FR2589239A1 - Agglutinographic reaction chamber - Google Patents

Abstract

The invention relates to agglutinographic reaction chamber for reactive liquid particles. According to the invention, it comprises, in combination, a first transparent panel 21 having a first surface of a first predetermined area, a second panel 20 having a second surface of a second predetermined area, the second surface being coextensive with and overlapping at least part of the first surface, the second panel being spaced apart by a predetermined distance from the first so that the overlapping parts of the first and second surfaces define a capillary chamber 15 in order to draw the liquid reactants between the overlapping parts of the first and second surfaces when a reactive liquid is introduced to or around the capillary chamber. The invention applies in particular to reactions of immunochemical particles.

Description

 The present invention is generally directed to the reaction of reactive immunochemical particles and in particular to an agglutinographic reaction chamber which causes, intrinsically, the agglutination of the reactive immunochemical particles and presents a stable and high contrast visual recording of the reaction. 'agglutination. The process by which stable, high-contrast visual recording of an agglutination reaction of immunochemical particles occurs, without the need for external movement, is referred to herein as "agglutinography" or "agglutinographic reaction".

 Methods for the reaction of reactive immunochemical particles using laboratory coverslips are well known but show results which are far from satisfactory. These known methods are usually characterized by placing liquid reagents on a glass coverslip and light rocking and swirling of the coverslip to force the liquid reactants to swirl and form agglutinations. Without the swirling of the reactive particles, there is no reliable formation of visible agglutinations.

 Typically, to obtain optimum performance, the swaying and swirling of the coverslip must be accomplished in an exact manner, for a specific period of time. At the end of the swinging and swirling time, the results should be read immediately. This known method produces incorrect results in several conditions. If the swirling is carried out too quickly or too slowly or if the duration of the swinging and swirling is too short or too long, the reagents either do not react completely or react too much. If the reaction results are not observed at the end of the swing period, it may continue to react.

Conventional agglutination reagents also evaporate within a few minutes, quickly destroying the visual record of the particulate reaction.

If the reagents protrude beyond the edges of the coverslip during rocking or other handling, this will naturally result in imprecise results.

 An agglutinographic reaction chamber for reactive liquid particles is provided. The reaction chamber consists of a first panel and a second panel covering at least a portion of the first panel. The non-overlapping area of the first panel defines a reception region. The first panel is separated from the second panel at a predetermined distance to define a chamber between said first and second panels. The introduction of liquid reactive particles to or around the receiving region causes a capillary force which displaces the liquid reactants in the chamber and forces an agglutination reaction to occur between the first and second panels.

 The object of the present invention is to provide an agglutinographic reaction chamber.

 Another object of the present invention is to provide an agglutinographic reaction chamber which causes the formation of agglutinations without swaying or swirling.

 Another subject of the present invention is an agglutinographic reaction chamber which causes the formation of agglutinations which are reproducible and easy to observe without the need to delay the reaction.

 Another object of the invention is to provide an agglutinographic reaction chamber which produces a recording of the agglutinographic reaction.

 Another object of the present invention is to produce an agglutinographic reaction chamber which is simple to use and produces reproducible agglutination reactions.

The invention will be better understood, and other objects, characteristics, details and advantages thereof will appear more clearly during the explanatory description which follows, made with reference to the appended schematic drawings given solely by way of example illustrating several embodiments of the invention, and in which
- Figure 1A is a plan view of an agglutinographic reaction chamber constructed according to a mode
preferred embodiment of the invention;
- Figure 1B is a side elevational view of the agglutinographic reaction chamber of Figure 1A;
- Figure 1C is a partially enlarged view of region C surrounded by a circle in Figure lB ;;
- Figures 2A-2C are progressive cross-sectional views of an agglutinographic reaction occurring in the agglutinographic reaction chamber of Figure 1A;
- Figures 3A-3C are progressive plan views of the agglutinographic reaction respectively shown in Figures 2A-2C;
- Figure 4 shows another embodiment of the agglutinographic reaction chamber constructed according to the invention;
- Figure 5 is another embodiment of an agglutinographic reaction chamber constructed according to the invention;
- Figure GA is a plan view of another embodiment of an agglutinographic reaction chamber constructed according to the invention;
- Figure 6B is a side elevational view of the agglutinographic reaction chamber of Figure 6A;
- Figure 7 is a side elevational view of a
another embodiment of a reaction chamber
agglutinographic constructed according to the invention;
- Figure 8A is a plan view of another mode
of an agglutinographic reaction chamber
built according to the invention; and
- Figure BB is a side elevation view
of the agglutinographic reaction chamber of the figure
8A.

We will first refer to Figures lA-1C where a
agglutinographic reaction chamber, generally -indxquee - at 10, and and constructed according to the invention, is shown. The agglutinotraphic reaction chamber
10 consists of a glass top panel 20 and a
glass bottom panel 21 with a narrow space
DD defined between the panels 20, 21 delimited by.

spacers 30 to produce a capillary chamber
plane 15. A reception region 22 is defined by the
surface to which the smallest panel 20 does not cover
not the panel 21. Immunochemical reagent particles in a liquid 23 are applied to the receiving region 22. Reagents 22 are placed near the end 26 of the capillary chamber 15. Particles & tV-esiiufflnocni-miques 2- 3 are then drawn into the
capillary chamber 15 by capillary action and are forced
to be drawn across the whole room. The signs
20, 21 can be of equal size and the reagents
23 are then introduced at an entrance to the room
capillary 15.

In order to understand how a reaction test
agglutination occurs using the agglutination reaction chamber of the present invention, the
The following basic steps explain an agglutination test.

A test sample, a reactive antibody and a latex
reactive polystyrene would be pipetted onto the
coverslip 10 to reception region 22.

The reagents and the test sample 23 are drawn through the capillary chamber 15. If the test sample is absent from the molecule of interest, the reactive sample begins to react and to form small agglutinations. This capillary flow, which is intrinsic to the agglutinographic reaction chamber, produces the driving force for the formation of agglutinations. As soon as two particles react to form an agglutination, their combined speed is reduced compared to the unreacted particles
This speed reduction is predicted by Stoke's law
F = 6 nrv; v = F x 1
6 nr Stoke's law indicates for a particle in a liquid of viscosity n, the speed v of the particle relative to the speed of the liquid is inversely proportional to the radius r of the particle.

 As a result, agglutination of particles moves more slowly than non-agglutinated particles. Likewise, larger agglutinations of particles move more slowly than smaller agglutinations of particles. Furthermore, in the case where the molecule of interest is present, the agglutination reaction takes place in proportion to the quantity of the molecule of interest present.

 Referring to Figures 1A-1C, while liquid reactants move from right to left, unreacted particles which are behind agglutination collide with agglutination due to the relative speed difference which exists between unreacted particles and agglutinated particles. These collisions result in even larger agglutinations which move even more slowly and are thus exposed to an ever increasing rate of collision with unreacted particles and smaller agglutinations.

 This reactive process continues until the size of the agglutinations increases to the point where their speed approaches zero, at which point any unreacted particle either sticks to the agglutination or continues beyond the agglutinations in the surrounding liquid flow. the agglutinated particles. The result of this situation is a very high-contrast visual presentation of the agglutinations when they occur in the test sample.

 Reference will now be made to FIGS. 2A-2C and 3A-3C, which shows a sequential illustration of an agglutinographic reaction occurring in the agglutinographic reaction chamber of FIG. 1.

FIGS. 2R and 3A represent the condition of the reaction while the agglutinographic reagents 23 are deposited, with a pipette, on the receiving region 22 of the panel 21 near the position 26 of the capillary chamber 15. The reagent 22 is attracted by capillary action in the capillary chamber 15. The liquid reagents 23 will be continuously drawn into the capillary chamber 15 in the direction of the arrow 27 until the far end of the capillary chamber 15 is reached.

 Reference will now be made to FIGS. 2B and 38 where liquid reagents 23 have entered the capillary chamber 15 and have started to agglutinate in small agglutinations 26. While the liquid reagents 23 are drawn into the capillary chamber 15, a liquid flow in the direction of the arrow 27 forces the agglutinographic particles in the reagent 23 to start to agglutinate into small agglutinations 28.

 We will now refer to FIGS. 2C and 3Ç where a complete agglutinographic reaction is represented.

The reaction ends when the capillary forces stop forcing the reagents to flow and when the agglutinographic reaction chamber visually stabilizes.

 As can be more clearly seen in Figure 3C, there is a very high degree of agglutination uniformly visible across the entire surface of the capillary chamber.

 As the agglutinographic reaction is maintained between two glass panels 20, 21, the liquid does not evaporate quickly and the whole of the reaction remains stable. As a result, one can see for much longer than one could in the agglutinographic reaction of the prior art, in the open air.

 In a preferred embodiment of the invention shown in Figure 1, an agglutinographic reaction chamber 10 is constructed as follows. The lower panel 21 and the upper panel 20 are made of glass. The lower panel 21 is for example a glass strip of 50 mm x 50 mm by 1 mm and the upper panel 20 is a glass strip of 50 mm x 40 mm x 1 mm. These two glass panels are separated by a distance of approximately 3 μm to create a capillary chamber 15. Reagent 23 consists of a mixture of agglutination reagents such as a 0.3 m diameter polystyrene latex coated with HCG and an HCG antibody in solution.

 The panels 20 and 21 need not of course have these dimensions or necessarily be made of glass. However, at least one of the lower panel 21 and the upper panel 20 must have a wettable surface forming a surface of the capillary chamber 15.

 The distance DD between the upper sheet 20 and the lower sheet 21 can vary from approximately 0.1-500 km, it is preferably 2-20 μm and better still 3-7. The optimal distance DD depends on the dimension of unreacted particles, The larger the unreacted particles, the greater the distance DD must be for an optimal agglutinographic reaction to occur.

Likewise, smaller unreacted particles react more efficiently in the capillary chamber 15 having a smaller distance D-D.

 The distance D-D is maintained between the upper panel 20 and the lower panel 21 par. spacers 30. A large number of different types of spacers can be used to maintain the spaced relationship between the sheets 20, 21. The spacers 30 can be either painted, on screen, silk, ink, polyester film, dust, surface irregularities on the slats or a separate film. Various materials will give the best results depending on the distance D-D to be used in a particular agglutinographic reaction chamber, the reagents to be used in the agglutinographic reaction chamber and the manufacturing tolerances.

 Reference will now be made to FIG. 4 which shows another embodiment of an agglutinographic reaction chamber, generally indicated at 35, constructed according to the invention. The chamber 35 consists of an upper glass panel 40 and a lower glass panel 41 separated by a distance D-D. The embodiment of FIG. 4 is the same as that of FIGS. 1 to 3 except that the upper and lower panels 40, 41 are longer than the panels 20, 21. This increased distance produces an increased trajectory for the displacement of the liquid and a longer active reaction time. With the longer strip, a wider distribution of agglutinations is possible.

 Reference will then be made to FIG. 5 where an agglutinographic reaction chamber 45 constructed according to another embodiment of the invention is shown. The reaction chamber 45 consists of a single lower panel 41 having three upper panels 47a, 47b, 47c of different lengths. In all other functional aspects, the embodiment of Figure 5 is the same as the embodiment of Figure 1.

The use of the three upper panels of different lengths produces a display of different degrees of agglutination for the same reagent. While the embodiment of Figure 5 is shown with three top panels 47a, 47b, 47c, two top panels or four or more top panels can be used depending on the various conditions.

 Reference will now be made to FIGS. 6A, 6B where an agglutinographic reaction chamber, generally indicated at 55, constructed according to another embodiment of the invention, is shown. In this case, an upper panel 50 has a circular opening 52 at its center. The external dimensions of the upper panel 50 and the lower panel 51 are the same.

 In this embodiment, the reagent is deposited, with a pipette, on the lower panel 51 through the opening 52. This configuration allows a radial pattern of agglutination and presents a lesser opportunity for the reagents to spill. Apart from the location of the receiving region for the reagents, this embodiment has the same functional and structural characteristics and benefits as those described above for FIG. 1. In addition, a radial distribution pattern is created.

Reference will now be made to FIG. 7 where an agglutinographic reaction chamber, generally indicated at 65, constructed according to another embodiment of the invention, is shown. Reaction chamber 65 has a lower panel 61 and an upper panel 60a, both
having the same characteristics and benefits of function and structure as those described above for FIG. 1. However, the capillary chamber 65 also has a second upper panel 60b covering the panel 60a and having the same length as the panel 60a.

In this way, two agglutination layers are visually superimposed for a better observation of the agglutination reactions. Spacers 63 are disposed between panels 60a and 60b and spacers 64 are disposed between panels 60a and 61 to maintain these three panels in spaced relationship. The spacers 63 and 64 can be adjusted to create two capillary chambers of the same or different dimensions.

 We will now refer to FIGS. 8A, 8B where an agglutinographic reaction chamber, generally indicated at 75, is shown, constructed according to another embodiment of the invention. The crap of embodiment of FIGS. 8A, 88 is similar, in its structure, to the embodiment of FIG. 1, identical elements having identical reference numerals. One or more strips of chemicals 73, 74 are placed between panels 20, 21 in the path of the liquid reagents in the capillary chamber 15 to cause a desired additional effect. The desired additional effect can be either chemical or visual. It may be desirable to place a reagent in the agglutinographic reaction chamber and to introduce another reagent at the opening 26 of the capillary chamber 15. This can avoid the need to mix the reagents before introducing them into the reaction chamber. . Several different chemicals can be used to determine if the liquid placed at the edge of the capillary plane agglutinates in the presence of different chemicals.

 Consequently, an agglutinographic reaction chamber is provided which requires neither rocking nor swirling; automatically ends at the end of the reaction; prevents wastage of reagents; requires no reaction monitoring; and is visually stable for a long period of time because there is no noticeable evaporation.

Claims (31)

 1. Agglutinographic reaction chamber for reactive liquid particles characterized in that it comprises, in combination, a first transparent panel (21) having a first surface of a first predetermined area, a second panel (20) having a second surface of a second predetermined area, said second surface being coextensive with and covering at least a portion of said first predetermined surface, said second panel being spaced a predetermined distance from said first panel so that the overlapping portions of the first and second surfaces define a capillary chamber (15) for attracting liquid reagents between the overlapping portions of said first and second surfaces when a reactive liquid is introduced to or around the capillary chamber.  2. Chamber according to claim 1 characterized in that the aforementioned second predetermined surface is smaller than the aforementioned first predetermined surface, the surface not covering thus defined, defining a receiving region (22).  3. Chamber according to claim 1 characterized in that at least one of the first and second surfaces is wettable.  4. Chamber according to claim 1 characterized in that at least one of the first and second panels is a glass panel.  5. Chamber according to claim 1 characterized in that the first and second panels are all gods of substantially flat glass panels.  6. Chamber according to claim 5 characterized in that the first and second panels have the same width and the first panel is longer than the second panel to form a reagent receiving region on the first panel.  7. Chamber according to claim 6 characterized in that the reactive liquid particles are attracted into the aforementioned capillary chamber by introduction of the reagents to the receiving region.  8. Chamber according to claim 1 characterized in that the first and second panels are separated at the predetermined distance mentioned above by a spacing means (30).  9. Chamber according to claim 8 characterized in that the aforementioned spacing means is made of a material chosen from the group consisting of paint, silk screen, ink, film and dust.  10. Chamber according to claim 8 characterized in that the spacing means separates the first panel from the second panel by a predetermined distance in the range of 0.1 n to 500 hum.  11. Chamber according to claim 8 characterized in that the aforementioned spacing means separates the first panel from the second panel by a predetermined distance in the range of 2 n to 20 m.  12. Chamber according to claim 8 characterized in that the aforementioned spacing means separates the first panel of the second panel by a predetermined distance in the range of 3jum to 7 un.  13. Chamber according to claim 8 characterized in that the first aforementioned panel is a glass strip of 50 mm x 50 mm x 1 mm and the aforementioned second panel is a glass strip of 50 mm x 40 mm x 1 mm.  14. Chamber according to claim 1 characterized in that the first and second panels have the same width and the first panel is longer than the second, the length of the second panel being substantially greater than the width of the second panel.  15. Chamber according to claim 1 characterized in that the aforementioned second panel comprises a number of panel members (47a, b, c).  16. A chamber according to claim 15 characterized in that the members forming the panels are glass lamellae.  17. Chamber according to claim 15 characterized in that the aforementioned panels (47a, h, c) are rectangular solids of different lengths.  18. Chamber according to claim Icaractérisée in that the aforementioned second panel has an opening (52) substantially in its center, said opening covering the first panel (51).  19. Chamber according to claim 18 characterized in that the opening of the aforementioned second panel is a circular opening.  20. Chamber according to claim 19 characterized in that the external dimensions of the second panel are the same as the external dimensions of the first panel.  21. Chamber according to claim 18 characterized in that the aforementioned first surface is substantially circular and the aforementioned second surface is substantially annular.  22. Chamber according to claim 1 characterized in that it further comprises a third panel (60b) on the opposite surface of the second panel (60a) relative to the first panel (61), said second panel further comprising a third surface with a third predetermined area, said third panel having a fourth surface with a fourth predetermined area and said fourth surface is coextensive with and covers at least a second portion of said third surface, said third panel being spaced a second predetermined distance from said second panel so that said second portion overlying said third and fourth surfaces defines a second capillary chamber for attracting liquid reagents between said third and fourth surfaces when a liquid reagent is introduced into or around the second capillary chamber.  23. Chamber according to claim 22 characterized in that the aforesaid first and second panels are separated at the aforementioned predetermined distance by a first spacing means.  24. Chamber according to claim 23 characterized in that the aforementioned third panel is spaced from the aforementioned second predetermined distance from the aforementioned second panel by a second spacing means.  25. Chamber according to claim 24 characterized in that the first and second spacing means separate the first and second and third and fourth surfaces substantially of equal distances.  26. Chamber according to claim 24 characterized in that the aforementioned first and second spacing means are made of materials chosen from the group consisting of paint, silk screen, ink, film and dust.  27. Chamber according to claim 22 characterized in that the third and fourth predetermined surfaces smt-snsibiejnent ~ geales.  28. Chamber according to claim 22 characterized in that the second, third and fourth predetermined surfaces are substantially equal.  29. Chamber according to claim 1 characterized in that it further comprises a reaction means between the first panel and the second panel.  30. Chamber according to claim 1 characterized in that the reactive liquid particles are a mixture of a polystyrene latex coated with HCG and an antibody of HCG in solution.  31. Chamber according to claim 30 characterized in that the polystyrene latex has a diameter of 0.3hum.