Method for effecting heterogeneous immunological analysis

Method for generating an automatic chemical analyzer for measuring given substances in samples in accordance with an enzyme-immunoassay including a turntable rotated intermittently at a given pitch and holding a number of reaction tubes arranged equidistantly along a periphery of the turntable to define a circular reaction line; a carrier supply device for supplying carriers into reaction vessels one by one at a given position in the reaction line, the carrier having given antibody or antigen fixed thereto; a sample delivery device for pouring given amounts of samples into reaction vessels at a given position in the reaction line; a washing device for washing reaction vessels and carriers contained therein to effect B-F separation; a color reagent delivery device for pouring given amounts of a color reagent into reaction vessels to form test liquids; a colorimeter for photometering the test liquids; and a carrier discharge device for removing carriers out of reaction vessels. For respective samples, a reaction vessel is passed through the washing device by a plurality of times to perform a plurality of washing operations including the B-F separation.

BACKGROUND OF THE INVENTION 
The present invention relates generally to immunological analysis and more 
particularly to a method of analyzing automatically given substances in 
samples due to an immunological reaction of antigen and antibody. 
At present, due to progress in medical treatment, very small amounts of 
biological substances in samples can be analyzed and this contributes to 
early diagnosis for various diseases. For instance, malignant tumors such 
as .alpha.-fetoprotein and carcinoembryonic antigen, diseases in abnormal 
secretory of hormone such as insulin and thyroxine, and immunological 
diseases such as immunoglobulin can be diagnosed in early stages and 
further the monitoring after treatments for these diseases can be carried 
out reliably. Moreover, the measurement of incomplete antigens, i.e. low 
molecular hapten of medical substances contributes to development of a 
plan of medication. 
Many biological substances are analyzed in an immunological manner by 
utilizing the antigen-antibody reaction and various methods for effecting 
the immunological analysis have been developed. For instance, existence or 
non-existence of agglinated clots of antigen-antibody compound formed by 
the antigen-antibody reaction is detected by agglutination method, 
sedimentry method, nephelometry method, etc. to analyze desired biological 
substances. However, in the known methods, since the sensitivity is low, a 
large amount of antigen-antibody compound is required and only qualitative 
analyses or quasi-quantitative analyses can be performed. In order to 
avoid such a drawback, there have been further proposed the following 
methods. In one of the known methods, antigen or antibody is bound with 
carbon or synthetic resin fine particles which are then subjected to the 
antigen-antibody reaction with the biological substances to be analyzed 
and the substances are detected by means of the agglutination method or 
nephelometry method. In another known method, antigen-antibody compounds 
are detected at a high sensitivity by using antigen or antibody marked 
with labeling material such as radioisotope, fluorescent material, 
luminescent material and enzyme. However, since the former method is 
inferior to the latter method in the sensitivity, recently the latter 
method using the high sensitivity labeling substance has been 
predominantly adopted. 
The analytic methods using the markers are classified into 
radio-immuno-assay using radioisotope tracers, fluorescent-immuno-assay 
using fluorescent labeling material, and enzyme-immuno-assay using enzyme 
markers. Among these methods, the enzyme-immuno-assay has been 
particularly developed owing to the reason that it does not require 
special installation and measuring technique and can be performed easily 
by using commonly developed colorimeters. The enzyme-immuno-assay is 
further classified into homogeneous enzyme-immuno-assay and heterogeneous 
enzyme-immuno-assay. In the homogeneous analysis, a variation in activity 
of labeling enzyme due to existence or non-existence of the immunological 
reaction is directly measured to detect substances to be analyzed. In the 
heterogeneous analysis, use is made of insoluble carriers such as glass 
beads or synthetic resin particles on which antigen or antibody has been 
fixed, enzyme-labeled antigen or antibody bound with the antibody or 
antigen fixed on the carriers and free enzyme-labeled antigen or antibody 
not bound with the antibody or antigen on the carriers are separated from 
each other by washing treatment, and then an activity of labeling enzyme 
is detected to measure a quantity of substances to be analyzed. 
Hereinbelow, the process for separating the bound antigen or antibody and 
the free antigen or antibody from each other is termed as B-F or 
bound-free separation for the sake of simplicity. Although the homogeneous 
analysis can be performed by simple processes, it can analyze only the low 
molecular hapten such as medical substances, but cannot analyze high 
molecular biological substances. Contrary to this, in the heterogeneous 
analysis, although the washing process is required for effecting the B-F 
separation, it can be applied to any kinds of low and high molecular 
substances. Therefore, recently the heterogeneous enzyme-immuno-assay has 
been generally adopted. 
In the heterogeneous enzyme-immuno-assay, there have been developed 
competitive method and sandwich method. Now these methods will be 
explained with reference to the drawings. 
FIG. 1 illustrates successive steps of the competitive method. Given 
antigen or antibody which reacts with antibody or antigen substances 2 of 
a sample has been previously fixed to an outer surface of a insoluble 
carrier 1. At first, the antigen-antibody reaction is carried out between 
the antigen or antibody fixed onto the carrier 1 and the antibody or 
antigen 2 in the sample as well as a labeled reagent 3 which has been 
prepared by labeling substances same as the substances 2 to be analyzed 
with enzyme marker. Then, a washing process is carried out to effect the 
B-F separation between the substance 2 and labeled reagent 3 bound with 
the carrier 1 due to the antigen-antibody reaction and free substances 2 
and reagent 3 which are not bound with the carrier 1. Next, a color 
reagent which selectively reacts with the labeling enzyme is added and a 
reaction liquid is colorimetered to detect the enzyme activity of the 
labeling enzyme. 
FIG. 2 shows successive steps of the sandwich method in which use is made 
of an insoluble carrier 5 having fixed thereto antibody or antigen which 
is reactive with antigen or antibody substances in a sample to be tested. 
At first, the carrier 5 and the sample 6 are mixed to effect the 
antigen-antibody reaction between the substances 6 in the sample and the 
antibody or antigen fixed to the carrier 5. Then, the B-F separation is 
carried out by means of the washing step. Next, a labeled reagent 7 is 
added to effect the antigen-antibody reaction. The labeled reagent is 
prepared by marking with enzyme substance selectively reacting with the 
substance 6 to be analyzed. Then, after the B-F separation is effected 
again, a color reagent reacting with the labeling enzyme in the labeled 
reagent 7 is added and a test liquid thus obtained is colorimetered to 
detect the activity of the labeling enzyme. 
As explained above, in the heterogeneous immuno-assay the B-F separation 
has to be carried out once in the competitive method and twice in the 
sandwich method during the analysis for respective sample and further if a 
reaction vessel for effecting the antigen-antibody reaction is used 
repeatedly, there must be further provided a step for washing the reaction 
vessel after the end of analysis for a sample, but before the start of 
analysis for another sample. In case of automating the enzyme-immuno-assay 
requiring at least two washing steps including the B-F separation, 
separate washing devices may be provided at different positions. However, 
then an automatic analyzer is liable to be large in size, complex in 
construction and expensive in cost. This disadvantage will also appear in 
automatic analyzer effecting radio-immuno-assay and 
fluorescent-immuno-assay. 
In a Japanese Patent Application Laid-open Publication No. 74,662/82 
published on May 10, 1982, there is disclosed an automatic enzyme-immuno 
assay apparatus in which the B-F separation is effected by removing a 
carrier having antigen-antibody compound bound thereto from a reaction 
vessel to another reaction vessel. It is apparent such an analyzer becomes 
large in size and requires a special mechanism for transporting the 
carrier. Further, the efficiency of analysis is low and a number of 
samples could not be processed promptly. In a Japanese Patent Application 
Laid-open Publication No. 124,254/82 published on Aug. 3, 1982, there is 
described an automatic analyzer for effecting an immunological analysis in 
which the B-F separation is performed by rotating a rotor on which 
reaction vessels having carriers contained therein are arranged. In this 
analyzer, since the excess liquid is discharged in all directions due to 
the centrifugal force, the treatment of the discharged liquid becomes 
cumbersome. Further, since the reaction vessels containing the carriers 
are repeatedly used, a special treatment for releasing the antigen or 
antibody from the carriers must be effected between successive analyses. 
SUMMARY OF THE INVENTION 
The present invention has for its object to provide a method for effecting 
automatically an immunological analysis which can be carried out stably 
and effectively by a small, simple and inexpensive apparatus. 
According to the invention, a method of automatically analyzing given 
substances in samples in an immunological manner comprises: 
transporting a number of reaction vessels containing carriers onto which 
given antibody or antigen has been fixed or a number of reaction vessels 
having given antibody or antigen fixed onto at least a part of inner 
walls, along a reaction line; 
delivering samples and labeled reagents into the reaction vessels to 
initiate antigen-antibody reaction; 
effecting a B-F separation by separating antigen or antibody bound with the 
carriers or reaction vessels and free antigen or antibody from each other 
by means of washing; 
measuring the given substances in the samples with the aid of labeling 
substances of the labeled reagent; and 
discharging the carriers or the reaction vessels out of the reaction line. 
The present invention also relates to an automatic analyzer for carrying 
out the above method and has for its object to provide a novel and useful 
automatic analyzer which can be made simple in construction, small in size 
and inexpensive in cost. 
According to the invention, an automatic analyzer for analyzing given 
substances in samples in an immunological manner comprises: 
means for transporting a number of reaction vessels along a given reaction 
line; 
means for supplying carriers into reaction vessels at a given position in 
the reaction line, said carriers having given antibody or antigen fixed 
thereto; 
means for supplying given amounts of samples into the reaction vessels at a 
given position of the reaction line; 
means for delivering given amounts of a labeled reagent into the reaction 
vessels at given positions in the reaction line; 
means for washing the reaction vessels and carriers to effect a B-F 
separation for separating antigen or antibody bound with the carriers and 
free antigen or antibody; 
means for measuring the given substances in the samples with the aid of 
labeling substances of the labeled reagent; and 
means for discharging the carrier from the reaction line. 
The present invention further relates to a reaction vessel for use in the 
above method and has for its object to provide a novel reaction vessel for 
use in the immunological analysis. 
According to the invention, a reaction vessel for use in a method of 
analyzing given substances in samples with the aid of given antibody or 
antigen fixed to solid substance and a labeled reagent having given 
antibody or antigen labeled with given substance, by supplying reaction 
vessels into a reaction line, delivering the samples and reagent into the 
reaction vessels, washing the reaction vessels to effect a B-F separation, 
measuring the given substances in the samples with the aid of the labeled 
reagent and discharging the reaction vessels from the reaction line, 
comprises: 
a main body having an opening at its top; and 
a given antibody or antigen layer fixed onto at least a part of an inner 
wall of the main body.

DESCRIPTION OF THE PREFERRED EMBODIMENTS 
FIG. 3 is a schematic view showing an embodiment of the enzyme-immuno-assay 
automatic analyzer according to the invention which performs the sandwich 
method explained above with reference to FIG. 2. In the present 
embodiment, there is provided a single reaction line to effect an analysis 
for a single test item. As a reaction vessel use is made of a U-shaped 
tube 11 having large and small mouth portions 11a and 11b. On a turntable 
12 are arranged equidistantly twenty four U-shaped tubes 11 along a 
periphery of the turntable. The turntable 12 is intermittently rotated in 
a direction shown by an arrow a at a given period of, for example 15 
seconds, while the U-shaped tubes 11 are dipped into a thermostat 10 (see 
FIG. 4). Positions at which the U-shaped tubes 11 are stopped due to the 
stepwise rotation of the turntable 12 are denoted as S.sub.1 to S.sub.24. 
Hereinafter, the term "pitch" denotes the angular distance increment 
between adjacent ones of the equidistantly spaced vessels 11 on the 
turntable 12. In the present embodiment, into a U-shaped tube 11 
positioned at S.sub.1 is delivered a sample from a sample cup 15 which is 
situated just at a sample sucking position of a sampler 14 by means of a 
sample delivery device 13. The sampler 14 holds twenty four sample cups 15 
arranged equidistantly along a disc which is rotated in a direction b in 
synchronism with the rotation of the turntable 12. In a U-shaped tube 11 
in S.sub.3 is selectively delivered an enzyme reagent 17 corresponding to 
substances in samples to be tested, by means of a reagent delivery device 
16. In a U-shaped tube 11 situated at S.sub.4 is poured a color reagent 19 
with the aid of a reagent delivery device 18. Into a U-shaped reaction 
tube 11 is supplied a carrier 21 such as a synthetic resin particle or 
glass bead from a carrier supply device 21. It should be noted that the 
carrier 21 has a diameter smaller than an inner diameter of the large 
mouth portion 11a of the U-shaped tube 11, but is larger than an inner 
diameter of the small mouth portion 11b. On an outer surface of the 
carrier 21 there has been previously fixed antibody or antigen which 
causes the antigen-antibody reaction with antigen or antibody substance in 
the sample to be tested. Further, in the carrier supply device 20, the 
carriers 21 are wetted with a buffer solution. A reaction liquid in a 
U-shaped tube 11 at a position S.sub.19 is sucked into a colorimeter 22, 
and a carrier 21 contained in a U-shaped tube 11 at a position S.sub.20 is 
removed therefrom by means of a carrier discharge device 23. Into a 
U-shaped tube 11 at a position S.sub.22 is supplied a washing liquid such 
as ion exchange water, buffer solution for immunological analysis, 
physiological saline solution, etc. In a U-shaped tube situated at a 
position S.sub.24 is selectively delivered a buffer solution 26 by means 
of a buffer solution delivery device 25. At positions S.sub.2 to S.sub.5, 
a stirring air pump 27 can be detachably connected to small mouth portions 
11b of U-shaped tubes 11, and at positions S.sub.22 and S.sub.23, a 
discharge pump 28 can be detachably connected to small mouth portions 11b 
of U-shaped tubes 11. 
Now, the operation of the automatic analyzer shown in FIG. 3 will be 
explained also with reference to FIGS. 4A to 4D. 
During a first revolution of the turntable 12, at the position S.sub.17, a 
carrier 21 wetted with the buffer solution is supplied in a U-shaped tube 
11 via its large mouth portion 11a as shown in FIG. 4A. Then, at the 
position S.sub.22, the washing liquid is intermittently poured into the 
U-shaped tube 11 from the large mouth portion 11a just like a shower by 
means of the washing pump 24 and at the same time, the washing liquid is 
sucked out of the tube 11 via the small mouth portion 11b by means of the 
discharge pump 28. Next, at the position S.sub.23, any washing liquid 
remaining in the tube 11 is discharged by the discharge pump 28. In this 
manner, the U-shaped reaction tube 11 is washed and at the same time, the 
buffer solution on the carrier 21 is removed. This ensures that an amount 
of the buffer solution 26 to be supplied by the buffer solution delivery 
device 25 can be made to be a given constant value. 
Then, as illustrated in FIG. 4B, at the position S.sub.24 a given amount of 
the buffer solution 26 is delivered into the U-shaped tube 11 via its 
large mouth portion 11a by means of the delivery device 25. Then, at the 
position S.sub.1 a given amount of a sample is delivered by means of the 
sample delivery device 13 into the tube 11 via its large mouth portion 11a 
from a sample cup 15 situated at the sample sucking position of the 
sampler 14. Next, at the positions S.sub.2, S.sub.3, S.sub.4 and S.sub.5, 
air streams are supplied into the U-shaped tube 11 from its small mouth 
portion 11b by means of the air pump 27 to stir the buffer solution and 
sample in the tube 11. In this manner, a first antigen-antibody reaction 
is caused to proceed. It should be noted that the carrier supply device 13 
and sampler 14 are made inoperative after being once operated for 
respective U-shaped tubes. 
During a second revolution of the turntable 12, at the position S.sub.22, 
the liquid in the tube 11 is sucked via the small mouth portion 11b by the 
discharge pump 28 and at the same time, the washing liquid is 
intermittently poured into the tube via its large mouth portion 11a by 
means of the washing pump 24. The washing liquid remaining in the tube is 
discharged at the positions S.sub.22 and S.sub.23 as shown in FIG. 4B. In 
this manner, the U-shaped tube 11 and the carrier 21 contained therein are 
fully washed to effect a first B-F separation. Then, at the position 
S.sub.3, a given amount of the enzyme-labeled reagent 17 is delivered into 
the U-shaped tube 11 via its large mouth portion 11a by the reagent 
delivery device 16 as illustrated in FIG. 4C. The reagent and carrier are 
stirred sufficently at the positions S.sub.3, S.sub.4 and S.sub.5 by 
supplying the air streams from the small mouth portion 11b with the aid of 
the air pump 27 to effect a second antigen-antibody reaction. 
During a third revolution of the turntable 12, at the positions S.sub.22 
and S.sub.23, the U-shaped tube 11 and carrier are washed by means of the 
washing pump 24 and discharge pump 28 to perform a second B-F separation. 
Next, as shown in FIG. 4D, a given amount of the color reagent 19, i.e. 
the enzyme substrate reagent, is delivered into the U-shaped tube 11 by 
the reagent delivery device 18. Then, the color reagent and carrier are 
stirred by means of the air pump 27 to produce a reaction of the color 
reagent 19 with the labeling enzyme of the enzyme-labeled reagent 17 bound 
with the carrier 21. 
In a fourth revolution of the turntable 12, at the position S.sub.19 a 
reaction liquid in the U-shaped tube 11 is sucked into the colorimeter 22 
to effect the colorimetric measurement. As depicted in FIG. 4D, the 
colorimeter 22 comprises a flow cell 22a through which the reaction liquid 
is flown, and light source 22b and detector 22c arranged on respective 
sides of the flow cell 22a. Light emitted from the light source 22b is 
projected into the flow cell 22a via an interference filter 22d and light 
transmitted through the flow cell 22a is received by the detector 22c by 
means of a light guide 22e. 
At the position S.sub.20, the carrier 21 is sucked out of the U-shaped tube 
11 via its large mouth portion 11a by the carrier discharge device 23. At 
the position S.sub.22, the washing liquid is supplied into the U-shaped 
tube 11 via its large mouth portion 11a like a shower and the wash liquid 
is sucked out of the tube via its small mouth portion 11b. The wash liquid 
remaining in the tube is discharged at the position S.sub.23. In this 
manner, the U-shaped tube 11 is prepared for a next supply of a carrier. 
As explained above in detail, in the present embodiment since the U-shaped 
reaction tube 11 is repeatedly passed through the washing device 
comprising the washing pump 24 and discharge pump 28 to effect the washing 
including the B-F separation repeatedly, the whole analyzer can be made 
small in size and simple in construction. 
FIG. 5 is a schematic view showing another embodiment of the automatic 
analyzer for effecting the enzyme-immuno-assay according to the invention. 
Portions similar to those shown in FIG. 3 are denoted by the same 
reference numerals used in FIG. 3. In the present embodiment, prior to the 
supply of a carrier 21 into a U-shaped tube 11, a given amount of a buffer 
solution 32 has been delivered into the tube at a position S.sub.16 by 
means of a second buffer solution delivery device 31. The remaining 
construction and operation are entirely the same as those of the previous 
embodiment shown in FIG. 3. When the buffer solution 32 has been delivered 
into the tube 11, the carrier 21 can be supplied into the tube at the 
position S.sub.17 with a minimum shock. It should be noted that it is not 
necessary to control precisely the delivery amount of the buffer solution 
32, because the U-shaped tube 11 and carrier 21 are washed at the position 
S.sub.22. 
FIG. 6 is a schematic view illustrating an embodiment of the automatic 
analyzer according to the invention, in which the enzyme-immuno-assay is 
performed by the competitive method. Also in the present embodiment 
portions similar to those shown in FIG. 3 are denoted by the same 
reference numerals used in FIG. 3. In this embodiment, the delivery of the 
color reagent at the position S.sub.4 and the mixing at the position 
S.sub.5 are removed. At the position S.sub.3, a given amount of a color 
reagent 36 instead of the enzyme-labeled reagent is delivered by means of 
a reagent delivery device 35 and at the position S.sub.24, a given amount 
of an enzyme-labeled reagent 38 instead of the buffer solution is 
delivered by an enzyme-labeled reagent delivery device 37, said 
enzyme-labeled reagent 38 is prepared by marking with enzyme the same 
substance as that in a sample to be analyzed. The remaining construction 
of the analyzer in the present embodiment is entirely the same as that of 
the embodiment illustrated in FIG. 3. 
Now the operation of the enzyme-immuno-assay automatic analyzer illustrated 
in FIG. 6 will be explained in detail also with reference to FIGS. 7A to 
7C. 
During the first revolution of the turntable 12, at the position S.sub.17, 
a carrier 21 wetted with the buffer solution is supplied from the carrier 
supply device 20 into a U-shaped tube 11 as shown in FIG. 7A. Then, at the 
position S.sub.22, the washing liquid is poured intermittently into the 
tube 11 like a shower by the washing pump 24 and the washing liquid 
remaining in the tube 11 is sucked out of the tube at the position 
S.sub.23 by means of the discharge pump 28 via the small mouth portion 11b 
of the U-shaped tube. Next, as illustrated in FIG. 7B, at the position 
S.sub.24 a given amount of the enzyme-labeled reagent 38 is delivered into 
the U-shaped tube 11 from its large mouth portion 11a by means of the 
reagent delivery device 37 and then at the position S.sub.1, a given 
amount of a sample in a sample cup 15 in the sampler 14 is delivered into 
the U-shaped tube 11. Next, at the positions S.sub.2 to S.sub.4 the air 
streams are flown through the U-shaped tube 11 from its small mouth 
portion 11b to its large mouth portion 11a with the aid of the air pump 27 
to mix the carrier 21, enzyme-labeled reagent 38 and sample with one 
another to produce the antigen-antibody reaction. It should be noted the 
carrier supply device 20, reagent delivery device 37, sample delivery 
device 13 and sampler 14 are kept inoperative once being operated for 
respective U-shaped tubes. 
During a second revolution of the turntable 12, at the position S.sub.22 
the reaction liquid in the U-shaped tube 11 is sucked out of the tube by 
the discharge pump 28 and at the same time, the washing liquid is poured 
into the tube 11 and the washing liquid remaining in the tube 11 is 
discharged at the positions S.sub.22 and S.sub.23 by the pump 28 to effect 
the B-F separation. 
Then, as shown in FIG. 7C, at the position S.sub.3 a given amount of the 
color reagent 36 is delivered by the reagent delivery device 35 into the 
U-shaped tube 11. Next, at the positions S.sub.3 and S.sub.4 the air 
streams are passed through the tube with the aid of the air pump 27 to 
stir the carrier 21 and color reagent 36 to produce the reaction. 
In a third revolution of the turntable 12, at the position S.sub.19 the 
reacted liquid in the U-shaped tube 11 is sucked into a flow cell of the 
colorimeter 22 to effect the colorimetric measurement. Next, at the 
position S.sub.20 the carrier 21 contained in the tube 11 is withdrawn via 
the large mouth portion 11a by means of the carrier discharge device 23. 
At the position S.sub.22, the washing liquid shower is intermittently 
supplied into the U-shaped tube 11 by the washing pump 24 and at the same 
time, the washing liquid is discharged through the small mouth portion 11b 
by means of the discharge pump 28. The washing liquid remaining in the 
tube 11 is discharged at the position S.sub.23 by the pump 28. In this 
manner, the U-shaped tube 11 is washed effectively for preparing the 
analysis of another sample. 
Also in the present embodiment, the reaction line is formed as an endless 
line and the U-shaped tubes 11 are repeatedly passed through the washing 
device comprising the washing pump 24 and discharge pump 28 to effect the 
washing including the B-F separation by a plurality of times. Therefore, 
the whole analyzer can be made small, simple and cheap. 
In the above embodiments shown in FIGS. 3, 5 and 6, the number of the 
sample cups 15 held in the sampler 14 is made equal to that of the 
U-shaped tubes 11 supported by the turntable 12, and thus in case of 
analyzing samples the number of which is larger than that of the sample 
cups in the sampler 14, after the completion of the analysis for the 
samples in the sampler 14, a new set of samples must be set in the sampler 
14. Therefore, the operator is subjected to cumbersome and time consuming 
work. 
FIG. 8 is a schematic view showing another embodiment of the 
enzyme-immuno-assay automatic analyzer according to the invention, in 
which the above mentioned drawbacks are obviated. The present embodiment 
differs from the embodiment shown in FIG. 3 only in the carrier supply 
position and the construction of a sampler 41. That is to say, a carrier 
21 is supplied by the carrier supply device 20 into a U-shaped tube 11 at 
a position S.sub.21 between the carrier discharge position S.sub.20 and 
the washing position S.sub.22. Further, the sampler 41 can hold a number 
of racks 43 each supporting a number of sample cups 42 and the racks 43 
are successively transported along a substantially U-shaped path, while 
successive sample cups 42 are indexed at a sample sucking position. 
In the present embodiment, after samples are delivered into all the 
U-shaped tubes 11 held in the turntable 12, the transportation of the 
racks 43 in the sampler 41 is once interrupted. As explained above with 
reference to FIG. 3, during a plurality of rotations of the turntable 12, 
the B-F separation has been effected twice, the test liquid has been 
sucked into the colorimeter 22 to effect the colorimetric measurement, the 
carrier 21 has been removed from the U-shaped tube 11, a new carrier 21 
has been dropped into the U-shaped tube 11, and the washing and delivery 
of buffer solution 26 have been performed. Then, the transportation of the 
racks 43 in the sampler 41 is started again and twenty four samples are 
successively delivered into successive twenty four U-shaped tubes 11 on 
the turntable 12. 
In the present embodiment, it is possible to set a large number of sample 
cups 42 larger than the number of the U-shaped tubes 11 onto the sampler 
41, the labour work of the operator for setting the samples into the 
sampler 41 can be saved materially. Further, since the carrier supply 
position S.sub.21 is set between the carrier discharge position S.sub.20 
and the washing position S.sub.22, the washing of the U-shaped tubes which 
have been used for the analysis of samples can be performed simultaneously 
with the washing of the carriers 21 and thus, the processing speed can be 
increased as compared with the previous embodiments. 
FIG. 9 is a schematic view depicting still another embodiment of the 
enzyme-immuno-assay automatic analyzer according to the invention, in 
which the sandwich method is utilized. In the present embodiment, a number 
of reaction vessels 51 in the form of a test tube are arranged 
equidistantly along an endless belt 52 which is intermittently rotated at 
a given pitch in a vertical plane by means of a pair of driving rollers 
53A and 53B. 
In the present embodiment, a carrier having given antibody or antigen fixed 
thereon is supplied into a reaction tube 51 by means of a carrier supply 
device 54. Next, a given amount of a buffer solution 56 is delivered into 
the tube 51 by a buffer solution delivery device 55. Then, a given amount 
of a sample contained in a sample cup 58 is further delivered into the 
tube 51 by means of a sample delivery device 57 to initiate the first 
antigen-antibody reaction, while the reaction tube 51 is immersed in a 
thermostat 59. The carrier supply device 54, buffer solution delivery 
device 55 and sample delivery device 57 are once kept inoperative after 
the given number of operations corresponding to the number of the reaction 
tubes or samples has been finished. At the end of the thermostat 59, the 
reaction tube 51 and carrier 50 are washed by a washing device 60 to 
effect a first B-F separation. Then the reaction tube 51 is turned 
up-side-down, while the carrier 50 caused to remain in the tube. To this 
end, along the travelling path of the reaction tube there is arranged a 
member 61 for preventing the carrier 50 from being dropped off the 
reaction tube 51. This member 61 may be formed by a mesh. 
During a second revolution of the endless belt 52, a given amount of an 
enzyme-labeled reagent 63 is delivered into the reaction tube 51 by a 
reagent delivery device 62 to start a second antigen-antibody reaction, 
while the tube 51 is transported through the thermostat 59. Then the 
carrier 50 and tube 51 are washed by the washing device 60 to effect a 
second B-F separation. In a third revolution of the endless belt 52, a 
given amount of a color reagent 65 is delivered into the reaction tube 51 
with the aid of a second reagent delivery device 64. After a given 
reaction has been completed, a test liquid in the reaction tube 51 is 
sucked into a colorimeter 66 to effect a colorimetry. After that, the 
reaction tube 51 is washed again by the washing device 60 and the carrier 
50 is dropped off the reaction tube 51 via a swingable gate 68 into a 
waste carrier container 67. 
In the present embodiment, the endless reaction line is formed in the 
vertical plane and a plurality of washings including the B-F separation 
are carried out by means of the single washing device 60 and therefore, 
the whole apparatus can be made small in size, simple in construction and 
cheap in cost. 
FIG. 10 is a schematic view showing still another embodiment of the 
automatic analyzer according to the invention in which three test items 
are analyzed for respective samples by means of the sandwich method. On a 
turntable 71 rotating intermittently in a direction a are arranged 
concentrically three rows of reaction vessels, each including twenty four 
reaction vessels 72. As shown in FIG. 10, the reaction vessels 72 are 
arranged in a radial manner. The successive analyzing steps for respective 
reaction vessels 71 are the same as those explained above in the 
embodiment shown in FIG. 3 and thus will be explained briefly hereinbelow. 
At a position S.sub.17, three carriers are supplied into three reaction 
vessels 72A, 72B and 72C belonging to respective rows by means of carrier 
supply devices 73A, 73B and 73C, respectively. Then at a position 
S.sub.22, the reaction vessels and carriers contained therein are washed 
by a washing device 74 which comprises a wash liquid supplying and sucking 
mechanism. Next, at a position S.sub.24, given amounts of a buffer 
solution 76 are delivered into the three reaction tubes 72A, 72B and 72C 
by a buffer solution delivery device 75. Further, at a position S.sub.1, 
given amounts of a sample in a sample cup 79 in a sampler 78 are delivered 
via sample delivery device 77 into the three reaction vessels 72A, 72B and 
72C to effect a first antigen-antibody reaction. It should be noted that 
given antibody or antigen corresponding to substances in a sample to be 
analyzed is fixed to the carriers, respectively. 
During a second revolution of the turntable 71, the reaction vessels and 
carriers are washed at the position S.sub.22 by the washing device 74 to 
perform a first B-F separation. Then, at a position S.sub.3, 
enzyme-labeled reagents 81A, 81B and 81C corresponding to the respective 
test items are delivered into the reaction tubes 72A, 72B and 72C by means 
of reagent delivery devices 80A, 80B and 80C, respectively to effect a 
second antigen-antibody reaction. In a third rotation of the turntable 71, 
the reaction vessels and carriers are washed by the washing device 74 at 
the position S.sub.22 to perform a second B-F separation. Then, given 
amounts of a color reagent 83 are delivered into the reaction tubes 72A, 
72B and 72C by a second reagent delivery device 82. Next, test liquids in 
the reaction vessels 72A, 72B and 72C are sucked into colorimeters 84A, 
84B and 84C, respectively to effect the colorimetric measurement. Finally, 
at a position S.sub.20 the carriers remaining in the reaction vessels 72A, 
72B and 72C are discharged by a carrier discharge device 85 and then at 
the position S.sub.22 the reaction vessels 72A, 72B and 72C are washed by 
the washing device 74 to prepare the analysis for new samples. 
In the present embodiment, a plurality of test items for respective samples 
can be analyzed simultaneously and thus, the processing ability of the 
analyzer becomes very high. Further, the reaction vessels are repeatedly 
indexed at the washing position S.sub.22 and a plurality of washings 
including the B-F separation can be done by the single washing device 74. 
Therefore, the analyzer becomes small and simple, while the processing 
ability is very high. 
FIG. 11 is a schematic side view depicting an embodiment of the carrier 
supply device for throwing a carrier into a reaction vessel. The carrier 
supply device 91 comprises a hopper 93 for holding a number of carriers 
92. At a lower end of the hopper 93 is connected one end of a passage 94 
having a curved portion and a linear portion. The carriers are rolled down 
into the duct 94 due to the gravitational force and are aligned therein. 
At the other end of the passage 94 is arranged a vertical wall 95 for 
limiting the horizontal movement of carriers and an outlet 96 is formed in 
a lower wall of the duct 94. Near the outlet 96 is rotatably arranged a 
disc 98 in which an opening 97 is formed. By suitably rotating the disc 98 
the carriers 92 can be successively discharged from the duct 94 one by one 
via the outlet 96 and opening 97. The carrier supply device 91 of the 
present embodiment is simple and the carriers 92 can be positively 
supplied into a reaction vessel 99 one by one. 
FIG. 12 is a schematic view showing another embodiment of the automatic 
enzyme-immuno-assay analyzer according to the invention, which utilizes 
the sandwich method shown in FIG. 2. In this embodiment, twenty five 
U-shaped tubes 111 each having a large mouth portion 111a and a small 
mouth portion 111b are used as a reaction vessel as shown in FIGS. 13A to 
13C, and are arranged concentrically on a turntable 112 at a same 
interval. The turntable 112 functions to rotate intermittently the 
U-shaped tubes 111 in the direction of arrow a at a given period for 
example 15 seconds), while the U-shaped tubes are always immersed in a 
thermostat 110 as shown in FIGS. 13A to 13C. Stop positions of the 
U-shaped tube 111 caused by the intermittent rotation of the turntable 112 
are denoted by marks S.sub.1 to S.sub.25. At the stop position S.sub.4, 
the sample to be measured are selectively delivered into the U-shaped tube 
111 from a sample cup 115 positioned at a predetermined sample suction 
position of a sampler 114 by means of a sample delivery device 113. As for 
the sampler 114, various types of samplers can be used. In this 
embodiment, the sampler 114 holds a plurality of racks 114a each having 
ten sample cups. As shown in FIG. 12, the racks 114a arranged in a left 
column in the sampler 114 are successively moved downward to the sample 
delivery position, and the racks 114a arranged in a right column in the 
sampler 114 are moved upward. The rack 114a positioned at the sample 
delivery position is moved intermittently in the direction of arrow S in 
synchronism with the rotation of the turntable 112. When the sample 
delivery operation for all the samples in the rack 114a is finished, this 
rack 114a is moved into the lower end of the right column of the sampler 
114 and then the rack 114a positioned at the lowermost position of the 
left column is moved to the sample delivery position. In this manner, the 
samples to be measured can be successively transferred to the sample 
delivery position at a given pitch. 
At the stop position S.sub.1, a first reagent 117 is selectively delivered 
into the U-shaped tube 111 by means of a first reagent delivery device 
116. As for the first reagent 117, use is made of a buffer solution. At 
the stop position S.sub.3, an enzyme-labeled reagent 119 corresponding to 
the substance to be measured in the sample is selectively delivered into 
the U-shaped tube 111 by means of a second reagent delivery device 118. 
Further, at the stop position S.sub.2, a color reagent 121 is selectively 
delivered into the U-shaped tube 111 by means of a third reagent delivery 
device 120. Furthermore, by utilizing a carrier supply device 122, one of 
insoluble carriers made of a synthetic resin such as plastics or a glass 
bead which are accommodated therein is selectively delivered into the 
U-shaped tube 111 through the large mouth portion 111a. A size of the 
carrier 123 is so determined as to be easily put in and out from the large 
mouth portion 111a and not to be inserted into the small mouth portion 
111b, and to the surface of the carrier 123 is previously fixed an antigen 
or an antibody which causes an antigen-antibody reaction with the 
substance to be measured in the sample. Moreover, at the stop position 
S.sub.20, a reaction liquid supplied in the U-shaped tube 111 is 
selectively sucked into a colorimeter 124, and also at the stop position 
S.sub.23, the carrier 123 accommodated in the U-shaped tube 111 is 
selectively put out by means of a carrier discharge device 125. 
Furthermore, at the stop position S.sub.25, a washing liquid such as an 
ion-exchange water, a buffer solution for an immuno-analysis and a 
physiological saline solution is selectively supplied into and then 
discharged from the U-shaped tube 111 by means of a washing device 126 so 
as to effect the B-F separation and to wash the U-shaped tube 111. 
Hereinafter, an operation of the automatic enzyme-immuno-assay analyzer 
shown in FIG. 12 will be explained with reference to FIGS. 13A to 13C and 
14A to 14I. 
In this embodiment, since use is made of the sandwich method, the analysis 
for each of the samples caused to end within three rotations of the 
turntable 112. That is to say, the B-F separation is effected two times 
and the washing operation of the U-shaped tube is effected once, till the 
end of the analysis for each samples. Therefore, each of the sample 
delivery, the first, second, third reagent delivery, the carrier supply, 
the carrier discharge operations and the sucking operation into the 
colorimeter is effected every three pitches in the course of the rotation 
of the turntable 112. However, since the washing operation is effected 
three times till the end of the analysis for each of the samples as 
mentioned above, it is necessary to effect the washing operation every one 
pitch in the course of the rotation of the turntable 112. Moreover, in 
order to effect the operations mentioned above, it is necessary to arrange 
3n+1 or 3n+2 number of U-shaped tubes 111 on the turntable 112, where n=1, 
2, 3 . . . . In this embodiment, the number of the U-shaped tubes 111 is 
twenty five and thus the requirement of 3n+1 is satisfied at n=8. 
In the course of the first rotation of the turntable 112, at first as shown 
in FIG. 13A, one carrier 123 is supplied into the U-shaped tube 111 
positioned at the stop position S.sub.1 through the large mouth portion 
111a (FIG. 14C). Also at the stop position S.sub.1, a predetermined amount 
of the first reagent 117 consisting of the buffer solution is delivered 
into the U-shaped tube 111 by means of the first reagent delivery device 
116 (FIG. 14D). After this U-shaped tube 111 is transferred by three 
pitches, at the stop position S.sub.4, a predetermined amount of the 
samples is delivered into the U-shaped tube 111 by means of the sample 
delivery device 113 (FIG. 14F). Then, the antigen-antibody reaction is 
initiated from this sample delivery operation. The U-shaped tube 111 
reaches the stop position S.sub.25 at the end of the first rotation, and 
at this stop position S.sub.25 the washing operation for the U-shaped tube 
111 is effected by the washing device 126 so as to perform the first B-F 
separation (FIG. 14B). In FIGS. 14B to 14I, the operational timings for 
the relevant sample are illustrated by hatching. 
Next, in the course of the second rotation of the turntable 112, at the 
stop position S.sub.3 a predetermined amount of the enzyme-labeled reagent 
119 is delivered into the U-shaped tube 111 by means of the second reagent 
delivery device 118 to start the second reaction (FIG. 14F). At the last 
stop position S.sub.25 during the second rotation, the second B-F 
separation is effected by the washing device 126 (FIG. 14B). 
Further, in the course of the third rotation of the turntable 112, at the 
stop position S.sub.2 a predetermined amount of the color reagent 121 is 
delivered into the U-shaped tube 111 by means of the third reagent 
delivery device 120 to start the third reaction (FIG. 14G). Then, at the 
stop position S.sub.20, the test liquid in the U-shaped tube 111 is sucked 
by a pump arranged in the colorimeter 124 and is introduced into a 
colorimetry flow cell to effect the colorimetry thereof with a light 
having a predetermined wavelength (FIG. 14H). Next, at the stop position 
S.sub.23 after three pitch rotations, the carrier 123 remaining in the 
U-shaped tube 111 is removed by means of the carrier discharge device 125 
(FIG. 14I). At the last stop position S.sub.25 during the third rotation, 
the U-shaped tube 111 is washed by the washing device 26 and is used 
repeatedly for the analysis of the next sample. In FIGS. 14C to 14E, the 
operational timings for the next sample to be measured are illustrated by 
cross hatching. 
The washing operation by utilizing the washing device 126 is so performed 
that the washing liquid is intermittently supplied into the U-shaped tube 
111 through the large mouth portion 111a in a shower mode and is 
discharged from the small mouth portion 111b by means of a liquid 
discharge pump. As shown in FIGS. 13A to 13C, the washing device 126 
comprises a washing liquid tank 126a, a washing liquid supply pump 126b, a 
nozzle 126c, a liquid discharge tank 126d and a liquid discharge pump 
126e. Moreover, as shown in FIG. 13A, the carrier supply device 122 
comprises a hopper 122a for accommodating a plurality of carriers 123 and 
a gate device 122b for supplying the carrier 123 one by one from the 
hopper 122a. Generally, the carriers 123 are held in the hopper 122a and 
are wetted with the buffer solution. Further, as shown in FIG. 13C, the 
carrier discharge device 125 functions to descend a nozzle 125a into the 
large mouth portion 111a so as to put out the carrier 123 by sucking it 
through the nozzle, or to descend an arm into the large mouth portion so 
as to put out the carrier by holding it by fingers provided at an end of 
the arm. 
In the embodiment mentioned above, since the sample delivery, the first, 
second, third reagent delivery, the carrier supply and discharge, the 
colorimetry operations are effected every three pitches, and the 
3.times.8+1=25 number of U-shaped tubes 111 are arranged concentrically on 
the turntable 112 with the same interval therebetween, for example at the 
stop position S.sub.4 the U-shaped tube 111 deviates by one pitch every 
one rotation of the turntable 112 viewed in the rotational direction of 
the turntable. Since the same can be applied to the operation which is 
performed every three pitches, the sample delivery operation is performed 
every three pitches. In this manner, the analysis for one sample is 
finished successively every three pitches. Therefore, it is possible to 
effect an ID control of the sample and the processing of the analytical 
result at a constant period, and thus various controls can be effected 
easily. Further, since use is made of an endless reaction line and the 
U-shaped tube 111 is transferred circularly for effecting the washing 
operation including the B-F separation repeatedly by means of one washing 
device 126 arranged in the reaction line, it is possible to make the whole 
apparatus small in size, simple in construction and inexpensive in cost. 
FIG. 15 is a schematic view showing another embodiment of the automatic 
enzyme-immuno-assay analyzer according to the invention, which utilizes 
the competitive method shown in FIG. 1, and FIGS. 16A to 16H are timing 
charts for explaining the operations of the analyzer shown in FIG. 15. 
Portions in FIG. 15 similar to those shown in FIG. 12 are denoted by the 
same reference numerals used in FIG. 12. As explained above with reference 
to FIG. 1, the competitive method needs two washing operations including 
the B-F separation. Therefore, in the embodiment shown in FIG. 15, the 
2n+1 number of the U-shaped tubes 111 are arranged on the turntable 112, 
and the operations are so controlled that the sample delivery, the reagent 
delivery, the carrier supply and discharge, the colorimetry operations are 
effected every two pitches and the washing operation is effected every one 
pitch. In this manner, the sample delivery operation can be successively 
performed every two pitches. 
In the embodiment shown in FIG. 15, at the stop position S.sub.1 the first 
reagent 119 consisting of the enzyme-labeled reagent is delivered into the 
U-shaped tube 111 by means of the first reagent delivery device 118 and 
also one carrier 123 is supplied into the U-shaped tube 111 by means of 
the carrier supply device 122. Moreover, at the stop position S.sub.2 the 
color reagent 121 is delivered into the U-shaped tube 111 by means of the 
second reagent delivery device 120. Further, at the stop position S.sub.3 
the sample is delivered into the U-shaped tube 111 by means of the sample 
delivery device 113. Also in this embodiment, the sample to be measured is 
transferred successively to the sample delivery position by means of the 
sampler 114 which accommodates a plurality of sample racks 114a each 
having a number of sample cups 115. As shown in FIG. 15, the U-shaped tube 
111 which is utilized as the reaction vessel has the large mouth portion 
111a and the small mouth portion 111b. At the stop position S.sub.22, the 
test liquid in the U-shaped tube 111 is sucked into the colorimeter 124 to 
effect the colorimetry. Then, at the stop position S.sub.24, the carrier 
123 remaining in the U-shaped tube 111 is put out by means of the carrier 
discharge device 125, and also at the stop position S.sub.25 the washing 
operation including the B-F separation is performed by means of the 
washing device 126. 
In the timing charts shown in FIGS. 16A to 16H, first of all at the stop 
position S.sub.1 one carrier 123 is supplied into the U-shaped tube 111 by 
means of the carrier supply device 122 as shown in FIG. 16C and also a 
predetermined amount of the enzyme-labeled reagent 119 is delivered into 
the U-shaped tube 111 by means of the first reagent delivery device 118 as 
shown in FIG. 16D. When the U-shaped tube 111 reaches the stop position 
S.sub.3 after two pitch rotations, a predetermined amount of sample is 
delivered into the U-shaped tube 111 to start the antigen-antibody 
reaction as shown in FIG. 16E. Then, at the last stop position S.sub.25 
during the first rotation, the B-F separation is effected by means of the 
washing device 126 as shown in FIG. 16B. After proceeding by two pitches, 
at the stop position S.sub.2 a predetermined amount of the color reagent 
121 is delivered into the U-shaped tube 111 by means of the second reagent 
delivery device 120 to start the second reaction as shown in FIG. 16F. 
When the U-shaped tube 111 reaches the stop position S.sub.22 , the test 
liquid is sucked into the colorimetry flow cell by means of the 
colorimeter 124 to effect the colorimetry thereof as shown in FIG. 16G. 
After proceeding by two pitches, at the stop position S.sub.24 the carrier 
123 remaining in the U-shaped tube 111 is discharged by means of the 
carrier discharge device 125 as shown in FIG. 16H. At the last stop 
position S.sub.25 during the second rotation, the empty U-shaped tube 111 
is washed by the washing device 126 as shown in FIG. 16B to prepare the 
analysis for the next sample. In this manner, predetermined analyses for 
each of the samples can be performed within two rotations of the turntable 
112. Moreover, in this embodiment, since the 2n+1 number of the U-shaped 
tubes 111 are arranged on the turntable 112, and the sample delivery, the 
reagent delivery, the carrier supply and discharge, the colorimetry 
operations are effected every two pitches, successive samples can be 
delivered at two pitch interval. However, the washing operation must be 
effected every one pitch. 
FIG. 17 is a schematic view showing another embodiment of the automatic 
enzyme-immuno-assay analyzer according to the invention, which utilizes 
the sandwich method. Portions in FIG. 17 similar to those shown in FIG. 12 
are denoted by the same reference numerals used in FIG. 12. In this 
embodiment, since, it is necessary to effect the washing operation three 
times during the course of the analysis in case of using repeatedly the 
U-shaped tube, twenty five U-shaped tubes 111 are arranged on the 
turntable 112. The different point between this embodiment and that shown 
in FIG. 12 is that the washing operations are effected simultaneously for 
the three U-shaped tubes 111 positioned at the stop positions S.sub.23 to 
S.sub.25 by means of the washing device 126. In this case, it is not 
necessary to operate the washing device 126 every one pitch, but to 
operate it every three pitches the same as with the other devices. 
Therefore, the driving control is commonly used for all the devices. 
Moreover, in order to achieve such a construction, the carrier discharge 
device 125 is arranged at the stop position S.sub.20. 
FIGS. 18A to 18I are timing charts showing the operations of the analyzer 
shown in FIG. 17. As mentioned above, since the operations are almost 
similar to those shown in FIG. 12 and the different point is that the 
washing operations are effected every three pitches, the explanation of 
these timing charts are omitted here. As clearly understood from the 
timing charts shown in FIGS. 18A to 18I, even in this embodiment, it is 
possible to deliver the samples successively every three pitches. 
FIG. 19 is a schematic view showing another embodiment of the automatic 
enzyme-immuno-assay analyzer according to the invention, which utilizes 
the sandwich method. In the embodiments explained heretofore, one 
concentric reaction line of the U-shaped tubes 111 is arranged on the 
turntable 112, but in this embodiment shown in FIG. 19, three concentric 
reaction lines of reaction tubes 132 each consisting of a test tube are 
arranged on a turntable 131. In this embodiment, each reaction line has 
twenty four reaction tubes 132, but this number of reaction tubes 132 can 
be arbitrarily determined. For the sake of simplicity, outermost, 
intermediate and innermost reaction lines are denoted respectively as a 
first reaction line 132-1, a second reaction line 132-2 and a third 
reaction line 132-3. As with the embodiments mentioned above, the 
turntable 131 is rotated intermittently at a predetermined pitch. A 
carrier supply device 133 is arranged at a stop position S.sub.1 to supply 
the carrier selectively into the reaction tube 132. Moreover, the carrier 
supply device 133 functions to supply the carrier successively into the 
reaction tubes 132 located in the first, second and third reaction lines. 
That is to say, at first the carriers are supplied into all the successive 
reaction tubes 132 arranged in the first reaction line 132-1 one by one, 
and then the carriers are supplied into all the successive reaction tubes 
132 arranged in the second reaction line 132-2 and are finally supplied 
into all the reaction tubes in the third reaction line 132-3. After that, 
this carrier supply operation is effected repeatedly. 
Further, a washing device 134 is arranged at a stop position S.sub.2 to 
wash simultaneously all the reaction tubes 132 located at this stop 
position S.sub.2. A first reagent delivery device 135 is arranged at a 
stop position S.sub.3 to deliver a first reagent 136 consisting of a 
buffer solution into the reaction tube 132. As with the carrier supply 
operation mentioned above, the first reagent 136 is delivered into all the 
reaction tubes arranged in the first, second and third reaction lines, 
respectively in this order. A sample supply device 137 is arranged at a 
stop position S.sub.4 to deliver a sample supplied successively from a 
sample 138 into the reaction tube 132. Also this sample delivery operation 
is performed for the first, second and third reaction lines successively 
in this order. A second reagent delivery device 139 is arranged at a stop 
position S.sub.5 to deliver a second reagent 140 consisting of the 
enzyme-labeled reagent into the reaction tube 132. A third reagent 
delivery device 141 is arranged at a stop position S.sub.6 to deliver a 
third reagent 142 consisting of the color reagent into the reaction tube 
132. Also these second and third reagent delivery operations are performed 
for the first, second and third reaction lines successively in this order. 
Moreover, a colorimeter 143 is arranged at a stop position S.sub.23 to 
effect the colorimetry of the test liquid contained in the reaction tube 
132 after introducing the test liquid into the colorimetry flow cell. 
Further, a carrier discharge device 144 is arranged at a stop position 
S.sub.24 to discharge the carrier remaining in the reaction tube. Also 
these colorimetry and carrier discharge operations are performed for the 
first, second and third reaction lines successively in this order. 
FIGS. 20A to 20I are timing charts for explaining the operations of the 
analyzer shown in FIG. 19, and marks 1, 2, 3 denote the operations for the 
first, second and third reaction lines 132-1, 132-2 and 132-3, 
respectively. As shown in FIG. 20B, only the washing operation is 
performed simultaneously for the first, second and third reaction lines in 
synchronism with the rotation of the turntable 131. 
First of all, at the stop position S.sub.1 the one carrier is supplied into 
the reaction tube 132 arranged in the first reaction line 132-1 as shown 
in FIG. 20C. At the next stop position S.sub.2, this reaction tube 132 is 
washed as shown in FIG. 20B. Since the prior test liquid is caused to 
remain in the reaction tube before the washing operation but this test 
liquid does not include the substance which will be coupled with the 
antibody or antigen on the carrier, the analysis is not affected by a 
contamination therebetween. Further, by washing the U-shaped tube at the 
stop position S.sub.2, it is also possible to eliminate contaminations 
between the previous test liquid and the sample and between the previous 
test liquid and the reagent. At the next stop position S.sub.3, a 
predetermined amount of the first reagent 136 consisting of the buffer 
solution is delivered into the reaction tube by means of the first reagent 
delivery device 135 as shown in FIG. 20D. Then at the stop position 
S.sub.4, a predetermined amount of the sample is delivered into the 
reaction tube by means of the sample delivery device 137 as shown in FIG. 
20E to start the first reaction. In this embodiment, these carrier supply, 
washing, first reagent delivery, sample delivery operations for the 
successive reaction tubes arranged in the first reaction line are 
performed every one pitch. 
When the first reaction tube in the first reaction line into which the 
first sample is delivered reaches again the stop position S.sub.2, the 
first B-F separation is effected by means of the washing device 134 as 
shown in FIG. 20B. In the course of this rotation, the relevant reaction 
vessel passes through the stop positions S.sub.5, S.sub.6, S.sub.23 and 
S.sub.1. However, as clearly seen from FIGS. 20A to 20I, at these stop 
positions the operations are performed for the reaction tubes arranged in 
the second and third reaction lines and thus these operations have no 
objection to the reaction tube in the first reaction line. Then, when the 
reaction tube reaches the stop position S.sub.5, a predetermined amount of 
the second reagent 140 is delivered into the reaction tube by means of the 
second reagent delivery device 139 as shown in FIG. 20F to start the 
second reaction. Then, at the stop position S.sub.2, the reaction tube is 
again washed as shown in FIG. 20B to effect the second B-F separation. 
After that, at the stop position S.sub.6 a predetermined amount of the 
third reagent 142 is delivered into the reaction tube by means of the 
third reagent delivery device 141 as shown in FIG. 20G to start the third 
reaction. When the reaction tube reaches the stop position S.sub.23, the 
colorimetry is performed by means of the colorimeter as shown in FIG. 20H. 
At the next stop position S.sub.24, the carrier remaining in the reaction 
tube is discharged from the reaction tube by means of the carrier 
discharge device 144 as shown in FIG. 20I. Until this point, the reaction 
tube has been rotated three times and the analysis for one sample has been 
finished. In this embodiment, the operations mentioned above are effected 
repeatedly. 
In this embodiment, by operating the carrier supply device 133, at first 
during the first rotation the carriers are supplied one by one into all 
the twenty four reaction tubes arranged in the first reaction line 132-1 
successively, then during the second rotation the carriers are supplied 
one by one into all the twenty four reaction tubes arranged in the second 
reaction line 132-2, and finally during the third rotation the carriers 
are supplied into all the twenty four reaction tubes in the third reaction 
line 132-3. The operations mentioned above are repeatedly effected by the 
carrier supply device 133. As with the operations mentioned above, the 
operations of the sample delivery device 137, the first reagent delivery 
device 135, the second reagent delivery device 139, the third reagent 
delivery device 141, the colorimeter 143 and the carrier discharge device 
144 are effected repeatedly, but the reaction line for which each 
operations is effected is different, as clearly understood from FIGS. 20A 
to 20I. For example, during the first rotation, the carrier supply 
operation is effected to the first reaction line 132-1, while the first 
reagent delivery operation is effected to the first reaction line 132-1 
from the third pitch, the second reagent delivery operation is effected to 
the second reaction line 132-2 till the fourth pitch and to the third 
reaction line 132-3 from the fifth pitch, and the third reagent delivery 
operation is effected to the first reaction line 132-1 till the fifth 
pitch and to the second reaction line 132-2 from the sixth pitch. In this 
manner, according to the present embodiment, since the sample delivery 
operation can be effected every one pitch, it is possible to make the 
sample processing efficiency high as compared with the embodiments 
mentioned previously. Moreover, since the carrier supply and discharge, 
sample delivery, reagent delivery and colorimetry operations are performed 
successively for each of the reaction lines, the control of these 
operations can be made extremely easy. 
In the embodiments of the automatic analyzer so far explained, use is made 
of carriers having given antigen or antibody fixed thereto. Thus there 
must be arranged the device for supplying the carriers into the reaction 
vessels one by one and the device for removing the carriers from the 
reaction vessels. Further, since the antibody or antigen is fixed to the 
carriers having a limited area, an amount of the antigen or antibody fixed 
to the carrier is also limited. Sometimes, it results in a decrease in the 
analysis accuracy. 
According to the invention, the above mentioned drawbacks can be avoided by 
using reaction vessels on an inner wall of which has been fixed given 
antigen or antibody. Now, several embodiments of the automatic analyzer 
using such reaction vessels will be explained. 
FIG. 21 is a schematic view illustrating an embodiment of the automatic 
analyzer according to the invention, in which the enzyme-immuno-assay is 
carried out by the sandwich method. The analyzer comprises a turntable 211 
having thirty seven cuvette holders 213 in which cuvettes 212 can be 
detachably secured. On at least a part of the inner wall of the cuvette 
212 is fixed given antibody or antigen. The turntable 211 is rotated in a 
direction a in a stepwise manner at a given pitch of, for example 15 
seconds. Positions at which the cuvette holders 213 are stopped are 
denoted as S.sub.1 to S.sub.37. In the present embodiment, at a position 
S.sub.1, cuvettes 212 are successively supplied into successive cuvette 
holders 213 of the turntable 211 by means of a cuvette supply device, i.e. 
a cuvette loading device 215. At a next position S.sub.2, the cuvette is 
detected by a detector not shown and when the cuvette held in the cuvette 
holder 213 is detected, the cuvette 212 is washed with a washing liquid by 
means of a washing device 216 to effect the washing and B-F separation. 
At the stop position S.sub.4, a first reagent 218 is delivered by a first 
reagent delivery device 217, at a position S.sub.5 a third reagent 220 is 
delivered by a third reagent delivery device 219, at a position S.sub.6 a 
second reagent 229 is delivered by a second reagent delivery device 221, 
at a position S.sub.7, a sample is delivered by a sample delivery device 
223 from a sample cup 225 situating at a sample sucking position in a 
sampler 224, and at a position S.sub.29 a fourth reagent 227 is delivered 
by a fourth reagent delivery device 226. As the first, second, third and 
fourth reagents 218, 229, 220 and 227, use is made of a buffer solution, 
an enzyme-labeled reagent, a color reagent, i.e. enzyme substrate, and a 
reaction stop reagent for stopping the reaction between the enzyme-labeled 
reagent and color reagent. In the present embodiment, the sampler 224 
holds a number of racks 224a each supporting ten sample cups 225. The 
racks in a left column are successively moved downward in FIG. 21 and are 
then shifted leftward as shown by an arrow S into the sample sucking 
position. After all the ten samples in a rack have been delivered into 
cuvettes 212, the rack 224a is moved rightward into a lower end of a right 
rack column. Then the right column of racks is moved upward. In this 
manner a large number of samples in the sampler 224 can be successively 
indexed at the sample sucking position. 
At a stop position S.sub.32, a test liquid contained in the cuvette 212 is 
measured by a colorimeter 228. After that, at a position S.sub.35 the 
cuvette 212 is removed from the turntable 211 by means of a cuvette 
discharge device 222. In the present embodiment, the test liquid is 
subjected to the direct photometry, while the test liquid is caused to 
remain in the cuvette and then the cuvette 212 and the test liquid are 
separately collected into a waste cuvette container and a waste liquid 
container, respectively. 
Now the operation of the analyzer shown in FIG. 21 will be described also 
with reference to FIGS. 22A to 22C and 23A to 23J. 
In the present embodiment, respective samples are analyzed by the sandwich 
method during a period in which the turntable 211 is rotated by about 
three revolutions. Therefore, the delivery of the first to fourth 
reagents, the supply and discharge of cuvettes and the colorimetry are 
effected once each time the turntable 211 is rotated by three pitches. 
However, since the washing is effected by three times for respective 
cuvettes 212, the washing device 216 is operated each time the turntable 
211 is rotated by one pitch. To this end, at the position S.sub.2 there is 
provided the detector for detecting the existence or non-existence of the 
cuvette. Further, in order to operate the analyzer in this manner, the 
number of cuvettes 212 held in the cuvette holders 213 must be set to 3n+1 
or 3n+2, wherein n is a positive regular number 1, 2, 3 . . . . In the 
present embodiment, n=12 and there are provided thirty seven cuvette 
holders 213. 
During a first revolution of the turntable 211, at the position S.sub.1, a 
cuvette 212 is supplied into a cuvette holder 213 by the cuvette supply 
device 215 as shown in FIG. 22A. On the inner wall of the cuvette there 
has been previously fixed given antibody or antigen. At the position 
S.sub.2, the cuvette 212 is washed by the washing device 216. At the 
position S.sub.4 which is downstream with respect to the position S.sub.1 
by three pitches, a given amount of the first reagent 218, i.e. the buffer 
solution, is poured into the cuvette 212 by the first reagent delivery 
device 217 to start the first antigen-antibody reaction. In FIGS. 23B to 
23J various operations effected for the relevant cuvette are denoted by 
hatching. 
During a second revolution of the turntable 211, at the position S.sub.2 
the first B-F separation is performed by the washing device 216. Then, at 
the position S.sub.6 a given amount of the second reagent 229, i.e. the 
enzyme-labeled reagent, is delivered into the cuvette 212 by means of the 
second reagent delivery device 221 to initiate the second antigen-antibody 
reaction. 
During a third revolution of the turntable 211, at the position S.sub.2 the 
second B-F separation is carried out by the washing device 216. Next at 
the position S.sub.5 a given amount of the third reagent 220, i.e. the 
color reagent containing the enzyme substrate, is delivered into the 
cuvette 212 to start the color reaction. Then, at the position S'.sub.22 a 
given amount of the reaction stop reagent 227 is poured into the cuvette 
by the fourth reagent delivery device 226. At the position S.sub.32, the 
test liquid formed in the relevant cuvette 212 is directly measured by the 
colorimeter 228 via the cuvette. At the position S.sub.35, the cuvette 212 
is discharged from the turntable 211 by the cuvette discharge device 229. 
After the turntable 211 has been rotated by three pitches, a new cuvette 
is delivered into the relevant cuvette holder by the cuvette supply device 
215. In FIGS. 23A to 23E, the operations for this newly supplied cuvette 
are denoted by cross hatching. 
As explained above, the analysis for a single sample is carried out for a 
time period during which the turntable 211 is rotated by substantially 
three revolutions. In the present embodiment, the sample delivery, the 
first to fourth reagent deliveries, the cuvette supply and discharge and 
colorimetry are effected every time the turntable 211 is moved by three 
pitches and further the number of cuvettes held in the turntable is set to 
3.times.12+1=37. Therefore, cuvettes which are successively indexed at, 
for instance the position S.sub.7 deviate one by one every time the 
turntable is rotated by one revolution. This is the case for all the 
operations which are effected once every time the turntable is rotated by 
three pitches. Therefore, the sample delivery can be performed regularly 
once for every three pitches and thus, the ID control for samples and the 
treatment of analytic data can be effected at a constant period. In this 
manner, various kinds of control can be carried out simply. Also in this 
embodiment, a plurality of washings including the B-F separation can be 
performed by passing the cuvette repeatedly through the single washing 
device and therefore, the analyzer can be made small in size and simple in 
construction. Moreover, since the cuvette is used as the carrier for 
fixing given antigen or antibody, it is not necessary to use the separate 
carriers in the previous embodiments and thus, the running cost can be 
decreased. 
FIG. 24 is a schematic view illustrating another embodiment of the 
automatic analyzer according to the invention, in which the 
enzyme-immuno-assay is performed in competitive method by using the 
cuvettes having given antibody or antigen fixed on inner walls thereof. In 
the present embodiment, portions similar to those of the embodiment shown 
in FIG. 21 are denoted by the same reference numerals used in FIG. 21. In 
the competitive method, the washing is effected twice and thus, 2n+1 
cuvette holders 213 are provided in the turntable 211. Then, the sample 
delivery, reagent delivery, supply and discharge of cuvette and 
colorimetry are performed once every time the turntable is rotated by two 
pitches. At a position S.sub.3, the cuvette is detected by a detector (not 
shown) and the washing is effected by the washing device 216. At a 
position S.sub.5, a given amount of a first reagent, 222, i.e. the 
enzyme-labeled reagent is poured into the cuvette by the first reagent 
delivery device 221 and at the same time a given amount of a sample is 
poured by the sample delivery device 223. Also in the present embodiment, 
successive samples contained in sample cups 225 are indexed into the 
sample sucking position by means of the sampler 224 comprising a number of 
sample cup racks 224a. At a position S.sub.6, a given amount of a second 
reagent 220, i.e. the color reagent, is delivered into the cuvette by the 
second reagent delivery device 219. At a position S.sub.32, a given amount 
of a third reagent 227, i.e. the reaction stop reagent, is poured by the 
third reagent delivery device 226. Further, at a position S.sub.34, the 
test liquid contained in the cuvette is measured by a colorimeter 228 via 
the cuvette and at a position S.sub.36, the waste cuvette 212 is removed 
from the cuvette holder 213 by the cuvette discharge device 229 and the 
vacant cuvette and test liquid are contained in separate containers. 
FIGS. 25A to 25I show timing charts of various operations of the analyzer 
illustrated in FIG. 24. At the position S.sub.1, a cuvette 212 is supplied 
into a cuvette holder 213 of the turntable 211. After the rotation of the 
turntable by two pitches, at the position S.sub.3 the cuvette is washed by 
the washing device 216. After further two pitches, at the position S.sub.5 
the enzyme-labeled reagent 222 and sample are delivered into the cuvette 
212 to initiate the antigen-antibody reaction. During the second 
revolution of the turntable 211, the B-F separation is effected at the 
position S.sub.3 by the washing device 216. Then, the color reagent 220 
is poured into the cuvette 212 by the delivery device 219 to initiate the 
color reaction. This reaction is finished when the reaction stop reagent 
227 is supplied into the cuvette by means of the delivery device 226. 
Next, at the position S.sub.34, the test liquid is measured by the 
colorimeter 228 via the cuvette 212. Finally, at the position S.sub.36, 
the cuvette is removed from the turntable 211 by the cuvette discharge 
device 229. In FIGS. 25B to 25I, the operations relating to the relevant 
sample are denoted by hatching and operations for a next sample are 
represented by cross hatching. In this manner, the analysis for respective 
samples can be performed every time the turntable is rotated by two 
revolutions. Further, the various operations such as the sample delivery, 
reagent deliveries, supply and discharge of cuvette and colorimetry are 
performed once each time the turntable is rotated by two pitches and thus, 
the control of the analyzer can be effected in a simple manner. It should 
be noted that the washing operation by the washing device 216 has to be 
performed every time the turntable 211 is rotated by one pitch. 
Now concrete constructions of the various portions of the automatic 
analyzers shown in FIGS. 21 and 24 will be explained. 
FIG. 26A is a perspective view showing an embodiment of the cuvette type 
reaction vessel according to the invention and FIGS. 26B and 26C are cross 
sections cut along lines I--I and II--II in FIG. 26A, respectively. In the 
present embodiment, the cuvette 212 is formed by a mold of transparent 
synthetic resin and has generally a flat box shape. The cuvette 212 has an 
opening 212a at its top, two main walls 212b and 212c, two side walls 212d 
and 212c and a bottom wall 212f. Onto at least a part of the inner wall of 
the cuvette is fixed given antibody or antigen which selectively reacts 
with substance to be analyzed. In one of the main walls 212b there is 
formed a T-shaped projection 212g which serves to hold the cuvette in a 
cuvette holder due to its elastic force as will be explained later. The 
side walls 212d and 212e of the cuvette are arranged perpendicularly to a 
measuring optical axis and include entrance and exit windows 212h and 212i 
for a measuring light beam. As best shown in FIG. 26B, the windows 212h 
and 212i are retarded inwardly with respect to the side walls 212d and 
212e, respectively and side edges of the main walls. That is to say, the 
measuring windows 212h and 212i are surrounded by the main walls 212b and 
212c and the bottom wall 212 f and therefore, the windows are effectively 
protected against stain and injury, which ensures a high measuring 
accuracy. Further, an inner surface of the bottom wall 212f is formed 
semi-cylindrically. This results in that the photometry can be carried out 
for a very small amount of the test liquid. 
The antibody or antigen can be fixed to the inner surface of the cuvette 
made of plastics by means of the known physical absorption method or 
chemical binding method. Further, if the existence of proteins such 
antigen, antibody and labeling enzyme might influence the photometry, the 
windows 212h and 212i are kept free from the antibody or antigen. 
FIG. 27 shows an embodiment of a photometering station in which an 
absorbance of the test liquid contained in the cuvette 212 is measured. A 
light beam emitted from a lamp 228a is collimated by a lens 228b and is 
made incident upon the entrance window 212h of the cuvette 212 via a stop 
228c. A light flux emanating from the exit window 212i is made incident 
upon a light detector 228f by means of a stop 228d and an optical filter 
228e. The cuvette 212 is held in a recess 213a of the cuvette holder 213 
formed in the turntable 211 at its periphery due to the elasticity of the 
cuvette. 
A large number of cuvettes 212 are arranged in a magazine 230 as 
illustrated in FIG. 28. It is not necessary at all for an operator of the 
analyzer to insert the cuvettes in the magazine, but the magazine having 
the cuvettes previously contained therein is available. Therefore, the 
cuvettes can be further protected against the stain and injury. The 
magazine 230 may be formed by a mold of plastics or metal. In the present 
embodiment, the magazine has such a length viewed in a direction A that 
ten cuvettes are arranged side by side and such a width measured in a 
direction B that also ten cuvettes are arranged side by side. Therefore, 
the magazine can contain a hundred cuvettes in a matrix form. In a side 
wall 231a of the magazine 230 is formed an outlet 231b having a width 
which is substantially equal to the width of the cuvette 212 and a height 
which is nearly equal to the height of the cuvette 212. In order to ensure 
that the cuvette 212 can be discharged out of the magazine 230 through the 
outlet 231b in a correct posture, a resilient strip 231c is formed in a 
front wall of the magazine at a portion adjacent to the outlet 231b by 
providing a recess in the wall. In top walls 231d and bottom walls of the 
magazine are formed three recesses 231f. It should be noted that the 
recesses 231f do not extend in the upper wall 231d up to the front edge so 
that the cuvettes in a first column are not situated under the recesses. 
This ensures smooth movement of the cuvettes. In the magazine there is 
inserted a push plate 232 between the assembly of the cuvettes and the 
rear wall 231e. As will be explained hereinafter, the cuvette array may be 
moved in the direction B by moving the plate 232 in this direction B. 
Along a right hand side edge of the side wall 231a is formed a step 231g 
which avoids an inverse insertion of the magazine into an automatic 
cuvette loader having a corresponding projection. 
FIGS. 29 and 30 illustrate an embodiment of the automatic cuvette loader 
according to the invention for supplying the cuvettes 212 in the magazine 
230 one by one into successive recesses 213a of the cuvette holder 213 of 
the turntable 211. The turntable 211 is rotated in a direction a in FIG. 
29 in a stepwise manner at the given pitch to form a circular reaction 
line. 
The cuvette auto-loader supply device, i.e. a cuvette supply device, 
comprises a base plate 240, and to a rear surface of the base plate 240 is 
secured a magazine container 241 as best shown in FIG. 30. In the magazine 
container 241 is arranged movably up and down a magazine support 242 to 
which is secured one end of a wire 244 whose other end is connected via a 
pulley 243 to a weight 246 which is movably supported in a cylindrical 
guide 245. Therefore, the magazine support 242 is biased upwardly. In the 
magazine container 241 there may be arranged a plurality of the magazines 
230 each containing a hundred cuvettes 212. Pusher 283 is urged against 
the upper surface of magazine 230 as shown and coil spring 284 biases 
pusher 283 against magazine 230. 
In the base plate 240 is formed a first opening 240a above the magazine 
container 241 and the opening 240a has such a dimension that the magazine 
230 can pass therethrough. On the upper surface of the base plate 240 are 
arranged L-shaped levers 248 and 249 rotatable about shafts 248a and 249a, 
respectively. To these levers are secured ring-shaped stoppers 250 and 251 
by means of shafts 250a and 251a, respectively. As explained later, to the 
levers 248 and 249 are also secured rollers 252 and 253 by means of shafts 
252a and 253a. In free ends of the L-shaped levers 248 and 249 are further 
formed projections 248b and 249b, respectively. Besides the opening 240a 
of the base plate 240 there are further arranged L-shaped posts 254 and 
255 and stoppers 254a and 255a are secured to the posts. 
In the base plate 240 there is further formed a second opening 240b through 
which the magazine can be passed. Beside the second opening 240b a pair of 
magazine support levers 256 and 257 are arranged rotatably about shafts 
256a and 257a. Near free ends of these levers are secured pins 256b and 
257b, respectively, these pins being engaged with the projections 248b and 
249b of the levers 248 and 249, respectively. To the levers 256 and 257 
are further secured pins 256c and 257c which are engaged with projections 
of push levers 258 and 259 which are arranged rotatably about shafts 258a 
and 259a, respectively extending in parallel with the plane of the drawing 
of FIG. 29. The levers 248, 249, 256, 257, 258 and 259 are biased by means 
of springs not shown into positions shown by solid lines. When the levers 
256 and 257 are moved as depicted by imaginary lines, the push levers 258 
and 259 are rotated in a plane perpendicular to the plane of the drawing 
of FIG. 29. 
To the base plate 240 are further secured two guide shafts 260a and 260b by 
means of leg portions 261a and 261b, the guide shafts extending above the 
first and second openings 240a and 240b. A first slider 262 is movably 
secured to the guide shafts 260a and 260b by means of linear bearings. To 
the first slider 262 is secured a wire 263 which is wound around a pulley 
264 provided on the leg portion 261a, a pulley 266 secured to a driving 
shaft of a motor 265 and a pulley 267 provided on the leg portion 261b. 
When the motor 265 is driven in both directions, it is possible to move 
the first slider 262 in a direction B along the guide shafts 260a and 260b 
in a reciprocal manner. By this movement, it is possible to transfer the 
magazine 230 situated above the first opening 240a into a cuvette charging 
position above the second opening 240b and to feed the cuvettes 212 in the 
magazine 230 in the direction B. For this purpose, to the lower surface of 
slider 262 are secured three arms 262a which can be inserted in the 
recesses 231f formed in the magazine 230 and are made in contact with the 
push plate 232. 
There are further provided a pair of guide shafts 268a and 268b extending 
perpendicularly to the guide shafts 260a and 260b, the guide shafts 268a 
and 268b being coupled with the base plate 240 by means of leg portions 
269a and 269b. To these guide shafts 268a and 268b is slidably mounted a 
second slider 270 to which is connected a wire 271 extending around a 
pulley 272 secured to the leg portion 269a, a pulley 274 connected to a 
driving shaft of a motor 273 and a pulley 274 connected to a driving shaft 
of a motor 273 and a pulley 275 secured to the leg portion 269b. When the 
motor 273 is driven in both directions, the second slider 270 can be moved 
reciprocally in the direction A along the guide shafts 268a and 268b. By 
this movement, the cuvettes 212 in the magazine 230 can be inserted into 
the recesses 213a of the cuvette holder 213 one by one. To this end, to 
the slider 270 is slidably secured a pin 270a to which a pushing claw 270c 
is connected and a coiled spring 270b is arranged around the pin 270a. 
Then, it is possible to push resiliently the cuvette 212 situated at an 
extreme position in the magazine 230 by means of the pushing claw 270c. 
There is inserted a coiled spring 276 between the guide shaft 268b and the 
leg portion 269a and the guide shaft 268b is slidably mounted on the leg 
portion 269a and therefore, the guide shafts 268a and 268b are biased in 
the leftward direction in FIG. 29. Further, as shown in FIG. 30, the 
slider 270 is slidably secured to the guide shaft 268a by means of a 
linear bearing, but is coupled with the guide shaft in a frictional manner 
by means of a coiled spring 277 and a ball 278. Therefore, the slider 270 
and guide shaft 268b can be moved together over a certain limited range. 
To the other end of guide shaft 268b is connected an L-shaped rail 
supporting member 279 to which is secured a guide rail 280 having a trough 
construction whose width is substantially equal to the width of the 
cuvette 212, The guide rail 280 is supported by guide rollers 281a, 281b, 
281c and to 281d and can be moved in the direction A over a relatively 
small distance. Near a tip of the guide rail is arranged a leaf spring 282 
for pressing the cuvette situated at the tip of guide rail 280. 
Now the operation of the automatic cuvette loader of this embodiment will 
be explained. It is assumed that in the magazine container 241 there are 
set several magazines 230 and the uppermost magazine is engaged with the 
stoppers 250 and 251 secured to the levers 248 and 249, respectively, so 
that the magazine stock does not move upwardly furthermore. Above the 
second opening 240b is positioned a magazine 230 which is supported by the 
levers 256 and 257, so that it does not fall down in the opening 240b. The 
magazine situated at the cuvette charging position above the second 
opening 240b contains a number of cuvettes 212 to be successively supplied 
into the respective recesses 213a of the cuvette holder 213 of the 
turntable 211. When the motor 273 is driven in a forward direction, the 
wire 271 is rotated in the clockwise direction in FIG. 29 and thus, the 
slider 270 is moved in the direction A. At the same time, the guide shaft 
268b is moved also in the direction A and thus, the rail supporting member 
279 and guide rail 280 are also moved in the direction A. During this 
movement, the first cuvette column (the uppermost horizontally aligned 
cuvettes in FIG. 29) is moved also in the direction A. The guide shaft 
268b is moved in the direction A until a nut 268c provided on the right 
hand end of guide shaft 268b is engaged with the leg portion 269a, and 
after that, only the slider 270 is further moved in the direction A. By 
this movement of the slider 270, the cuvette column is moved in the 
direction A and the left hand cuvette is discharged out of the guide rail 
280 and is inserted into the recess 213a of the cuvette holder 213, which 
recess is just situated opposite to the guide rail 280. As explained 
above, the cuvette 212 has the T-shaped projection 212g formed in its 
main wall 212b and therefore, the cuvette 212 is resiliently clamped in 
the recess 212a. Next the motor 173 is driven in the reverse direction and 
the slider 270 and guide shaft 268b are moved in a direction opposite to 
the direction A until the rail supporting member 279 makes contact with 
the leg portion 269b. During this reverse movement, the pushing claw 270c 
of the slider 270 is always caused to make contact with the cuvette. 
By repeating the above operation, successive cuvettes 212 in the uppermost 
column in FIG. 29 can be supplied into respective recesses 213a of the 
cuvette holder 213 one by one. After that the motor 273 is driven in the 
reverse direction and the slider 270 is returned into the right hand 
position in FIG. 29. Then, the motor 265 is driven in a forward direction 
by a predetermined amount and the wire 263 is rotated in the 
counter-clockwise direction in FIG. 29. During this movement, the first 
slider 262 is moved in the direction B by a distance equal to the width of 
the cuvette 212 and the cuvettes remaining in the magazine 230 are moved 
in the direction B by means of the pushing plate 232. 
In this manner, all the cuvettes 212 in the magazine 230 can be 
successively supplied into the reaction line constituted by the cuvette 
holder 213 in the turntable 211. After that, the motor 265 is driven in a 
reverse direction and the slider 262 is moved in a direction opposite to 
the direction B into the lowermost position in FIG. 29. At the end of this 
movement of the slider 262, the slider 262 is engaged with the rollers 252 
and 253 on the levers 248 and 249. Then the levers 248 and 249 are rotated 
into positions shown by chain lines and the ring-shaped stoppers 250 and 
251 are disengaged from the magazine 230. Then the uppermost magazine in 
the magazine container 241 is moved above the base plate 240 via the first 
opening 240a and is engaged with the stoppers 254a and 255a. When the 
levers 248 and 249 are rotated, the arms 256 and 257 are also rotated by 
means of the engagement of the projections 248b and 249b with the pins 
256b and 257b, into positions illustrated by chain lines, so that the 
empty magazine is fallen down through the second opening 240b. In order to 
enhance the operation for discharging the empty magazine from the cuvette 
loading position, the pushing levers 258 and 259 are rotated in 
conjunction with the rotation of the arms 256 and 257 so as to push the 
magazine downward. 
Finally, the motor 265 is driven in the forward direction and the slider 
262 is moved upward in FIG. 29 and the levers 248, 249, 256, 257, 258 and 
259 are returned to the positions shown by solid lines. In this manner, 
the new magazine can be set into the cuvette loading position. 
The present invention is not limited to the embodiments explained above, 
but may be modified in various manners. For instance, in the above 
embodiments a hundred cuvettes are contained in a single magazine, but any 
desired number of cuvettes may be contained in the magazine. Further, a 
single array of a plurality of cuvettes may be arranged in an elongated 
magazine. In such a case, it is not necessary to move the cuvettes in the 
direction B in the magazine. Moreover, in the above embodiment, the used 
cuvettes are wasted, but they may be used repeatedly after washing them. 
It should be further noted that the construction of the cuvette is not 
limited to that shown in FIGS. 26A to 26C. 
For instance, the T-shaped projection 212g may be formed in both the main 
walls 212b and 212c. Further, legs may be provided by extending the side 
walls 212d and 212e beyond the bottom wall 212f. In this case, the outer 
bottom surface may be also shaped circularly corresponding to the 
semi-cylindrical inner bottom surface. 
According to the embodiment just explained above, the reaction cuvettes can 
be successively supplied into the reaction line in a positive and accurate 
manner without staining and injuring the cuvettes, so that the reliability 
and accuracy of measurement can be increased materially. Further, the 
automatic cuvette loader has a simple construction and can be made less 
expensive. Moreover, the entrance and exit windows of the cuvette are 
effectively protected by the surrounding walls against stain, injury and 
stray light, and the measuring precision can be made much higher. Further, 
a number of cuvettes are contained in the magazine and thus, and the 
transportation and management of the cuvettes can be easily effected 
without staining and injuring the cuvettes. Moreover, the cuvettes may be 
supplemented in a prompt manner during the analysis without interrupting 
the measurement. 
FIG. 31 is a schematic view showing another embodiment of the 
enzyme-immuno-assay automatic analyzer according to the invention which 
performs the sandwich method explained above with reference to FIG. 2. On 
a turntable 312 are arranged equidistantly twenty four U-shaped tubes 311 
along a periphery of the turntable. The turntable 312 is intermittently 
rotated in a direction shown by an arrow a at a given period of, for 
example 15 seconds, while the U-shaped tubes 311 are dipped into a 
thermostat. Positions at which the U-shaped tubes 311 are stopped due to 
the stepwise rotation of the turntable 312 are denoted as S.sub.1 to 
S.sub.24. In the present embodiment, into a U-shaped tube 311 positioned 
at S.sub.1 is delivered a sample from a sample cup 315 which is situated 
just at a sample sucking position of a sampler 314 by means of a sample 
delivery device 313. The sampler 314 holds twenty four sample cups 315 
arranged equidistantly along a disc which is rotated intermittently in a 
direction b in synchronism with the rotation of the turntable 312. Into a 
U-shaped reaction tube 311 situated at S.sub.17 is supplied a carrier 312 
such as a synthetic resin particle or glass bead from a carrier supply 
device 320. It should be noted that the carrier 321 has a diameter smaller 
than an inner diameter of the large mouth portion 311a of the U-shaped 
tube 311, but is larger than an inner diameter of the small mouth portion 
311b. On an outer surface of the carrier 321 there has been previously 
fixed antibody or antigen which causes the antigen-antibody reaction with 
antigen or antibody substance in the sample to be tested. Further, in the 
carrier supply device 320, the carriers 321 are wetted with a buffer 
solution. A reaction liquid in a U-shaped tube 311 at a position S.sub.19 
is selectively sucked into a colorimeter 322, and a carrier 321 contained 
in a U-shaped tube 311 at a position S.sub.20 is removed therefrom by 
means of a carrier discharge device 323. Into a U-shaped tube 311 at a 
position S.sub.22 is supplied a washing liquid such as ion exchange water, 
buffer solution for immunological analysis, physiological saline solution, 
etc. by means of a washing pump 324. At position S.sub.2, a stirring air 
pump 327 can be detachably connected to small mouth portions 311b of 
U-shaped tubes 311, and at positions S.sub.22 and S.sub.23, a discharge 
pump 328 can be detachably connected to small mouth portions 311b of 
U-shaped tubes 311. In a U-shaped tube situating at a position at S.sub.24 
is selectively delivered one of a buffer solution 330, an enzyme-labeled 
reagent 331 and a color reagent 332 by means of a reagent delivery device 
329 of a syringe type. 
On end of a reaction tube is connected to the reagent delivery device 329, 
and the other end thereof is connected to a nozzle 335 which is 
transferred to a position S.sub.24 or a washing liquid tank 333 by means 
of a transfer means not shown. Moreover, a valve 334 is arranged in a 
middle of the reagent tube, to which a buffer solution tank 330, an 
enzyme-labeled reagent tank 331 and a color reagent tank are connected 
through respective tubes. By changing the valve 334, the buffer solution, 
the enzyme-labeled reagent and the color reagent are selectively delivered 
into the U-shaped tube 311 positioned at the position S.sub.24. 
Now, the operation of the automatic analyzer shown in FIG. 31 will be 
explained. 
During a first revolution of the turntable 312, at the position S.sub.17, a 
carrier 321 wetted with the buffer solution is supplied in a U-shaped tube 
311 via its large mouth portion 311a. Then, at the position S.sub.22, the 
washing liquid is intermittently poured into the U-shaped tube 311 from 
the large mouth portion 311a by means of the washing pump 324 and at the 
same time, the washing liquid is sucked out of the tube 311 via the small 
mouth portion 311b by means of the discharge pump 328. Next, at the 
position S.sub.23, any washing liquid remaining in the tube 311 is 
discharged by the discharge pump 328. 
Then, at the position S.sub.24 a given amount of the buffer solution is 
delivered into the U-shaped tube 311 via its large mouth portion 311a by 
means of the delivery device 329 after changing the valve 334 to select 
the buffer solution tank 330. Then, at the position S.sub.1 a given amount 
of a sample is delivered by means of the sample delivery device 313 into 
the tube 311 from a sample cup 315 situated at the sample sucking position 
of the sampler 314. Next, at the position S.sub.2, an air stream is 
supplied into the U-shaped tube 311 by means of the air pump 327 to stir 
the buffer solution and sample in the tube 311. In this manner, a first 
antigen-antibody reaction is initiated. It should be noted that the 
carrier supply device 320, buffer solution delivery device 325, sample 
delivery device 313 and sampler 314 are made inoperative after being once 
operated for respective U-shaped tubes. 
During a second revolution of the turntable 312, at the position S.sub.22, 
the liquid in the tube 311 is sucked via the small mouth portion 311b by 
the discharge pump 328 and at the same time, the washing liquid is 
intermittently poured into the tube via its large mouth portion 311a by 
means of the washing pump 324. The washing liquid remaining in the tube is 
discharged at the positions S.sub.22 and S.sub.23. In this manner, the 
U-shaped tube 311 and the carrier 321 contained therein are fully washed 
to effect a first B-F separation. Then, at the position S.sub.24, a given 
amount of the emzyme-labeled reagent is delivered into the U-shaped tube 
311 via its large mouth portion 311a by the reagent delivery device 329 
after charging the valve 334 to select the enzyme-labeled reagent tank 
331. The reagent and carrier are stirred sufficiently at the position 
S.sub.2 by supplying the air stream from the small mouth portion 311b with 
the aid of the air pump 327 to effect a second antigen-antibody reaction. 
During a third revolution of the turntable 312, at the position S.sub.22, 
the U-shaped tube 311 and carrier are washed by means of the washing pump 
324 and discharge pump 328 to perform a second B-F separation. Next, a 
given amount of the color reagent is delivered into the U-shaped tube 311 
by the reagent delivery device 329 after changing the valve 334 to select 
the color reagent tank 332. Then, the color reagent and carrier are 
stirred by means of the air pump 327 to start a reaction of the color 
reagent with the labeling enzyme of the enzyme-labeled reagent bound with 
the carrier 321. 
In a fourth revolution of the turntable 312, at the position S.sub.19 a 
reaction liquid in the U-shaped tube 311 is sucked into the colorimeter 
322 to effect the colorimetric measurement. 
At the position S.sub.20, the carrier 321 is sucked out of the U-shaped 
tube 311 via its large mouth portion 311a by the carrier discharge device 
323. At the position S.sub.22, the washing liquid is supplied into the 
U-shaped tube 311 via its large mouth portion 311a by means of the washing 
pump 324 and the wash liquid is sucked out of the tube by means of the 
discharge pump 328. The wash liquid remaining in the tube is discharged at 
the position S.sub.23 by means of the discharge pump 328. In this manner, 
the U-shaped tube 311 is prepared for a next supply of a carrier. During 
the operations mentioned above, the nozzle 335 is transferred to the 
washing liquid tank 333 after the end of each reagent delivery operation 
so as to wash inner and outer walls of the nozzle 335 by means of the 
syringe 329. 
The present invention is not limited to the embodiments explained above, 
but many modifications and alterations can be conceived, which are within 
the scope of the invention. In the above embodiments, the enzyme-labeled 
reagent is used to perform the enzyme-immuno-assay, but the 
radio-immuno-assay and flourescent-immuno-assay may be also adopted. 
Further, it is not always necessary to use a circular reaction line, but 
the reaction line can be formed by a snake chain. Moreover, in the 
embodiments using the carriers, the direct colorimetry may be effected, 
while the test liquid is caused to remain in the reaction vessel. In this 
case, if the carrier affects the measurement, it may be withdrawn from the 
reaction vessel prior to the colorimetry. Moreover, in all the above 
embodiments there is provided the single washing device, but a plurality 
of washing devices may be used. For instance, in the embodiment shown in 
FIG. 3, two washing devices may be arranged at diametrically opposite 
positions with the first washing and second B-F separating being performed 
by the first washing device and the first B-F separation and last washing 
being carried out by the second washing device. Even in such a case, the 
number of the washing devices can be decreased as compared with the case 
in which the four washing operations are carried out by four separate 
washing devices. Further, in the embodiments using the carriers, the 
reaction vessels may be discharged. 
In the embodiments explained above, a single test item is measured by a 
single reaction line, but the multiple test items may be analyzed by a 
single reaction line. Further, in order to improve the reliability of 
analysis, the same test item may be measured by two reaction lines. 
Moreover, the deliveries of the reagents and buffer solution are carried 
out at different positions, but they may be effected at the same position. 
Then, the agitation may be performed only at a single position. Further, 
in the embodiment shown in FIG. 5, the delivery of the buffer solution 
prior to the carrier supply may be performed by the delivery device 25, 
and then the separate buffer solution delivery device 31 may be delected. 
In the embodiments illustrated in FIGS. 21 and 24, the cuvette is washed 
after the cuvette supply, but prior to the sample delivery. However, since 
the cuvette is discharged from the turntable after analysis, saic 
preliminary washing may be dispensed with. Further, the test liquid may be 
colorimetered by introducing the test liquid from the cuvette into a flow 
cell of a separate colorimeter. Then, the delivery of the reaction stop 
liquid may be omitted. Further, in the embodiments, use is made of the 
cuvette of flat box shape, but cuvette may be formed in any desired shape. 
Moreover, in order to effect the reaction more stably, the cuvette may be 
immersed into a thermostat in the reaction line.