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
  • G01N21/75—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated
  • G01N21/77—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator
  • G01N21/82—Systems in which material is subjected to a chemical reaction, the progress or the result of the reaction being investigated by observing the effect on a chemical indicator producing a precipitate or turbidity

Definitions

• the present invention is concerned with a method for the measurement of the properties of an agglutination, a precipitate, or of a corresponding reaction result placed 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 concerned with an apparatus for the implementation of the method, which apparatus comprises one or several detectors receiving radiation, the said detectors being located so that the measurement beam received by them passes substantially in the vertical direction of the vessel, as well as an output unit.
• 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 measurement Saovaniemi, Osmo, "Performance and Properties the Finnpipette Analyzer System", Proceedings of the Second National Meeting on Biophysics and Biotechnology in Finland, 183, 1976
• one beam of light does not produce a suf ficient difference in absorbance between agglutination a non-agglutination.
• any uncertainty is eliminated by performing the measurement of the properties of the agglutination precipitate at several points once or several times so as to observe the formation of the precipitate as a function of time.
• the agglutinated precipitate formed on the bottom of the reaction vessel is, viz., structurally different from a non-agglutinated precipitate.
• the forme 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, in order to measure the formation, location, and form of the precipitate and/or the density or other properties of different points of the precipitat a component field of limited area, out of the field of the vessel bottom to be measured, is measured; the vessel and/or the source of radiation and/or the detector is/are moved so that the component field subject to measurement moves along the field to be measured; and the measurement result of the component field at each particular time under measurement is read constantly or at specified intervals for the purpose of processing and output of the information.
• the apparatus in accordance with the invention is characterized in that, at one time, either the source of radiation or the detector receiving radiation covers only a limited component field of the bottom of the vessel and that the vessel and/or the source of radiation and/or the detector receiving the radiation can be shifted so that the location of the component field to be measured out of the bottom of the vessel at the bottom of the vessel is changed.
• an either slit-shaped or subtaritially point-shaped source of light is placed underneath the bottom of the reaction vessel, in its immediate proximity, which source of light illumi nates only a little part of the precipitate at a time.
• the reaction vessel and the source of light are shifted in relation to each other, the light passes alt ⁇ rnatingly through different parts of the precipitate, whereby it is possible to measure and to register the light absorption and/or transmittance at each particular point of the precipitate altematingly by means of the detector placed above the reaction vessel. Since an agglutinated precipitate gives a line of light absorbance and/or transmittance values different from that given by a non-agglutinated precipitate, it can be decided easily electronically which case is concerned.
• reaction vessel is shifted laterally so that the beam of measurement alternatingly meets different points of the precipitate placed on the bottom of the vessel, or
• the beam of measurement is shifted laterally so that it meets different points of the precipitate placed in a stationary reaction vessel alternatingly.
• the movements described in sections 2 and 3 can be performed in more than one directions, whereby, when a point-shaped source of light or detector is used, the topography of the precipitate placed on the bottom of the vessel can be examined completely.
• the principles described in sections 1 to 4 can be applied to a multi-channel apparatus, such as, e.g. the "FP-9" photometer. 6.
• the principles described in sections 1 to 4 can also be applied to apparatuses measuring phenomena other than light absorption or transmittance (fluorometer, luminescence, etc.).
• Figures 1a and 1b illustrate two different cases in a reaction vessel
• Figure 2 schematically illustrates an alternative in which one slit-shaped source of light is used that can be shifted in relation to the reaction vessel.
• Figure 3 shows an absorbance curve obtained from the reaction vessel of Fig. 2; underneath the curve, the reaction vessel of Fig. 2 is shown as viewed from the top (in the curves of the figures, the x-axis illustrates the location of the point of measurement on the path of movement and the y-axis illustrates the absorbance at each particular time or in each particular point),
• Figure 4 shows absorbance curves obtained by means of the source of light shown in Fig. 5 ; underneath the curves the precipitate measured is shown as viewed from above, and
• Figure 5 is a schematical presentation of a source of light that consists of several point-shaped sources of light.
• Figure la there is a precipitate 4a on the bottom of the reaction vessel 2, the shape and density of the said precipitate being at different points different from those of the precipitate 4b shown in Figure 1b.
• the precipitate 4a illustrates an agglutinated situation
• figure 4b represents a non-agglutinated precipitate in the situation of measurement.
• Figure 2 schematically shows a slit-shaped source of light 1, a reaction vessel 2, and a detector 3.
• the length of the slit is preferably somewhat less than the diameter of the vessel bottom.
• the absorbance curve 3 shown in Fig. 3 is obtained, which curve may be continuous or consist of individual points.
• Figure 3 also shows a top view of the precipitate 4 on the bottom 2 of the reaction vessel. The measurement has been performed by at a time always measuring a stripe of the shape of the slit-shaped source of light out of the precipitate en the bottom of the reaction vessel and by producing .the output of the measurement value of this stripe.
• Figure 4 shows a precipitate 4 measured from the bottom of the reaction vessel 2 so that the reaction vessel has passed across the sources of light 8 to 10 arranged in a line and shown in Fig. 5, so that measurement has first been performed by means of the source of light 8 and the other sources of light 9 and 10 have been closed. Then the reaction vessel has come back, and then the measurement has been performed by means of the beam 9 whereas the beams 8 and 10 have been closed.
• the beams 8 and 9 have been closed and the measurement has been performed by means of the beam 10, when the reaction vessel has by-passed it.
• the beam 10 is corresponded by the scan 13 and by the absorbance curve 16.
• the beams 9 and 8 correspond the scans 12 and 11 and the absorbance curves 15 and 14.
• the reaction vessel moves back and forth at its measurement point a number of times equalling the number of measurement beams in operation. It is also natural that one dot-shaped source of light may move along the slit shown in Figure 5 , being alternatingly in positions 8, 9 and 10.
• the source of light and the detector may change places, whereby the detector is placed immediately underneath the reaction vessel and it is given a slitshaped or dot-shaped form.
• the method of measurement may be based on photometry 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 turbidometry, fluorometry, or, e.g., on the use of a source of radiation and a receiver for luminescence, 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. It is natural that, in stead of one method of measurement, the reaction vessels may be measured simultaneously or subsequently by means of two or more wave lengths or methods of measurement (e.g., photometry and fluorometry), the final result being based on the infor mation thereby obtained.
• the reaction vessels may be measured simultaneously or subsequently by means of two or more wave lengths or methods of measurement (e.g., photometry and fluorometry), the final result being based on the infor mation thereby obtained.
• the sources of light arranged in a line shown in Figure 5
• the said detectors moving in relation to the reaction vessel.
• the detectors may be in operation either all of them at the same time, in which case one scan movement is adequate, or each of them alternatingly, whereby the scanning is performed correspondingly several times.
• Movement of the field of measurement in relation to the bottom of the vessel can be arranged either by shifting the vessel or the dot-shaped or slit-shaped source of light or the detector, or both.

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• 1981
  • 1981-07-24 EP EP19810902325 patent/EP0056413A1/de not_active Withdrawn
  • 1981-07-24 WO PCT/FI1981/000057 patent/WO1982000354A1/en not_active Ceased

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