"), are
varied significantly by change in measurement environment such as
day and time of measurement, measurement place, and the like.
[0007] Such phenomena as mentioned above are generally observed in
a method for measuring a signal value of energy variation of a
substance such as luminescence, fluorescence, phosphorescence, and
thus posing problems.
[0008] Therefore, development of a measuring method which provides
low variation of the signal values and an instrument used for the
method has been desired.
DISCLOSURE OF THE INVENTION
Problem to be Solved by the Invention
[0009] The present invention has been accomplished in view of such
circumstances as above, and an object of the present invention is
to provide a method for measuring an objective component to be
measured in high accuracy, reproducibility and simplicity, wherein
the measurement of an objective component in a sample using a
spectrophotometer is performed by suppressing or reducing a
influence of electric charge in an atmosphere surrounding a
reaction vessel which is occurred in measuring environment, and to
provide an instrument used for the method.
Means for Solving Problems
[0010] The present invention was accomplished to solve the
above-described problems, and consists of the following;
(1) A method for measuring an amount of an objective component to
be measured in a sample, which comprises;
[0011] preventing an electric charge in an atmosphere in a
photometry chamber from transferring to the surface of a solution
which generates light due to an energy variation of a substance
induced by the objective component in the sample,
[0012] measuring value of the light, and
[0013] determining an amount of the objective component in the
sample on the basis of the measured value thus obtained.
(2) A method for measuring an amount of an objective component to
be measured in a sample, which comprises; making an atmosphere
surrounding a reaction vessel and/or a reaction vessel in a
photometry chamber electrically constant, measuring value of light
generated due to an energy variation of a substance induced by the
objective component in the sample in the reaction vessel in the
photometry chamber, and determining an amount of the objective
component in the sample on the basis of the measured value thus
obtained. (3) An instrument for measuring value of light generated
due to an energy variation of a substance in a solution instrument,
which contains a mechanism for measuring value of light generated
due to an energy variation of a substance in a solution and a
mechanism for preventing an electric charge in an atmosphere in a
photometry chamber from transferring to the surface of the
solution. (4) An instrument for measuring value of light generated
due to an energy variation of a substance, which contains a
mechanism for measuring value of light generated due to an energy
variation of a substance and a mechanism for treating for making an
atmosphere surrounding a reaction vessel and/or a reaction vessel
of a photometry chamber electrically constant.
[0014] Namely, the present inventors have studied extensively in
order to find out a method for measuring an objective component to
be measured such as a trace component to be measured in high
accuracy, by suppressing or avoiding influences of variation of the
signal value in spectrophotoscopic measurement using a
spectrophotometer, etc. As the result, the present inventors have
found that the variation of the signal value can be suppressed by
preventing an electric charge in an atmosphere in a photometry
chamber from transferring to the surface of the solution which
generates light due to the energy variation of a substance induced
by the objective component in the sample, and the present invention
has been accomplished.
[0015] A measuring method of the present invention is a method for
measuring an amount of an objective component to be measured in a
sample, which comprises; preventing an electric charge in an
atmosphere in a photometry chamber from transferring to the surface
of a solution which generates light due to an energy variation of a
substance induced by the objective component in the sample,
measuring value of the light, and determining an amount of the
objective component in the sample on the basis of the measured
value thus obtained.
[0016] In the measuring method of the present invention, the method
for preventing an electric charge in an atmosphere in a photometry
chamber from transferring to the surface of a solution which
generates light due to the energy variation of a substance induced
by the objective component in the sample includes, for example, a
method for making an atmosphere surrounding a reaction vessel
and/or an atmosphere surrounding the reaction vessel in the
photometry chamber electrically constant, or a method for blocking
a contact of the atmosphere in the photometry chamber with the
solution in the reaction vessel.
[0017] The atmosphere surrounding a reaction vessel and/or the
atmosphere surrounding a reaction vessel in a photometry chamber is
defined as air with which the reaction vessel in the photometry
chamber contacts, or air in the photometry chamber equipped in the
spectrophotometer.
[0018] Light generated by the energy variation of the substance
induced by the objective component in the sample of the present
invention, includes luminescence, fluorescence or
phosphorescence.
[0019] In the present invention, the spectrophotometry denotes to
measure light intensity distribution in spectrum of luminescence,
fluorescence, phosphorescence, and the like. The photodetector
includes for example, but not limited to, a photodiode, a
photoelectron multiplier and a CCD camera. An output method
includes, for example, but not limited to, a photocounting method,
a current-output method and a voltage-output method.
[0020] The spectrophotometer used in the present invention is not
specifically limited and includes all of those generally used in
this field such as one equipped with only a spectroscopic function;
one equipped with both reagent dispensing function and the
spectroscopic function; or a automated analyzer having a reagent
dispensing function, a B/F separating function and the
spectroscopic function all together. Specific examples of such a
spectrophotometer includes Safire.TM. (Tecan Co., Ltd.) for having
only the spectroscopic function; LB953 (Berthold Technologies Co.,
Ltd.) for having both the reagent dispensing function and the
spectroscopic function; an Automated Chemiluminescence Enzyme
Immunoassay Analyzer such as SphereLight 180 (Olympus Co., Ltd.)
for the automated analyzer.
[0021] The photometry chamber in the present invention denotes a
space equipped with the photodetector in the spectrophotometer
where a sample (the reaction vessel) is set when light is actually
measured.
[0022] As the reaction vessel in the present invention, various
kinds of vessels can be used, and it is not particularly limited as
long as it is applicable to the spectrophotometer to be used, and
includes, for example, one made from glass or plastic such as
polypropylene or polystyrene. A vessel exclusive for the
spectrophotometer can also be used. In addition, there is a
cartridge type vessel for the automated analyzer having plural
concave portions (referred to as a reaction bath or a reagent cell)
in which a solution will be dispensed. This type of vessel can also
be used in the present invention.
[0023] In the present invention, "to make the atmosphere
surrounding the reaction vessel and/or the atmosphere surrounding
the reaction vessel in the photometry chamber electrically
constant" means that either the example in which electric charge of
the electrical environment in the atmosphere surrounding the
reaction vessel and/or the atmosphere surrounding the reaction
vessel in the photometry chamber is neutralized; the bias of
electric charge of the electrical environment is suppressed; or an
electric charge of the electrical environment is made constant. The
treatment is performed at just before and/or at spectrometric
measurement time, through spectrometric measurement time, or
through from just before the measurement time to the measurement
time.
[0024] The method for making the atmosphere surrounding the
reaction vessel and/or the atmosphere surrounding the reaction
vessel in the photometry chamber electrically constant is not
particularly limited, as long as it can achieve this purpose, and
includes, for example, a method for treating the atmosphere
surrounding the reaction vessel and/or the atmosphere surrounding
the reaction vessel in the photometry chamber by using one or more
of material selected from the group consisting of; (1) gas having a
constant electric charge, (2) an neutralization apparatus, or (3) a
material having a static electricity elimination effect.
[0025] Gas having a constant electric charge in the present
invention are not particularly limited as long as they are gas
having no electric charge or having a constant electric charge by
itself and no adverse influence on the spectrophotometry, and
includes, for example, commercially available cylinder contained
air, or an inert gas such as nitrogen, argon or helium. Also, air
from outside environment surrounding the measuring instrument or
other gas supplied by using publicly known equipment such as a
compressor or a pump; and ionized gas supplied, for example, by an
ionizer also. From the viewpoint of safety, hygiene and efficiency
of gas exchange in the reaction vessel, gas such as air, nitrogen
or argon are preferable. Further, from the aspects of easiness of
use and economy, to use air in outside environment surrounding the
spectrophotometer is preferable.
[0026] An equipment for supplying the gas are not particularly
limited as long as they can supply the gas to an objective site.
This includes publicly known equipment such as an ionizer, a
high-pressure steel cylinder, a compressor or a pump, and an air
pipe to be connected with the equipment for introducing the gas
toward the objective site.
[0027] Preferred examples of the ionizer are not particularly
limited as long as they are equipments providing, for example, a
unit of a discharge electrode which generates a positive or a
negative ion, and a unit of a discharge tube which releases a
positive or a negative ion. For example, commercially available
equipments such as an ionizer or an ion blower can be utilized.
[0028] When a case that a gas is supplied by the pump in the
present invention, for example, a piston pump can be used.
[0029] The neutralization apparatus in the present invention may be
a static charge eliminator which can neutralize an electric charge
from the atmosphere surrounding the reaction vessel and/or the
atmosphere surrounding the reaction vessel in the photometry
chamber by generating, for example, both positive and negative ions
from electric discharge part. And the neutralization apparatus may
have acceptable size to be installed in a spectrophotometer. For
example, a superimposed voltage type neutralization apparatus such
as an alternating-current neutralization apparatus, and a
self-discharge type neutralization apparatus which discharges
electricity by concentrating electric field at an electrode tip of
the neutralization apparatus according to electric potential of a
charged body are also included. In addition, the neutralization
apparatus may be any one of a direct-current type or an
alternating-current type, and a bar type or a blow type. For
example, a commercially available static charge eliminator (e.g.,
SJ010 SJ020, Keyence Corporation) may be used.
[0030] As the material having a static electricity elimination
effect in the present invention, a sheet having a static
electricity elimination effect (hereinafter abbreviate as "an
anti-static sheet") such as a tape having a static electricity
elimination effect (hereinafter abbreviate as "an anti-static
tape") (e.g. "tape anti-static" Japan Vilene Company) may be
applied. For example, a material which contains an electric
conducting layer and its supporter as a constituent, and more
specifically the material described in JP-A 5-174991, JP-A
7-243174, JP-A 5-263318 and JP-A 6-215854 are included.
[0031] In more detail, the supporter of the electric conducting
layer includes, entangled nonwoven fabrics, fabrics or knits
composed of natural fibers such as silk, wool, cotton, flax;
regenerated fibers such as rayon fiber, semi-synthetic fibers such
as acetate fiber, synthetic fibers containing one component such as
polyamide fiber, polyvinyl alcohol fiber, acrylic fiber, polyester
fiber, polyvinyliden chloride fiber, polyvinyl chloride fiber,
polyurethane fiber, polyethylene fiber, polypropylene fiber; and
composite fibers of the core-in-shell type, or the side by side
type containing two or more components.
[0032] A component of the electric conducting layer includes
polymers produced by polymerization of a monomer such as acetylene,
benzene, aniline, phenylacetylene, pyrrole, furan, thiophene,
indole and their derivatives. These polymers may be processed with
an electrically conductive substance (for example, a metal such as
stainless steel, aluminum, zinc, cupper, tin and silver, or carbon)
before or after coating over the above-mentioned supporter. The
above-mentioned supporter is coated almost totally with these
polymers to form the electric conducting layer.
[0033] Further, the anti-static sheet preferably has a
non-conducting layer. For example, the sheet having an adhesive
layer on the supporter so that it is attached at the right position
for carrying out an elimination of static electricity is
preferable. Typically, the adhesive layer composed of, for example,
thermoplastic synthetic resins such as homopolymers or copolymers
derived from such a monomer as an acrylic ester, vinyl acetate,
vinyl chloride, vinylidene chloride, an acrylic monomer, styrene
and an olefin; rubber such as natural rubber, natural-synthetic
rubber, nitrile rubber, styrene-butadiene rubber, chloroprene
rubber, butyl rubber, or thermosetting resins such as an
urea-formalin resin, a melanin-formalin resin and a phenolic resin
may be formed.
[0034] More typically, the sheet having a static electricity
elimination effect includes, for example, but not limited to, a
anti-static sheet having an entangled nonwoven fabric which is
coated almost as a whole with an electron conjugated polymer, and
that is formed an electric conducting layer having a higher
electric conductivity than the electron conjugated polymer (JP-A
5-174991), an ant-static sheet composed of a fiber sheet of
synthetic rubber and/or elastomer as a main component in which the
surface is coated with an electron conjugated polymer (JP-A
7-243174), and a cloth having a static electricity elimination
effect which woven with conductive composite fibers composed of a
protective polymer layer, a covering polymer layer and an electric
conducting layer (JP-A 5-263318). In addition, sheet substances
formed with conductive fibers such as metal plated conductive
fiber, metal fiber, carbon fiber, metallic ion containing fiber,
and sheet substance obtained by fiber sheet formulation using such
as regenerated fiber, semi-synthetic fiber, synthetic fiber,
inorganic fiber, plant fiber, animal fiber and mineral fiber,
followed by furnishing conductivity (JP-A 6-215854).
[0035] In the present invention, the above-mentioned methods (1),
(2) and (3) may be used alone or in arbitrary combination.
[0036] More detailed explanation for each procedure is given
below.
(1) A Method for Treating with Gas having a Constant Electric
Charge
[0037] A typical example of the method includes, for example, (i) a
method for blowing more than a certain amount of gas having a
constant electric charge to the reaction vessel, (ii) a method for
exchanging the atmosphere in the photometry chamber with gas having
a constant electric charge by circulating gas having a constant
electric charge.
[0038] As to the method (i), when an automated analyzer is
utilized, for example, a gas circulation pipe led from an equipment
for supplying the gas having a constant electric charge (e.g. a
pump, or an ionizer) is installed in the former position of the
photometry chamber to where the reaction vessel of the analyzer is
moved, and the gas may be blown to the reaction vessel at a stage
before the reaction vessel is moved to the photometry chamber.
[0039] Further, such procedure may be set up as to blow gas having
a constant electric charge to the reaction vessel every time when
the reaction vessel passes through in front of the gas circulation
pipe. When the automatic analyzer is set to allow the gas to
circulate in such area continuously during spectrophotometric
measurement, the above-mentioned set up of blowing schedule can be
eliminated.
[0040] When a sample prepared by manual means is subjected to the
measurement using a spectrophotometer, the reaction vessel may be
blown by gas having a constant electric charge, and then the
reaction vessel may be set to the photometric area.
[0041] As to the method (ii), for example, when an automated
analyzer is utilized, the method described below is included as an
example. That is, a gas circulation pipe led from an equipment for
supplying the gas having a constant electric charge (e.g. a pump,
or an ionizer) is installed in the automated analyzer, and makes
the gas flow in the photometry chamber. In this regard, since the
photometry chamber has to be light shielded, the gas circulation
pipe is set in former position of the photometry chamber rather
than in the photometry chamber, and allows the gas flow toward the
photometry chamber during the measurement. And when the reaction
vessel in which a sample and reagents are held is moved to the
photometry chamber and a shielding board is opened, the gas is
supplied to the photometry chamber through the gas circulation
pipe, followed by closing the shielding board, and thus light
shielding of the photometry chamber may be maintained. So, this
method is preferable.
[0042] Alternatively, gas may be flowed in every time when the
shielding board opens. When the gas is kept flowing continuously
during the photometric measurement, it is not necessary to rely on
such complicated set up as gas flowing at every opening time of the
shielding board.
[0043] When a sample prepared by manual means is subjected to the
measurement using a spectrophotometer, the gas circulation pipe led
from an equipment for supplying the gas having a constant electric
charge is set up to the around of the photometry chamber, similarly
as in an automated analyzer, and the gas may be blown to the
photometric area through the gas pipe.
[0044] In case where the method (i) is enforced, the amount and
strength (flow rate) of the gas having a constant electric charge
to be blown to the reaction vessel may be such level as sufficient
to remove an electric charge of the reaction vessel. In case where
the method (ii) is enforced, the amount and flow rate of gas having
a constant electric charge to be circulated to the photometric area
may be set to be able to introduce the sufficient amount of gas to
exchange gas in the objective photometry chamber by the gas.
(2) Method for Using a Neutralization Apparatus
[0045] As to a typical example of the method, when an automated
analyzer is utilized, for example, a neutralization apparatus is
installed in just former position to where the reaction vessel of
the analyzer is moved into the photometry chamber, and the reaction
vessel can be passed through the effective area of the
neutralization apparatus (charge neutralization area) at a stage
before the reaction vessel is moved into the photometry
chamber.
[0046] When the above method is enforced, such a procedure may be
adopted as the reaction vessel is subjected to ion exposure every
time when each reaction vessel set on a turning table passes in
front of the ion-discharging site of the neutralization apparatus.
When the neutralization apparatus is set to work throughout the
spectrophotometric measurement to allow the reaction vessels come
serially into the charge neutralization area one after another, the
above-mentioned set up for charge neutralization program is not
required.
[0047] For enforcement of the method, the amount of ion exposure to
the reaction vessel may be required at such level as sufficient to
neutralize electric charge of the reaction vessel.
(3) A Method for Using Material Having a Static Electricity
Elimination Effect
[0048] As for the method, there is, for example, a method for
attaching a material having a static electricity elimination effect
to an inside wall surface of a photometric camber (e.g. an
anti-static sheet). The anti-static sheet may be attached to any
place in the photometry chamber unless the measurement operation is
disturbed. Also, the size of the anti-static sheet is not
especially limited, as long as it is not too large without
additional effect and uneconomical. In addition, it is not
necessary to attach the sheet all-around the inside wall surface of
the photometry chamber. When the volume of the photometry chamber
is about 4 cm.times.3 cm.times.8 cm, belt-like attachment on the
inside wall surface by a tape-shaped one having 2 to 3 cm width may
be sufficient.
[0049] In addition, in the case of the reaction vessel is a
cartridge type, the ant-static sheet may be attached along the
outside of the cartridge. The anti-static sheet may also be
attached both along the outside of the cartridge and inside wall
surface of the photometry chamber. In particular, to attach the
anti-static sheet at the position where the cartridge is sandwiched
between both sides is effective. Typically, this includes, for
example, (i) a method for attaching an anti-static sheet on the
inside wall surface of the photometry chamber parallel to the
longitudinal side surface of the cartridge, (ii) a method for
attaching an anti-static sheet on the longitudinal outside surface
of the cartridge, and (iii) a method for carrying out both methods
(i) and (ii).
[0050] Among the above three treatment methods (1) to (3), the
method (3) of using material having a static electricity
elimination effect is the simplest procedure.
[0051] Also, the method for blowing gas having a constant electric
charge to the reaction vessel at the stage before the reaction
vessel is moved to the photometry chamber, or the method for using
the neutralization apparatus is also simple and preferable
procedure.
[0052] In the present invention, as "a method for blocking a
contact of the atmosphere in the photometry chamber with the
solution in the reaction vessel" includes, for example, (1) a
method for shutting an opening part of the reaction vessel holding
the solution which generates light due to the energy variation of
the substance induced by the objective component in the sample with
a sheet, or (2) a method for covering the surface of the solution
in the reaction vessel with a substance insoluble to the
solution.
[0053] In the method (1) for shutting the opening part of the
reaction vessel holding the solution with a sheet includes, for
example, a method for shutting the opening part of the reaction
vessel holding the solution with a sheet such as a cellophane tape,
an aluminum seal, an aluminum foil, a paper tape or a plastic tape
and the like which can block a contact of the atmosphere in the
photometry chamber with the solution in the reaction vessel. In so
doing, it is preferable to shut whole of the opening parts by
sealing to prevent leaking of the atmosphere in the reaction vessel
to outside of it, but complete seal is not essential.
[0054] Preferable embodiments of the reaction vessel in which the
solution is held are described above.
[0055] The method (2) for covering the surface of the solution
which generates light due to energy variation of the substance
induced by the objective component in the sample with a substance
insoluble in the solution in the reaction vessel includes, (i) a
method for blocking the contact of the atmosphere in the photometry
chamber with the solution in the reaction vessel by covering the
surface of the solution with liquid which is insoluble in the
solution in the reaction vessel, and the liquid has a property of
spreading over the surface of the solution in the vessel, and (ii)
a method for blocking a contact of the atmosphere in the photometry
chamber with the solution in the reaction vessel by covering the
surface of the solution by applying (floating) a substance which
has lower specific gravity than the solution in the reaction vessel
and is insoluble in the solution and has a property of floating on
the surface of the solution.
[0056] The liquid which is insoluble in the solution in the
reaction vessel, and having the property of spreading over the
surface of the solution in the vessel used in the method (i)
includes, for example,
silicon oil, hexane and liquid paraffin, etc. when the solution in
the reaction vessel is an aqueous solution, but not limited
thereto. When the solution in the reaction vessel is non-aqueous
solution, the liquid includes, for example, hexane and benzene, but
not limited thereto.
[0057] The substance which has lower specific gravity than the
solution in the reaction vessel and is insoluble in the solution
and has a property of floating on the surface of the solution used
in the method (ii) includes, for example, cork and polystyrene
foam, but not limited thereto.
[0058] The method for measuring an amount of the objective
component in the sample of the present invention may be performed
by measuring the energy variation of a substance induced by the
presence of the objective component in the sample, for example,
luminescence, fluorescence or phosphorescence using a known
spectrophotometer and measurement reagents, and according to known
measurement conditions (e.g., reaction time, measured wave length,
etc.), and a procedure for performing an ELISA, etc., except for
treating the atmosphere surrounding the reaction vessel and/or the
atmosphere surrounding the reaction vessel in the photometry
chamber, or blocking a contact of the atmosphere in the photometry
chamber with the solution in the reaction vessel, and are not
particularly limited. For details, the measurement may be performed
by using each measuring instrument and reagents, according to a
manual of each instruments and reagents.
[0059] A practical measurement method for generating an energy
variation of a substance induced by the objective component in the
sample, is not particularly limited, as long as it is a method for
making the above-described substance which generates light present
in the solution together with the sample containing the objective
component to be measured, carrying out various types of reaction
which generates energy variation in the presence of the substance,
and measuring the amount of light such as luminescence,
fluorescence or phosphorescence resulted from the reaction, and
measuring the objective component in the sample using the measured
value thus obtained.
[0060] This kind of reaction includes the known method, for
example, the method for reacting the objective component with an
oxidase of which the objective component is a substrate to produce
hydrogen peroxide in a solution such as an appropriate buffer
solution, and generating an appropriate energy variation by
reacting this hydrogen peroxide with an appropriate chemical
compound, and the method for reacting an labeled antibody
specifically bind to the objective component which labeled with an
oxidase such as POD with the objective component to produce an
immune complex of the objective component and the labeled antibody,
and then reacting the (POD labeled) immune complex with a substrate
such as hydrogen peroxide and luminol, and measuring the generated
energy variation (for example, luminescence, fluorescence or
phosphorescence) by the reaction.
[0061] Besides, in the above-described method, the objective
component may participate in main reaction either directly or
secondarily (indirectly).
[0062] In order to avoid influences of substances other than the
objective component contained in the sample, or to increase the
detection sensitivity to the objective component, additives
commonly used in this field such as a surfactant or an activator
agent may be added.
[0063] Such measurement method includes those generally used, for
example, a chemical measurement method (e.g. chelation luminescence
method), an enzymatic measurement method (e.g. an
oxidation-reduction luminescence method), and an immunological
measurement method [e.g. an enzyme immunoassay (ETA), a
fluoroimmunoassay (FIA), an immunological inhibition method, a
chemiluminescence method, etc.] used in the field of, for example,
clinical laboratory, biochemistry, biology, chemistry, food and
environmental research. The measurement mechanism thereof may be a
variety of procedures including a sandwich method, a competitive
method and a double antibody technique, etc.
[0064] A chemiluminescent substance, a fluorescent substance or
phosphorescent substance in order to measure luminescence,
fluorescence or phosphorescence induced by the presence of the
objective component to be measured in a measurement method for the
present invention includes a substance usually used in the fields
such as enzyme immunoassay (ETA), radioimmunoassay (RIA) and
fluoroimmunoassay (FIA).
[0065] The chemiluminescent substance includes, for example,
luminol derivatives such as isoluminol, luminol,
aminoethylisoluminol, aminoethyl ethyl isoluminol,
aminopropylisoluminol, aminobutylisoluminol, aminohexyl ethyl
isoluminol, etc.; and luciferin, bis (2,4,6-trifluorophenyl)
oxalate, etc.
[0066] The fluorescent substance includes, for example, a substance
usually used in fluorescent immunoassay such as fluorescein,
dansyl, fluorescamine, coumarin, naphthylamine, fluorescein
isothiocyanate, rhodamine, rhodamine-X, sulforhodamine 101, lucifer
yellow, acridine, acridine isothiocyanate, riboflavin or
derivatives thereof and europium (Eu), etc.
[0067] The phosphorescent substance includes, for example, a
mixture of sulfides of alkaline earth metals (e.g., calcium, and
the like) and a small amount of heavy metals (e.g., cupper, and the
like), methyl substituted benzoxazole, or benzophenone.
[0068] The measurement may be performed by generating luminescence
by a luminescent substance, fluorescence by a fluorescent substance
and phosphorescence by phosphorescent substance, corresponding to
the amount of the objective component in the sample, and measuring
the respective signal value.
[0069] For example, the method for measuring TSH
(thyroid-stimulating hormone) in a sample by the method of the
present invention is exemplified below.
[0070] A sample, polystyrene beads immobilized with an anti-TSH
monoclonal antibody, POD-labeled anti-TSH monoclonal antibody
solution, a 5 mM luminol solution and a H.sub.2O.sub.2 solution are
each loaded on an automated analyzer which equipped with a
mechanism for making the atmosphere surrounding the reaction vessel
and/or the atmosphere surrounding the reaction vessel in the
photometry chamber electrically constant of the present invention.
After that, the analyzer is activated, the sample containing TSH
and polystyrene beads immobilized with the anti-TSH monoclonal
antibody are added and reacted, followed by B/F separation,
POD-labeled anti-TSH monoclonal antibody is added thereto and
reacted. Subsequently, the beads after B/F separation are added
with a 5 mM luminol solution (a luminescent substrate) and a
H.sub.2O.sub.2 solution, mixed, and then luminescence of the sample
may be measured.
[0071] When a common spectrophotometer is used instead of the
automated analyzer for measuring the above-described TSH, a sample
and various reagents are reacted by manual means, luminescent
reaction is occurred, after that, the reaction vessel is set in a
spectrophotometer equipped with the mechanism for making an
atmosphere surrounding the reaction vessel and/or the atmosphere
surrounding the reaction vessel in the photometry chamber
electrically constant of the present invention, and then,
luminescence of the sample may be measured.
[0072] Additionally, another embodiment of the measuring method of
the present invention includes, luminescence in a sample may be
measured by using the spectrophotometer equipped with a mechanism
for blocking the contact of the atmosphere in the photometry
chamber with the solution in the reaction vessel. Typically, for
example, the methods (i), (ii) or (iii) described below are
included.
[0073] (i) A sample, polystyrene beads immobilized with an anti-TSH
monoclonal antibody, peroxidase (POD)-labeled anti-TSH monoclonal
antibody solution, a 5 mM luminol solution and a H.sub.2O.sub.2
solution are each loaded on an automated analyzer which equipped
with a mechanism for shutting the opening part of the reaction
vessel with the sheet of the present invention. After that, the
analyzer is activated, the sample containing TSH and polystyrene
beads immobilized with the anti-TSH monoclonal antibody are added
and reacted, followed by B/F separation, peroxidase (POD)-labeled
anti-TSH monoclonal antibody is added thereto and reacted. After
that, the beads after B/F separation are added with a 5 mM luminol
solution (a luminescent substrate) and a H.sub.2O.sub.2 solution,
and mixed. Subsequently, the opening part of the reaction vessel is
shut with the sheet by a mechanism equipped in the automated
analyzer, and then luminescence of the sample may be measured.
[0074] When a common spectrophotometer is used instead of an
automated analyzer for measuring the above-described TSH, a sample
and various reagents are reacted by a manual means, luminescent
reaction is occurred, after that, the opening part of the reaction
vessel is shut by sealing, for example, with a cellophane tape, and
then, the reaction vessel is set in a spectrophotometer, and then,
luminescence of the sample may be measured.
[0075] (ii) A sample, polystyrene beads immobilized with an
anti-TSH monoclonal antibody, peroxidase (POD)-labeled anti-TSH
monoclonal antibody solution, a 5 mM luminol solution and a
H.sub.2O.sub.2 solution are each loaded on an automated analyzer
which equipped with a mechanism for dropping on (or run in) liquid
which is insoluble in the solution in the vessel and has a property
of spreading over the surface of the solution in the vessel. After
that, the analyzer is activated, the sample containing TSH and
polystyrene beads immobilized with an anti-TSH monoclonal antibody
are added and reacted, followed by B/F separation, peroxidase
(POD)-labeled anti-TSH monoclonal antibody is added thereto and
reacted. After that, the beads after B/F separation are added with
a 5 mM luminol solution (a luminescent substrate) and a
H.sub.2O.sub.2 solution, and mixed. Subsequently, liquid which is
insoluble in the solution in the vessel and has a property of
spreading over the surface of the solution is dropped on (or run
in) the surface of the solution in the reaction vessel to make a
water-insoluble membrane by using a mechanism equipped in the
automated analyzer, and then luminescence of the sample may be
measured.
[0076] When a common spectrophotometer is used instead of an
automated analyzer for measuring the above-described TSH, a sample
and various reagents are reacted by a manual means, luminescent
reaction is occurred, after that, liquid which is insoluble in the
solution in the vessel and has a property of spreading over the
surface of the solution in the reaction vessel is dropped or run in
gently alongside the wall of a reaction vessel on the surface of
the solution to make a water-insoluble membrane, and the reaction
vessel is set in a spectrophotometer, and then, luminescence of the
sample may be measured.
[0077] (iii) A sample, polystyrene beads immobilized with an
anti-TSH monoclonal antibody, peroxidase (POD)-labeled anti-TSH
monoclonal antibody solution, a 5 mM luminol solution and a
H.sub.2O.sub.2 solution are each loaded on an automated analyzer,
which equipped with a mechanism for applying (floating) a substance
which has lower specific gravity than the solution in the reaction
vessel, insoluble in the solution and has a property of floating on
the surface of the solution on the surface of the solution. After
that, the analyzer is activated, the sample containing TSH and
polystyrene beads immobilized with the anti-TSH monoclonal antibody
are added and reacted, followed by B/F separation, peroxidase
(POD)-labeled anti-TSH monoclonal antibody is added thereto and
reacted. After that, the beads after B/F separation are added with
a 5 mM luminol solution (a luminescent substrate) and a
H.sub.2O.sub.2 solution, and mixed. Subsequently, the substance
which is insoluble in the solution in the vessel and has lower
specific gravity than the solution and has a property of floating
on the surface of the solution is applied (floated) to cover the
surface of the solution in the vessel by using a mechanism equipped
in the automated analyzer, and then luminescence of the sample is
measured.
[0078] When a common spectrophotometer is used instead of an
automated analyzer for measuring the above-described TSH, a sample
and various reagents are reacted by a manual means, luminescence
reaction is occurred, after that, the substance which is insoluble
in the solution in the reaction vessel and has lower specific
gravity than the solution in the vessel and has a property of
floating on the surface of the solution is applied (floated) to
cover the surface of the solution in the vessel, and the reaction
vessel is set in a spectrophotometer, and then, luminescence of the
sample is measured.
[0079] A method for obtaining an amount of the objective component
in the sample on the basis of the measured values thus obtained may
be determined in accordance with a protocol of each
spectrophotometer used. For example, the signal value obtained by
the above-mentioned method is applied to a calibration curve
showing a relationship between concentration and a signal value
which is previously obtained by measuring a standard solution
containing a known amount of the objective component after a
similar method as mentioned above whereby an amount of the
objective component in the sample can be obtained. In addition, in
the method described above, an amount of the objective component in
the sample can be determined by subtracting a blind value, which is
obtained by measuring a sample in which an objective component is
not contained after a similar method as mentioned above from the
measured signal value.
[0080] The objective components which can be measured by the method
of the present invention are not particularly limited and any
component can be measured as long as it has reactivity to produce
luminescence, fluorescence, phosphorescence or absorption of light
corresponding to the amount of the objective component to be
measured [an objective component may participate in main reaction
of the reaction either directly or secondarily (indirectly)], and
which can be measured by spectrophotometry according to the
generated light. Such component includes, for example,
environmental hormone such as estrogen and bisphenol-A; tumor
markers such as .alpha.-fetoprotein (AFP), CA19-9, a
prostate-specific antigen (PSA) and a carcinoembryonic antigen
(CEA); serum proteins such as immunoglobulin A (IgA),
immunoglobulin E (IgE), immunoglobulin G (IgG), immunoglobulin M
(IgM), .beta..sub.2-microglobulin and albumin; enzymes such as
amylase and alkaline phosphatase; in vivo substances such as
cholesterol, triglyceride, creatinine and uric acid; drugs such as
steroids, acetaminophens and digoxin compounds; non-peptide hormone
such as estradiol and prolactins; DNA measuring reagents such as
oligonucleotides complemental to nucleic acid components derived
from various microorganisms, mycoplasmas and various viruses.
[0081] Examples of a sample are those containing the objective
component to be measured as described above, and includes, for
example, a sample derived from a living body which is exemplified
by various body fluids such as serum, plasma, spinal fluid, saliva
and pancreatic fluid, excrements such as urine and fecal matter (or
its dilution), lymphocytes, blood cells, various cells, and tissue
extracts of living body; a sample derived from plants such as plant
tissue extracts or cell extracts; a sample derived from
microorganisms such as culture medium and cell extracts; and a
sample derived from food derivatives such as food staff or extracts
thereof. These samples are usually used in the field of clinical
laboratory, biochemistry, biology, chemistry of food science,
etc.
[0082] The instrument of the present invention contains a mechanism
for measuring value of light generated due to the energy variation
of a substance and (a) a mechanism for making an atmosphere
surrounding a reaction vessel and/or an atmosphere surrounding the
reaction vessel in the photometry chamber electrically constant, or
(b) a mechanism for blocking a contact of an atmosphere in the
photometry chamber with the solution in a reaction vessel.
[0083] A typical examples of the mechanism in the above (a) for
making an atmosphere surrounding a reaction vessel and/or an
atmosphere surrounding the reaction vessel in the photometry
chamber electrically constant includes one or more of mechanisms
selected from the group consisting of; (1) an apparatus for flowing
gas having a constant electric charge to surrounding the reaction
vessel and/or into the photometry chamber, (2) an neutralization
apparatus, or (3) a photometry chamber on which a material having a
static electricity elimination effect is attached. Such mechanism
is not limited to the above concrete examples as long as it can
make the atmosphere surrounding the reaction vessel and/or the
atmosphere surrounding the reaction vessel in the photometry
chamber electrically constant according to the present invention.
The typical examples of each constitutional element are as
described above. That is, the instrument of the present invention
can be obtained by equipping one or more of mechanism selected from
examples described above, to a spectrophotometer used for the
present invention.
[0084] A typical examples of mechanism (b) for blocking a contact
of the atmosphere in the photometry chamber with the solution in
the reaction vessel includes one or more of mechanism selected from
the group consisting of; (1) an mechanism for shutting an opening
part of a reaction vessel holding the solution with a sheet, (2) a
mechanism for covering the surface of the solution with liquid
which is insoluble in the solution and spreads over surface of the
solution by dropping (or running in), (3) a mechanism for applying
(floating) a substance which has lower specific gravity than the
solution in the reaction vessel, insoluble in the solution and has
a property of floating on the surface of the solution. Such
mechanism is not limited to the above concrete examples as long as
it can block a contact of the atmosphere in the photometry chamber
with the solution in the reaction vessel according to the present
invention. The typical examples of each constitutional element are
as described above.
[0085] In addition, an instrument of the present invention may be
equipped either with the above-described (a) the mechanism for
making the atmosphere surrounding the reaction vessel and/or the
atmosphere surrounding the reaction vessel in the photometry
chamber electrically constant, or (b) the mechanism for blocking a
contact of the atmosphere in the photometry chamber with the
solution in the reaction vessel either independently or in
combination of (a) and (b). Quite naturally, such mechanism may
comprise both of plural mechanism (a) and plural mechanism (b) both
in plural number.
[0086] Therefore, according to the present invention, the
instrument of the present invention can be obtained only by
equipping the above-described mechanism to the spectrophotometer
which is conventionally used. This provides great advantage due to
no need for measuring instrument equipped with a particular kind of
mechanism for preventing the influence of the electric charge.
[0087] According to the present invention, between-day variation of
signal value and background value can be reduced easily and simply
only by making an atmosphere surrounding the reaction vessel and/or
the atmosphere surrounding the reaction vessel in the photometry
chamber electrically constant, or by blocking a contact of the
atmosphere in the photometry chamber with the solution in the
reaction vessel. In other words, a conventional problem was that
accurate measurement could not be performed because of variation of
signal value when a in vivo trace component is determined by
measuring luminescence, fluorescence, phosphorescence and the like.
After all, a problem lasted for a long time without favorable
improvement, even though various intensive studies have been
carried out to overcome such trouble. These problems can be solved
easily and quite unexpectedly by the present invention.
EFFECTS OF THE INVENTION
[0088] According to the present invention, in measurement of an
objective component to be measured in a sample by
spectrophotometric measurement of light generated due to an energy
variation using a spectrophotometer, problems such as between-day
variation of signal value or increase of background value can be
solved, and a trace component can be measured in high accuracy and
high sensitivity.
[0089] The present invention will be explained further in detail by
referring to Examples, but the present invention is not construed
as limiting to these Examples.
EXAMPLES
Experimental Example 1
Investigation of a Variation of Measured Value Caused by
Environmental Changes
[0090] [Preparation of Reagents]
[0091] Following reagents were prepared:
[0092] Distilled water,
[0093] Reagent A: a solution of 5 mM luminol (a buffer solution
exclusive for SphereLight 180, pH 8.5, Wako Pure Chemical
Industries, Ltd.),
[0094] Reagent B: an aqueous solution of 0.02% H.sub.2O.sub.2,
[0095] Reagent C: a mixture of equal volume of Reagent A and
Reagent B.
[0096] [Measurement of Luminescence]
[0097] Luminescence of 140 .mu.L of each reagent prepared in above
was measured by using SphereLight 180 which was automated
chemiluminescence enzyme immunoassay analyzer (SphereLight 180,
Olympus Ltd.) as a measuring instrument. Measurement was repeated
10 times (every 20 seconds in this Example) at the same interval in
the same sample.
[0098] Also, the measurement was carried out 10 times at different
days by the same method as described above, and the variation of
measured value (signal value) for every measurement and the
between-day variation were investigated.
[Results]
[0099] Results are shown in Table 1-1 and Table 1-2. Table 1-1
shows the results of the first measurement day, and Table 1-2 shows
results of the second measurement day (Unit: cps).
TABLE-US-00001 TABLE 1-1 Number of Measurement Water Reagent A
Reagent B Reagent C 1 19 19 11 1,199 2 11 21 10 1,118 3 21 19 21
1,174 4 16 24 20 1,167 5 9 29 19 1,118 6 20 25 17 1,144 7 9 28 10
1,134 8 14 37 14 1,174 9 18 21 12 1,168 10 10 26 15 1,188 Average
14.7 24.9 14.9 1,158.4
TABLE-US-00002 TABLE 1-2 Number of Measurement Water Reagent A
Reagent B Reagent C 1 16 86 18 2,452 2 9 107 12 2,126 3 22 78 11
2,531 4 13 80 17 2,262 5 12 76 16 2,221 6 11 79 20 2,943 7 18 90 13
2,174 8 12 86 17 3,035 9 17 87 15 2,173 10 20 79 11 2,091 Average
15.0 84.8 15.0 2,400.8 Difference 0.3 59.9 0.1 1,242.4 Compared
with Table 1-1 (cps)
[0100] As is clear from Table 1-1 and Table 1-2, any variation of
signal value is not observed in water and reagent B containing
H.sub.2O.sub.2 even when luminescence is measured at different day
(water: 0.3 cps, H.sub.2O.sub.2: 0.1 cps). On the contrary, reagent
A and reagent C both containing luminol shows extreme between-day
variation (between-day variation of reagent A: 59.5 cps,
between-day variation of reagent C: 1,242.4 cps).
[0101] Any variation of signal value is not observed in the
measurement using water as a sample. Therefore, it is suggested
that a noise and incorrect electric current running in a
current-voltage converter are not a variation factor caused by
environmental change at measured time. Also, the variation of the
signal value is observed only in the reagent containing luminol.
Therefore, it is suggested that energy condition of luminol is
changed by environmental change at measured time, and then
variation of background value is induced.
Example 1
Removal Effect of Electricity Using Anti-Static Tape
[0102] [Preparation of Reagents]
[0103] Following reagents were prepared:
[0104] A Luminescent substrate Solution: a solution of 5 mM luminol
(a buffer solution exclusive for SphereLight 180, pH 8.5, Wako Pure
Chemical Industries, Ltd.),
[0105] A H.sub.2O.sub.2 Solution: an aqueous solution of 0.02%
H.sub.2O.sub.2 (an acid buffer solution exclusive for SphereLight
180, pH 3.0, Wako Pure Chemical Industries, Ltd.).
[Measuring Instrument]
[0106] SphereLight 180 which was automated chemiluminescence enzyme
immunoassay analyzer (Olympus Ltd.) was used. One instrument
attached the anti-static tape (25 mm wide, Japan Vilene Co., Ltd.)
transversely on four inside wall surfaces of the photometry
chamber, and the other instrument without attachment of anti-static
tape were prepared.
[Measurement of Luminescence]
[0107] The luminescent substrate solution of 70 .mu.L and 70 .mu.L
of a H.sub.2O.sub.2 solution prepared as above were automatically
dispensed into the reactor tank of the reagent cartridge in
SphereLight 180, and then, the luminescence of 140 .mu.L in total
volume of the reaction solution was measured. Incidentally, there
is a concave portion to dispense the reagent solution in the
above-described reagent cartridge, and it is called as a reactor
tank (or called as a measurement vesell).
[0108] Measurement was repeated 4 times (every 20 seconds in this
Example) at the same interval in the same sample.
[Results]
[0109] Results are shown in Table 2 (Unit: cps). In Table 2,
"Control" means a result obtained by measuring luminescence using
the measuring instrument on which an anti-static tape is not
attached (Counter measure for preventing the influence of the
electric charge is not provided.). The "versus Control" (%) is
shown as a ratio of the average of signal values obtained by
measuring luminescence using measuring instrument on which the
anti-static tape is attached, to the average of the signal value of
control.
TABLE-US-00003 TABLE 2 Number of with Anti- versus Measuremnet
Control static Tape Control 1 5,726 3,125 2 5,491 3,356 3 5,275
3,289 4 5,599 3,054 Average 5,523 3,206 58%
[0110] As is clear from Table 2, the signal values measured by
using the measuring instrument attached the anti-static tape
thereon is 58% against control. Therefore, it is understood that
the measurement using anti-static tape can suppress background
value.
Example 2
Investigation of Attaching Position of an Anti-Static Tape
[0111] [Preparation of reagents] The same reagents as in Example 1
were used.
[Measuring Instrument]
[0112] In SphereLight 180 which was automated chemiluminescence
enzyme immunoassay analyzer (Olympus Ltd.), two cases of the
apparatus were set, that is the one with an anti-static tape (25 mm
wide, Japan Vilene Co., Ltd.) attached on various positions of the
inside wall surface of the photometry chamber (Patterns 2 to 8 in
Table 3) and the other one without attachment of anti-static tape
(Pattern 1 in Table 3).
[0113] Also, the attached position of the anti-static tape in the
photometry chamber in patterns 1 to 8 was indicated by boldface in
the Table 3 showing the photometry chamber and the set position of
the reagent cartridge in the photometry chamber (Patterns 3 to 5
were set up so that the total size of attached anti-static tape
were the same.).
[0114] That is, patterns 1 to 8 each show positions of the
anti-static tapes were attached as follows:
Pattern 1: The case which the anti-static tape was not attached
(Countermeasure for preventing the influence of the electric charge
was not provided. Control). Pattern 2: The case which anti-static
tapes were attached on four inside wall surfaces (face A to face D)
of the photometry chamber. Pattern 3: The case which the
anti-static tapes were attached on face A and face B of the
photometry chamber. Pattern 4: The case which the anti-static tapes
were attached on face C and face D of the photometry chamber.
Pattern 5: The case which the anti-static tape was attached on only
face A of the photometry chamber. Pattern 6: The case which the
anti-static tape was attached on only face B of the photometry
chamber. Pattern 7: The case the anti-static tapes were attached on
longitudinal both sides of the reagent cartridge parallel to face A
and face B of the photometry chamber. Pattern 8: The case which the
anti-static tapes were attached on longitudinal both sides of the
reagent cartridge parallel to face A and face B of the photometry
chamber, and also, on face A and face B of the photometry
chamber.
[0115] Furthermore, a view showing a frame format of the photometry
chamber in case of pattern 5 is shown in FIG. 1. In FIG. 1, four
inside wall surfaces of the photometry chamber were shown as A, B,
C and D each, and these symbols correspond to A, B, C and D in the
Figure showing the attached position for attachment of an
anti-static tapes in photometry chamber in pattern 1 of Table 3,
respectively. Also, the attached position of the anti-static tape,
the position of the photometry chamber, the position of the
photoelectron multiplier against to the photometry chamber, and
setting position of the reagent cartridge holding the reaction
solution are shown in FIG. 1. That is, the anti-static tape was
attached in the photometry chamber, and the photoelectron
multiplier is located on wall side of face A of a photometry
chamber.
[0116] [Measurement of Luminescence]
[0117] The luminescent substrate solution of 70 .mu.L and 70 .mu.L
of the H.sub.2 O.sub.2 solution prepared as above were
automatically dispensed into the reactor tank of the reagent
cartridge in SphereLight 180, and then, luminescence of 140 .mu.L
in total volume of reaction solution was measured. Measurement was
repeated 5 times (every 20 seconds in this Example) at the same
interval in the same reaction solution.
[Results]
[0118] Results are shown in Table 3 (Unit: cps). In Table 3, A, B,
C and D shown in Figure of attached position of anti-static tape in
photometry chamber in pattern 1 corresponds to A, B, C and D in
FIG. 1, respectively.
[0119] Also, the result (Pattern 1) obtained by measuring
luminescence using the measuring instrument on which an anti-static
tape is not attached is determined as control. Also, "versus
Control" (%) is shown as a ratio of the average of the signal
values obtained by measuring luminescence using measuring
instrument of each pattern, to the average of the signal value of
control.
TABLE-US-00004 TABLE 3 ##STR00001## ##STR00002##
[0120] As is clear from Table 3, it is understood that background
value can be suppressed when luminescence is measured by using the
instrument attached the anti-static tapes thereon.
[0121] In particular, in patterns 2, 3, 4, 7 and 8 of Table 3,
background value is found to be suppressed to not higher than 50%
(value of "versus Control" is low). From the results, it is found
that to attach the anti-static tape on the position where the
reagent cartridge is sandwiched between both sides is especially
effective.
Example 3
Influence of Electrification of a Reagent Cartridge and Grounding
Effect on the Measurement
[0122] [Preparation of Reagents] The same reagents as in Example 1
were used.
[Measuring Instrument]
[0123] In SphereLight 180 which was automated chemiluminescence
enzyme immunoassay analyzer (Olympus Ltd.), aluminum foil was set
at the undersurface of the photometry chamber (set position of
aluminum foil was shown by oblique lines in FIG. 2), and also, an
anti-static tape (25 mm wide, Japan Vilene Co., Ltd.) was attached
on four inside wall surfaces of the photometry chamber.
[0124] In addition, when the reagent cartridge is set (FIG. 1), the
reagent cartridge contacts with aluminum foil which was set at the
undersurface of the photometry chamber. And the aluminum foil
contacts with the screw at the undersurface of the photometry
chamber, and the screw is connected to the grounding of the
instrument body. Therefore, when the reagent cartridge becomes
electrically charged, the electric charge is removed through the
aluminum foil.
[0125] Other instruments which were equipped mechanisms described
in table. 4, for example, the instruments without aluminum foil
were also prepared.
[Measurement of Luminescence]
[0126] The luminescent substrate solution of 70 .mu.L and the 70
.mu.L of H.sub.2O.sub.2 solution prepared as above were
automatically dispensed into the reactor tank of the reagent
cartridge in SphereLight 180, and then, luminescence of 140 .mu.L
in total volume of the reaction solution was measured. Measurement
was repeated 5 times (every 20 seconds in this Example) at the same
interval in the same reaction solution.
[Results]
[0127] Results are shown in Table 4 (Unit: cps). In Table 4, the
result obtained by measuring luminescence using the measuring
instrument in which countermeasure for preventing the influence of
the electric charge is not provided is determined as control. Also,
"versus Control" (%) is shown as a ratio of the average of signal
values obtained by measuring luminescence using measuring
instrument in which countermeasure for preventing the influence of
the electric charge is provided, to the average of the signal value
of control.
TABLE-US-00005 TABLE 4 Aluminum Foil at the without without with
undersufrace of photometry chamber Anti-static Tape on without with
without photometric chamber Number of Measurement 1 6,733 3,192
8,408 2 6,608 3,282 7,591 3 6,321 3,230 7,681 4 6,980 2,740 7,342 5
7,105 3,744 7,754 Average 6,749 3,238 7,755 versus Control 100% 48%
115%
[0128] As is clear from Table 4, background value can not be
suppressed even though the electric charge of the reagent cartridge
is removed by setting the aluminum foil at the undersurface of the
photometry chamber (The value of versus Control is high.). From
this result, it becomes clear that electrostatic charge of the
reagent cartridge does not affect on the increase of background
value. That is, it is understood from the result that background
value cannot be suppressed even though the grounding is set in the
photometry chamber. The other hand, background value can be
suppressed by attaching an anti-static tape in the photometry
chamber even without grounding.
Example 4
Effect of Exchanging an Atmosphere Surrounding a Reactor Tank in a
Photometry chamber with Gas Having a Constant Electric Charge
[0129] [Preparation of Reagents] The same reagents as in Example 1
were used.
[Measuring Instrument] Using SphereLight 180 which was automated
chemiluminescence enzyme immunoassay analyzers (Olympus Ltd.), an
air blowing tube was installed in its photometry chamber so that
atmospheric air was blown into the photometry chamber by a pump
(xx5510000, Millipore Co., Ltd).
[Measurement of Luminescence]
[0130] Measurement of luminescence was conducted by the same method
as in Example 1, except that countermeasure for preventing the
influence of the electric charge was performed by using the
above-described measuring instrument. That is, the external air was
continuously blown during measurement into the photometry chamber
by the pump, and the electric charge of the atmosphere surrounding
the reaction vessel in the photometry chamber was maintained
constant.
[0131] Measurement was repeated 4 times at the same interval (every
20 seconds in this Example) in the same reaction liquid.
[Results]
[0132] Results are shown in Table 5 (Unit: cps). In Table 5,
control means a result obtained by measuring luminescence under the
condition without blowing air (Countermeasure for preventing the
influence of the electric charge is not provided.).
[0133] Also, "versus Control" (%) is shown as a ratio of the
average of the signal values obtained by measuring luminescence
under air exchange, to the average of signal value of control.
TABLE-US-00006 TABLE 5 Number of Measurement Control Air Exchange
versus Control 1 4,940 3,317 2 5,184 3,556 3 5,189 3,506 4 5,230
3,295 Average 5,136 3,419 67%
[0134] As is clear from Table 5, the signal value (versus control
value) is 67% when the atmosphere surrounding the reaction vessel
in the photometry chamber is exchanged with the external air.
Therefore, it is understood that background value can be suppressed
by exchanging the atmosphere surrounding the reaction vessel in the
photometry chamber with the external air.
Example 5
Effect of Utilization of Neutralization Apparatus
[0135] [Preparation of Reagents] The same reagents as in Example 1
were used.
[Measuring Instrument] Using SphereLight 180 which was automated
chemiluminescence enzyme immunoassay analyzers (Olympus Ltd.), a
neutralization apparatus (SJ01 SJ020, Keyence Co., Ltd.) was
installed at the position so that electric charge surrounding the
reaction vessel of the reagent cartridge could be neutralized just
before measuring operation. And charge neutralization around the
reagent cartridge was carried out by using the apparatus.
[Measurement of Luminescence]
[0136] Luminescence was measured by the same method as Example 1,
except that the measuring instrument with charge neutralization
countermeasure was used.
[0137] Measurement was repeated 4 times (every 20 seconds in this
Example) at the same interval in the same reaction solution.
[Results]
[0138] Results are shown in Table 6 (Unit: cps). In Table 6,
"Control" means a result obtained by measuring luminescence by
using measuring instrument without a neutralization apparatus
(Countermeasure for preventing the influence of the electric charge
is not provided.). Also, "versus Control" (%) is shown as a ratio
of the average of signal values obtained by the measuring
luminescence using the measuring instrument with neutralization
apparatus is installed therein, to the average of the signal value
of control.
TABLE-US-00007 TABLE 6 Number of with Neutralization versus
Measurement Control Apparatus Control 1 6,084 2,942 2 6,946 3,687 3
6,948 3,279 4 6,885 3,837 Average 6,716 3,436 51%
[0139] As obvious from Table 6, the signal value is 51% when the
charge neutralization treatment is carried out for the reagent
cartridge using the neutralization apparatus. This means that
background value is suppressed by charge neutralization treatment
of the present invention.
Example 6
Measurement of an Analyte with the Prevention of the Influence of
the Electric Charge
[0140] [Preparation of Samples and Reagents]
[0141] As samples, 0, 0.01, 0.1, 2.5, 10 and 100 .mu.IU/ml of TSH
(Thyroid Stimulating Hormone) (Solvent: Diluting Solution of
Analyte, Wako Pure Chemical Industries, Ltd.) were prepared.
[0142] As reagents, SphereLight TSH III-measuring reagent kit (Wako
Pure Chemical Industries, Ltd.) with the following constitution was
prepared.
[0143] Solid Phase: Polystyrene beads (1/8 inches) immobilized with
an anti-TSH monoclonal antibody,
[0144] A Labeled Antibody Solution: A peroxidase (POD)-labeled
anti-TSH monoclonal antibody,
[0145] A Diluting Solution of Analyte: A diluting solution of
analyte exclusive for SphereLight 180,
[0146] A Luminescent Substrate Solution: A solution of 5 mM luminol
(a buffer solution exclusive for SphereLight 180, pH 8.5, Wako Pure
Chemical Industries, Ltd.),
[0147] A H.sub.2O.sub.2 Solution: An aqueous solution of 0.02%
H.sub.2O.sub.2, (an acid buffer solution exclusive for SphereLight
180, pH 3.0, Wako Pure Chemical Industries, Ltd.).
[Measuring Instrument]
[0148] Using SphereLight 180 which was automated
chemiluminescence's enzyme immunoassay analyzers (Olympus Ltd.),
following instrument was used under condition as described
below.
[0149] (1) The instrument in which countermeasure for preventing
the influence of the electric was not provided,
[0150] (2) The instrument which the anti-static tapes attached
transversely on four inside wall surfaces of the photometry
chamber, in the same way as Example 1 (A preventing method a),
[0151] (3) The instrument which the anti-static tapes attached
transversely on four inside wall surfaces of the photometry
chamber, and moreover, a neutralization apparatus (SJ01 SJ020,
Keyence Co., Ltd.) installed at the position so that electric
charge surrounding the reaction vessel of the reagent cartridge
could be neutralized, in order to electric charge surrounding the
reagent cartridge was neutralized (A preventing method b).
[Measurement of Luminescence]
[0152] The TSH sample of 50 .mu.L with each concentration prepared
by the above method and 90 .mu.L of the diluting solution of
analyte were added into the reaction vessel holding the above beads
immobilized with the anti-TSH antibody, mixed, and reacted at
37.degree. C. for 7 minutes. After the reaction, the beads were
washed, and 140 .mu.L of the POD-labeled anti-TSH antibody solution
was added thereto, and mixed, followed by allowing a reaction to
take place at 37.degree. C. for 7 minutes. After the reaction, the
beads were washed, and the luminescent substrate solution of 70
.mu.l, and 70 .mu.L of the H.sub.2O.sub.2 solution were added
thereto and mixed. After mixing, luminescence in the reaction
solution was measured.
[Results]
[0153] Results are shown in Table 7 (Unit: cps)
[0154] In Table 7, "S/N" means the radio of "a value obtained by
measuring the luminescence of the sample containing each
concentration of TSH" to "a value obtained by measuring the
luminescence of the sample which does not contain TSH by the same
method as described above.
TABLE-US-00008 TABLE 7 Sample, without Preventing Method Preventing
Method a Preventing Method b (.mu.IU/mL) Data Average S/N Data
Average S/N Data Average S/N 0 17,745 6,434 5,479 17,991 17,868
1.00 6,413 6,424 1.00 5,580 5,530 1.00 0.01 19,589 9,185 8,535
19,607 19,598 1.10 9,051 9,118 1.42 8,563 8,549 1.55 0.1 54,249
44,794 43,022 51,188 52,719 2.95 43,292 44,043 6.86 43,822 43,422
7.85 2.5 1,341,577 1,404,537 1,384,086 1,398,990 1,370,284 76.69
1,314,811 1,359,674 211.67 1,320.636 1,352,361 244.57 10 5,569,448
5,387,652 5,594,896 5,286,842 5,428,145 303.79 5,235,523 5,311,588
826.90 5,291,060 5,442,978 984.35 100 31,232,604 29,786,674
29,248,316 33,460,552 32,346,578 1,810.31 31,718,816 30,752,745
4,787.54 31,863,476 30,555,896 5,525.98
[0155] As is clear from Table 7, sensitivity improvement
(improvement of S/N ratio) is recognized in luminescence
measurement by carrying out the method for preventing the influence
of the electric charge of the present invention. Also, when two
methods for preventing are combined (a preventing method b), better
effect of sensitivity improvement (improvement of S/N ratio) is
obtained. Therefore, it is suggested that the effect of sensitivity
improvement is further enhanced with combination of appropriate
method for preventing the influence of the electric charge in the
present invention.
Example 7
Suppression of Signal Value Variation Caused by Environmental
Change
[0156] Using the same reagents as in Example 1, measurements were
carried out for different three days (Environment A, Environment B
and Environment C). Measurement was repeated 4 times (every 20
seconds in this Example) at the same interval in the same reaction
solution.
[0157] Measurements were carried out by using the instrument
attached the anti-static tapes transversely on four inside wall
surfaces of the photometry chamber, similarly as in Example 1. The
measurement using the instrument without attachment of the
anti-static tapes (Countermeasure for preventing the influence of
the electric charge was not provided.) was carried out, and the
obtained result was determined as control.
[0158] Results are shown in Table 8-1 and Table 8-2, and the former
shows a result of control, and the latter shows result obtained by
the measurement using the instrument which the anti-static tape
attached thereon, respectively. In Table 8-1 and Table 8-2, the
average of four times measured values obtained at the first
measurement day (Environment A) was determined as 100, and
"/Environment A" means a relative percentage (%) of measured value
in each environment (A to C) to the determined values which
described above.
TABLE-US-00009 TABLE 8-1 Number of Measurement Environment A
Environment B Environment C 1 4,069 15,155 20,745 2 3,943 14,379
20,991 3 3,935 14,681 21,450 4 4,291 14,934 22,948 Average 4,060
14,787 21,534 /Environment A 100% 364% 530%
TABLE-US-00010 TABLE 8-2 Number of Measurement Environment A
Environment B Environment C 1 4,492 5,808 6,749 2 3,492 5,014 3,479
3 3,700 5,501 5,843 4 3,820 5,258 5,580 Average 3,876 5,395 6,163
/Environment A 100% 139% 159%
[0159] As is clear from the result shown in Table 8-1 and Table
8-2, when the measurement is carried out by using the instrument in
which the countermeasure for preventing the influence of the
electric charge is not provided, obtained measurement value is
varied significantly depending the measurement date (Environmental
variation) (Table 8-1). Contrary to that, when the measurement is
carried out by using the instrument which the anti-static tape is
attached thereon, the obtained measurement value are not varied so
much even though the measurement day is different, in other word,
even in environmental variation (Table 8-1), and this means that
stable measurement is achieved by the present invention.
Example 8
Improvement of Reproducibility of Measured Value by Charge Removal
Countermeasure
[0160] Using the same measuring kit as in Example 6, luminescence
measurement in samples including 0.02 and 0.1 .mu.IU/mL of TSH was
carried out. Measurement was repeated 10 times (every 20 seconds in
this Example) at the same interval in the same reaction
solution.
[0161] The measurement was carried out by using an instrument
attached the anti-static tapes transversely on four inside wall
surfaces of a photometry chamber (with charge removal
countermeasure), similarly as in Example 1, and an instrument
without attachment of the anti-static tape (without charge removal
countermeasure).
[0162] Results are shown in Table 9.
TABLE-US-00011 TABLE 9 without Charge Removal with Charge Removal
Countermeasure Countermeasure TSH TSH TSH TSH 0.02 .mu.IU/mL 0.1
.mu.IU/mL 0.02 .mu.IU/mL 0.1 .mu.IU/mL 1 24,455 53,054 13,961
42,452 2 20,696 52,343 13,336 43,237 3 21,923 50,918 13,425 41,334
4 22,856 48,184 13,865 42,534 5 22,235 55,788 12,672 42,755 6
18,807 49,932 13,209 40,838 7 20,656 52,404 13,084 40,881 8 20,415
50,944 12,949 41,640 9 18,677 48,801 13,666 40,581 10 17,709 51,214
13,868 42,243 Average Value 20,843 51,358 13,404 42,880 SD Value
2,084 2,201 434 946 CV Value 10.0% 4.3% 3.2% 2.3%
[0163] As is clear from Table 9, the measured value obtained by
using the instrument with charge removal countermeasure shows less
variation of measured value, and therefore, reproducibility of
measured value is improved.
Example 9
Investigation on a Method for Shutting an Opening Part of a
Reaction Vessel Holding a Solution
[0164] [Preparation of Reagents] The same reagents as in Example 1
were used.
[0165] [Measuring Instrument]
[0166] SphereLight 180 which was automated chemiluminescence enzyme
immunoassay analyzer (Olympus Ltd.) was used similarly as in
Example 1. To the instrument, countermeasure for preventing the
influence of the electric charge was not provided.
[Measurement of Luminescence]
[0167] Measurement of luminescence was carried out under the
following condition. Measurement was repeated 5 times (every 20
seconds in this Example) at the same interval in the same reaction
solution. Incidentally, the reagent cartridge used for the
measurement has a concave portion to dispense the reagent solution,
and it is called as a reactor tank (or called as a measurement
cell).
(1) The above-prepared reagents were dispensed and mixed into the
reactor tank of the reagent cartridge (Total volume of reaction
solution was 140 .mu.L), and then, it was set in SphereLight 180
and luminescence was measured. (2) After measurement of (1), the
reagent cartridge was collected, and the opening part of the
reactor tank was sealed with a cellophane tape (shut). And then, it
was set in SphereLight 180 again, and luminescence was measured.
(3) After measurement of (2), the reagent cartridge was collected,
and the cellophane tape was peeled off, it was set in SphereLight
180 again, and luminescence was measured. (4) After measurement of
(3), the reagent cartridge was collected, and the opening part of
the reactor tank of the reagent cartridge was sealed with a
cellophane tape (shut). And then, it was set in SphereLight 180
again, and luminescence was measured.
[0168] [Results]
[0169] Results are shown in Table 10 (Unit: cps). In Table 10, the
result obtained by measuring luminescence under condition without
sealing the reactor tank is determined as control. And "versus
Control" (%) is shown as a ratio of the average of signal values
obtained by each measurement method, to the average of the signal
value of control.
TABLE-US-00012 TABLE 10 Measuring Method (1) (2) (3) (4) Sealing of
Reaction -- Sealed -- Sealed Tank Number of Measurement 1 1909 466
1981 524 2 2008 449 2285 469 3 1983 462 2113 513 4 2351 415 2135
513 5 2280 407 2211 523 Average 2106 440 2145 508 versus Control
100% 21% 102% 24%
[0170] As is clear from Table 10, background value is suppressed by
sealing the opening part of the reactor tank with the cellophane
tape whereby blocking the contact of the atmosphere in the
photometry chamber with the solution in the reaction vessel
(2).
[0171] After background value is confirmed to be suppressed, the
cellophane tape is peeled off and again measurement is carried out,
and the background value is increased again (3). However when
luminescence is measured after sealing the opening part of the
reactor tank again with the cellophane tape, the background value
is suppressed again (4). From these results, it is suggested that
background value varies by contacting atmosphere in the photometry
chamber with the reagent.
Example 10
Investigation on Sheet to Shut the Opening Part of a Reaction
Vessel
[0172] [Preparation of Reagents] The same reagents as in Example 1
were used.
[Measuring Instrument]
[0173] SphereLight 180 which was automated chemiluminescence enzyme
immunoassay analyzers (Olympus Ltd.) was used. To the instrument,
countermeasure for preventing the influence of the electric charge
was not provided.
[0174] [Measurement of Luminescence]
[0175] The reagents prepared above were dispensed and mixed into
the reactor tank of the reagent cartridge (total volume of reaction
solution was 140 .mu.L) and the opening part of the reactor tank
was sealed (shut) with an aluminum seal or a cellophane tape, and
then, it was set in SphereLight 180, and luminescence was measured.
Measurement without sealing the opening part of the reactor tank
was carried out, and obtained result determined as control.
Measurement was repeated 5 times (every 20 seconds in this Example)
at the same interval in the same reaction solution.
[Results]
[0176] Results are shown in Table 11 (Unit: cps). In Table 11,
"versus Control" (%) is shown as a ratio of the average of the
signal values obtained by measuring luminescence using the reagent
cartridge with the opening part of the reactor tank sealed, to the
average of signal values of control.
TABLE-US-00013 TABLE 11 Seal of Reactor Tank without covered
covered Seal Type without Aluminum Seal Cellophane Tape Number of
Measurement 1 1,697 604 459 2 1,167 437 451 3 1,425 618 408 4 1,315
572 449 5 1,039 589 489 Average 1,329 564 451 versus Control 100%
42% 34%
[0177] As is clear from Table 11, background value can be
suppressed by sealing (shutting) the opening part of the reactor
tank with blocking the contact of the atmosphere surrounding the
photometry chamber with the solution in the reaction vessel.
[0178] Also, background value can be suppressed at nearly the same
level irrespective of whether shutting is carried out with the
aluminum seal or the scotch seal.
Example 11
Investigation on a Method for Covering the Surface of a Reaction
Solution with a Water-Insoluble Liquid
[0179] [Preparation of Reagents] The same reagents as in Example 1
were used.
[Measurement of Luminescence]
[0180] The above-prepared reagents were dispensed and mixed into
the reactor tank of the reagent cartridge (Total volume of reaction
solution was 140 .mu.L), and then silicon oil (Shin-Etsu Silicon
Co., Ltd) was funneled gently along the inside wall surface of the
reactor tank using a pipette to the surface of the mixed reaction
solution. And the surface of the reaction solution was covered with
silicon oil to provide oil film.
[0181] And then, the reagent cartridge was set in SphereLight 180
and luminescence was measured. Similar measurement without covering
the surface of the reaction solution was carried out, the obtained
result was determined as control. Measurement was repeated 5 times
(every 20 seconds in this Example) at the same interval in the same
reaction solution.
[Measuring Instrument]
[0182] SphereLight 180 which was automated chemiluminescence enzyme
immunoassay analyzer (Olympus Ltd.) was used. One instrument
attached the anti-static tapes (25 mm wide, Japan Vilene Co., Ltd.)
transversely on four inside wall surfaces of a photometry chamber,
and the other instrument without attachment of the anti-static tape
was prepared.
[Results]
[0183] Results are shown in Table 12 (Unit: cps). In Table 12,
"versus Control" (%) is shown as a ratio of the average of the
signal values obtained by measuring luminescence in the solution
which treated its surface of reaction solution with silicone oil
film, to average of signal values of control.
TABLE-US-00014 TABLE 12 Anti-static Tape on without without with
with Photometry Chamber Silicon Oil Film without with without with
Number of 1 1,765 802 1,615 673 Measurement 2 2,691 855 1,791 751 3
2,097 864 1,467 783 4 2,883 987 1,600 929 5 3,733 1,037 2,175 824
Average 2,634 909 1,730 792 versus Control 100% 35% 66% 30%
[0184] As is clear from Table 12, background value can be
suppressed by covering the surface of the reactor tank with silicon
oil and by blocking the contact of an atmosphere surrounding the
photometry chamber with the solution in the reaction vessel.
BRIEF DESCRIPTION OF THE DRAWINGS
[0185] [FIG. 1] A view showing a frame format of the photometry
chamber of SphereLight 180 which is automated chemiluminescence
enzyme immunoassay analyzer used in pattern 5 of Example 2. In FIG.
1, four inside wall surfaces of the photometry chamber are shown as
A, B, C and D, respectively. Also, the attached position of the
anti-static tape, the position of the photometry chamber, the
position of the photoelectron multiplier against the photometry
chamber, and setting position of the reagent cartridge holding
reaction solution are shown.
[0186] [FIG. 2] A view showing the frame format of the photometry
chamber of SphereLight 180 which is automated chemiluminescence
enzyme immunoassay analyzer used in Example 3. In FIG. 2, four
inside wall surfaces of the photometry chamber are shown as A, B, C
and D, respectively. Also, the set position of aluminum foil in the
photometry chamber is shown by oblique lines.
* * * * *