Classifications

• • G—PHYSICS
  • G01—MEASURING; TESTING
  • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
  • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
  • G01N21/17—Systems in which incident light is modified in accordance with the properties of the material investigated
  • G01N21/25—Colour; Spectral properties, i.e. comparison of effect of material on the light at two or more different wavelengths or wavelength bands
  • G01N21/251—Colorimeters; Construction thereof
  • G01N21/253—Colorimeters; Construction thereof for batch operation, i.e. multisample apparatus

Definitions

• the present invention is concerned with a metho for the measurement of the properties of an agglutination, a precipitate, or of a corresponding reaction result place on the bottom of a vessel by means of radiation and of a detector that receives radiation, whereat the beam of measurement coming from the source of radiation passes substantially in the direction of the vertical axis of the vessel, and the intensity of the radiation passing through, or reflected from, the precipitate on the bottom of the vessel is measured.
• the invention is also con ⁇ cerned with an apparatus for the implementation of the method, which apparatus comprises a detector receiving radiation, the said detector being located so that the measurement beam received by it passes substantially in the direction of the vertical axis of the vessel, an output unit, and possibly a source of radiation.
• the objective of the method in accordance with the invention is to be able to ascertain the difference between agglutination and non-agglutination sufficiently clearly, reproducibly, and carefully.
• the principle of vertical mea ⁇ surement Saovaniemi, Os o, "Performance and Properties of the Finnpipette Analyzer System", Proceedings of the Second National Meeting on Biophysics and Biotechnology in Finland, 183, 1976
• the agglutinated precipitate formed on the bottom of the reaction vessel is, viz., structurally different from a non-agglutinated precipitate.
• the former is, e.g., unhomogeneous, at the middle part denser than at the sides, whereas the latter is even and relatively homogeneous.
• the method in accordance with the invention is characterized in that the measurement of the formation, location, and form of the precipitate placed on the bottom of the vessel and/or of the density or other properties of different points of the precipitate is performed in respect of component fields of limited area separately, -the measurement result being read for each
• the apparatus in accordance with the invention is charac ⁇ terized in that the detector consists of several sub- detectors placed in the same plane close to one another.
• the employment of a detector matrix may take place, e.g., in the following two ways:
• the performance of the measurement at several points is performed by employing a detector matrix composed of small sub-detectors.
• the detector matrix can be positioned directly underneath a reaction vessel which is open at the top or placed under a trans ⁇ parent cover, whereby the reaction vessel is illuminated from above by means of homogeneous light.
• Each sub- detector registers the light absorption of the portion of the precipitate placed directly above the said sub- detector, and the electronics of the apparatus processes the results and decides whether the precipitate is agglu ⁇ tinated or not.
• a source or radiation built in the apparatus or it is possible to use the daylight, the general illumination of the room, or any other source of light that gives uniform light. Stability of the source of light is in this case not critically necessary, because the evaluation of the result is simultaneously based on a comparison of the signals received from the different sub-detectors, which comparison can be performed electronically.
• a detector matrix consisting of several sub-detectors is used for each reaction vessel, which sub-detectors of the matrix register the light-absorbing or light-emitting property of the precipitate or any other reaction result on the bottom of the vessel only from a certain point, each sub-detector from an individual point of its own.
• the precipitate to be examined is illumi ⁇ nated appropriately, e.g., by means of homogeneous light. It is possible to use an external, sufficiently homo ⁇ geneous light coming primarily from above, such as, e.g., daylight, the general illumination of the room, or any other available source of radiation.
• the signals received from the sub-detectors are compared with each other and/or with blank, reference, and/or standard values, e.g., electronically, and it is decided whether the result equals, e.g., agglutination or not.
• the apparatus may be provided with a permanently programmed output unit, which directly indi ⁇ cates whether the precipitate in the vessel is aggluti ⁇ nated or not, e.g. as + and - display. . 6)
• the apparatus may be a multi-channel appa ⁇ ratus, in which case it has a detector matrix of its own for each channel.
• the channels may be arranged as a matrix corresponding the extensively used and widely spread, e.g., so-called pit plates, cuvette sets, etc.
• Figures la and lb illustrate two different cases in a reaction vessel
• Figure 2 is a schematical presentation of one embodiment of the apparatus in accordance with the invention
• Figure 3 illustrates absorbance readings obtai ⁇ ned by means of the apparatus shown in Fig. 2, and under ⁇ neath the graphical presentation the measured precipitate is shown as viewed from above
• Figure 4 is a schematical presentation of a second embodiment of the apparatus in accordance with the invention.
• Figure 5 is a partly sectional view of the apparatus in accordance with the invention, wherein it is possible to measure the precipitates of several vessels simultaneously.
• the precipitate 3 ⁇ a illustrates an agglutinated situation
• Figure 34b represents a non-agglutinated precipi ⁇ tate in the situation of measurement.
• Figure 2 shows a reaction vessel 32 open at the top, which reaction vessel contains the reaction mixture as well as the precipitate 3 on the bottom 33.
• sub-detectors 36 are seen which register the light coming from the source of radiation 37 and passing through the precipitate 34.
• the sub-detectors are arranged in a line and/or in a matrix consisting of rows. It is evident that, in stead of the square sub-detector matrix shown in the figure, it is also possible to use matrixes of other shapes, such as, e.g., of the shape of a line, circle, etc., and it is possible to apply detectors consisting, e.g., of sectors and rings placed one inside the other.
• Figure 3 shows how, out of the measurement arrangement shown in Fig. 2, each sub-detector (1 to 25) yields the corresponding absorbance values (1 to 25).
• the x-axis of the system of coordinates illustrates the location of the line to which the sub-detector belongs, and the y-axis illustrates the absorbance.
• Figure 4 shows a reaction vessel 32, which contains the reaction mixture 31 and, on the bottom, the precipitate 34, and a source of light 37 underneath the vessel. Above the vessel there is a matrix detector 35 and between the vessel and the detector a lens 38. By means of the lens 38, an exact image of the precipitate 34 is formed on the matrix detector 35_, whereby each sub- detector 36 receives light in accordance with what the precipitate 34 has allowed to pass through at its different points.
• the quantity of radiation falling onto each detector from the incoming radiation depends on the absorption, transmission, scattering, or any other pheno ⁇ menon of the precipitate 34 at the different points of the precipitate.
• Figure 5 shows how it is possible, for example, to construct an apparatus in accordance with this prin ⁇ ciple wherein it is possible simultaneously to measure the precipitates occurring in all the reaction vessels of a so-called " microtiter-disk. Underneath each reaction vessel 32 there is a detector matrix 35 of its own. An electronic computer unit built in the construction of the apparatus decides whether the precipitate is agglutinated (+) or not (-) and indicates the result on the monitor screen 39.
• the method of measurement may be based on photo- metry or multiphotometry, the latter meaning a photometer which comprises several channels so that each sample has a source of light and a detector of its own.
• the method of measurement may, of course, being a single-channel or multi-channel method, be additionally based, e.g., on turbidometr , fluorometry, or, e.g., on the use of a source of radiation and a receiver for lumi ⁇ nescence, laser beam, ultrasound, etc. phenomena.
• the positioning of the reaction vessels or equivalent, of sources of measurement beams, of detectors, etc. auxiliary equipment may be performed in the way most appropriate in each particular case.
• the equipment may also involve various degrees of automation, e.g., in the pipetting of the samples and reagents, in the shifting of the beams of measurement, and in the processing of the results.
• reaction vessels may be measured simul- taneously or subsequently by means of t ⁇ o or more wave lengths or methods of measurement (e.g., photometry and luorometry), the final result being based on the infor ⁇ mation thereby obtained.
• t ⁇ o or more wave lengths or methods of measurement e.g., photometry and luorometry
• a source of radiation it is possible to use either a source of radiation placed in the apparatus, a source of radiation placed apart from the apparatus, or the general illumination of the room, e.g. the daylight.

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• Physics & Mathematics (AREA)
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• 1981
  • 1981-07-24 EP EP19810902327 patent/EP0056414A1/de not_active Withdrawn
  • 1981-07-24 WO PCT/FI1981/000059 patent/WO1982000355A1/en not_active Ceased

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