RU2398233C2 - Device for estimating concentration of immuno-active objects in samples of biological liquids - Google Patents

Abstract

FIELD: medicine.

SUBSTANCE: device contains precipitation plate with applied on it layer of gel, which contains antibodies, illuminator of precipitation plate in form of source of diffusive light of toroidal form and reading videodevice, connected with computer. Precipitation plate is made in form of disk and placed on base, provided with light adsorbing coating from precipitation plate side, in such way, that geometrical centers of reading videodevice, illuminator and precipitation plate are placed on one vertical axis.

EFFECT: device application allows to provide even contrast range of immuno-diffusive rings image on precipitation plate, to read them with required accuracy, and increase productivity of measurement process.

3 cl, 4 dwg

Description

The invention relates to the field of biomedical laboratory equipment and can be used in immune studies of various biological objects.

A device for determining the concentration of antibodies in samples of biological fluids containing a precipitation plate, a lighting system for a precipitation plate, a lighting system for labels for measuring the diameters of immunodiffusion rings on a precipitation plate, a unit for observing alignment of labels with the boundaries of the immunodiffusion ring, and a digital display for displaying measurement results, US 4099881 , cl. G01B 11/02, 1978.

This device is based on the use of the interactive operator-device principle, in which the measurement process is reduced to the manual control of sequential supply of labels from ring to ring when observing the process with an eye. The known device has inherent disadvantages due to the constant participation of the operator; the device cannot be effectively used to measure biological objects due to its low productivity and the high probability of errors due to subjective factors. The lighting scheme does not allow creating contrast images of immunodiffusion rings on the entire plate, as a result of which the device can hardly be automated, since due to different contrasts, the results will be accompanied by both false positive and false negative distortions.

A device is also known for determining the concentration of antibodies in samples of biological fluids, including a precipitation plate onto which a gel containing antibodies is applied, a precipitation plate illuminator in the form of directional light sources, a reader with a mask, a two-dimensional scanning device for a precipitation plate in the field of view of a video device, and also a personal computer for controlling a video device with a mask, a two-dimensional scanning device and statistical processing of the results and measuring diameters of immunodiffusion rings, DE 4318692, cl. G01N 33/559, 1994

In the known device, illumination is produced by a system that does not allow uniform illumination of the immunodiffusion rings on the entire precipitation plate, especially around the periphery, because, due to different refractive indices, the gel-glass system behaves like a fiber; in this case, the manifestations of edge effects are most significant and lead to different contrast images of the rings on the precipitation plate. In addition, in the known device a very negative effect on the contrast of the images of the rings is the fact that the radiation source has a directivity. At small thicknesses of the photometric object (gel thickness is 1 mm and the thickness of the transparent substrate is about 0.5 mm), the absorption of directional light is small, which sharply reduces the contrast of the image, in particular, immunodiffusion rings.

In connection with the foregoing, the device does not allow without distortion to simultaneously read images of all immunodiffusion rings and, accordingly, their simultaneous measurement; but because the measurement process is quite lengthy and inefficient.

The closest known to the claimed invention is an analyzer of immunoactive objects (Exhibition: World of Biotechnology-2005, Russian Academy of Sciences - Medicine and Human Health, March 14-18-18, 2005, http://www.expo.ras.ru/base/prod_data. asp? prod_di = 1847), which includes a precipitation plate with a layer of gel containing antibodies on it, a precipitation plate illuminator in the form of a toroidal diffuse light source and a video reading device connected to a computer.

This device has two significant drawbacks due to the geometry of the optical circuit. Firstly, this system cannot provide sufficient uniformity of illumination due to the manifestation of edge effects leading to different contrast images of the rings on the precipitation plate. Secondly, the absence of a light-absorbing substrate does not allow to obtain the maximum possible contrast of the image of the rings.

Thus, the prototype device does not allow efficiently and without distortion to read images of all immunodiffusion rings.

The objective of the present invention is to provide uniform high contrast images of all immunodiffusion rings on a precipitation plate and thereby the possibility of their simultaneous reading with the required accuracy; the result is an increase in the productivity of the measurement process.

According to the invention, in a device for determining the concentration of immunoactive objects in samples of biological fluids, including a precipitation plate with a layer of gel containing antibodies on it, a precipitation plate illuminator in the form of a toroidal diffuse light source and a video reading device connected to a computer, the precipitation plate is made in the form disk and mounted on a base provided with a light-absorbing coating on the side of the precipitation plate, while the centers of the video reader, illuminator and precipitation plate are located on the same vertical axis; the diameter of the precipitation plate cannot exceed the inner diameter of the illuminator; the distance d from the horizontal plane of symmetry of the illuminator to the upper surface of the precipitation plate can be determined from the relation:

,

,

Where

t is the radius of the cross section of the illuminator,

r is the distance between the center of the cross section of the illuminator and the vertical axis of symmetry of the illuminator.

A hood can be installed between the illuminator and the video reader.

The applicant has not identified any technical solutions identical to the claimed one, which allows us to conclude that it meets the criterion of "novelty."

Thanks to the implementation of the distinguishing features of the invention, the object acquires important new properties consisting in providing uniform illumination and obtaining the maximum image contrast of all immunodiffusion rings on the entire precipitation plate, which allows simultaneously to read images of all immunodiffusion rings without distortion and to measure them simultaneously. As a result, the accuracy and speed of measurements made using the device are increased.

The above allows, according to the applicant, to conclude that the claimed technical solution meets the criterion of "inventive step".

The invention is illustrated by drawings, which depict:

figure 1 - diagram of the device, a longitudinal section;

figure 2 is a diagram explaining the relative position of the illuminator and the precipitation plate;

figure 3 - precipitation plate with immunodiffusion rings, top view, on an enlarged scale;

figure 4 is a section a - a in figure 3.

A device for determining the concentration of immunoactive objects in samples of biological fluids includes a precipitation plate 1 mounted on the base 2. Plate 1 in a specific example is made of transparent polystyrene (GOST 20282-86). Base 2 is made of duralumin grade D-16. The lighter 3 is made in the form of a toroidal diffuse light source, in particular, an LC 22W ring fluorescent lamp, OSRAM, Germany, with a luminous flux of 1350 lm is used. The video reader 4 is a high-sensitivity digital camera Matrix 1400 M manufactured by Deltatekh LLC, Moscow. Video device 4 is connected to a computer (not shown conventionally in the drawings). In this example, we used a personal computer with a Pentium IV processor, 2.0 gigahertz, RAM - 1 gigabyte.

The precipitation plate 1 is made in the form of a disk. The base 2 from the side of the plate 1 is equipped with a light-absorbing coating 5 made of cotton black velvet fabric 1 mm thick.

The geometric centers of the video reader 4, the illuminator 3 and the precipitation plate 1 are located on the same vertical axis 6.

The diameter of the plate 1 (2R) does not exceed the inner diameter (D) of the illuminator 3; the distance d from the horizontal plane 7 of symmetry of the illuminator 3 to the upper surface of the plate 1 is determined from the ratio:

,

,

Where

t is the radius of the cross section of the illuminator,

r is the distance between the center of the cross section of the illuminator and the vertical axis of symmetry of the illuminator.

This ensures maximum brightness of the plate 1 with its required uniformity.

A hood 8 is installed between the illuminator 3 and the video reader 4, which prevents direct light from the illuminator 3 from directly entering the lens of the video device 4.

The device operates as follows.

A uniform gel layer 9 of a gel, in particular agar, 1 mm thick is applied to the surface of the precipitation plate 1. The gel contains antibodies specific for the studied immunoactive object. Through the gel layer 9, through holes in the form of holes 10 are made at predetermined points using a vacuum punch. In a series of holes in the Central zone 11 of the precipitation plate 1 make a calibration fluid with increasing values of the concentration of the studied immunoactive object in a given range.

In the remaining wells make different samples (taken from different sources - people, animals) of biological fluids: blood serum, lymph, saliva, cerebrospinal fluid, etc.

As a result of the interaction of immunoactive objects of biological fluids with gel antibodies, diffusion rings 12 of various diameters are formed, the value of which is proportional to the concentration of the immunoactive object in this sample of biological fluid.

Diffuse light passes from the illuminator 3 through the gel layer 9 and the precipitation plate 1 and is partially absorbed. The part of the light that passed through the gel layer 9 and the precipitation plate 1 is absorbed by the light-absorbing coating, and the part of the light that fell on the diffusion rings 12 is scattered, forming contrast images of the diffusion rings 12. These images are read by the video device 4 and transmitted to a computer in digital form, where their diameter is determined and compared with the diameters of the rings formed when the precipitation plate 1 of the calibration fluid is introduced into the wells of zone 11.

The results of this comparison determine the concentration of immunoactive objects in all samples of biological fluids.

For the manufacture of the device used conventional structural materials and equipment. This circumstance allows, according to the applicant, to conclude that the present invention meets the criterion of "industrial applicability".

Claims (3)

1. A device for determining the concentration of immunoactive objects in samples of biological fluids, including a precipitation plate with a layer of gel containing antibodies on it, a precipitation plate illuminator in the form of a toroidal diffuse light source and a video reading device connected to a computer, characterized in that the precipitation plate made in the form of a disk and mounted on a base equipped with a light-absorbing coating on the side of the precipitation plate, while The centers of the video reader, illuminator and precipitation plate are located on the same vertical axis. 2. The device according to claim 1, characterized in that the diameter of the precipitation plate does not exceed the inner diameter of the illuminator, while the distance d from the horizontal plane of symmetry of the illuminator to the upper surface of the precipitation plate is determined from the ratio:

,
where t is the radius of the cross section of the illuminator,
r is the distance between the center of the cross section of the illuminator and the vertical axis of symmetry of the illuminator. 3. The device according to claim 1, characterized in that a hood is installed between the illuminator and the video reader.

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