The invention relates to a device with which conjunctival impressions may be taken.
The conjunctival impression technique was developed at the end of the 1970s and is now an accepted technique allowing conjunctival epithelial cells to be collected from the conjunctival mucosa which is an immunologically reactive and complex tissue, in a quasi-painless and non-invasive way, for purposes of analyses and diagnose. The most superficial cells of the conjunctival epithelium, i.e. the cells undergoing desquamation, are collected by this technique. Conjunctival epithelium renews itself from the basal layers, and the most superficial cells which have reached their final differentiation are regularly discharged into the lachrymal fluid. It is this surface layer comprising the terminally differentiated conjunctival cells, which may be sampled by the conjunctival impression for subsequent analysis.
There are three types of cell populations which may be found on conjunctival impressions:
With conjunctival impressions, various cell populations, their size, number, density and pathological modifications may be analyzed and important information may be provided on the status of the eye surface. They also provide information on the conjunctival attack, notably in the field of the dry eye, iatrogenic pathologies induced by collyria, pathologies of the limbus and chronic inflammation of the eye surface.
In order to take a conjunctival impression, the most current present method consists in using membrane filters commonly used in biochemical industry, the most often in cellulose ester (a mixture of cellulose ester and cellulose nitrate) or in polyethersulfone, for example, and which are applied on the conjunctiva, generally without any local anesthesia. The sampling is totally bloodless, painless and non-invasive contrary to other more aggressive systems such as brushing or biopsy. During the sampling, the filter is generally applied on the upper bulbar conjunctiva of the eye, protected by the eyelid and removed after a contact of 3-5 seconds. The desquamating conjunctival cells then adhere to the membrane surface and form a thin and homogenous cell layer. The filters are then treated in order to recover the cells to be analyzed.
Within the scope of scientific evaluations or when the examinations will be repeated, the impression should always be taken at the same location because there are disparities in the distribution of the sampled cells on the bulbar conjunctiva. This is neither obvious nor easy with the present sampling methods which remain relatively manual even today, in spite of various attempts to improve the procedure (different materials, porosities, holding devices . . . etc.). They are relatively cumbersome and time-consuming for a clinician or a pathologist within the scope of routine daily use.
An object of the invention is to provide a device for taking an eye impression which is simple to implement while supplying reliable and reproducible sampled impressions.
For this purpose, provision is made according to the invention for, an eye sampling device comprising supporting means comprising a sampling surface, the supporting means being elastically deformable between a rest position and a sampling position and conformed so that, upon being used for sampling, the sampling surface applied in the sampling position in a coinciding way onto a sampling area (2;402;502), is detached therefrom according to a peeling movement when passing from the sampling position to a rest position.
Thus, by the use of elastically deformable supporting means bearing the sampling surface and conformed in order to allow the latter to be detached from the eye sampling area according to a peeling movement, eye impressions which are of optimum quality, may be obtained reliably and reproducibly. Further, this allows a sampling device to be produced, which is simple to use.
Advantageously, but optionally, the device according to the invention has at least one of the following features:
Other features and advantages of the invention will emerge from the description hereinafter of a preferred embodiment and also of variants. In the appended drawings:
a and 1b are schematic views of the sampling kinematics achieved by a device according to the invention;
a and 2b are schematic views of alternative embodiments of the deformable portion of the device according to the invention;
a-3c are schematic views of alternative embodiments of the sampling surface of the device according to the invention;
a and 7b are three-dimensional schematic views of alternative shapes of impressions taken by a device according to the invention; and,
As a liminary remark, the sampling area 2 of an eye impression is located on the conjunctiva, an area located at the periphery of the cornea, which extends from the limbus which delimits the junction between the cornea (nominal diameter of the cornea: about 12 mm for an adult) and the sclera, and extends outwardly to an area externally delimited by a circle with a diameter of about 22 mm and centered on the cornea. For example, as illustrated in
The area 2 for taking an eye impression is moreover located on the eyeball which may be assimilated to a sphere with a diameter of 24 mm+/−1 mm, except at the cornea. The eye's anatomy is recalled and illustrated in
With reference to
This kinematics is important in order to ensure reliability and reproducibility as well as quality of the eye impressions taken by means of the device 10 according to the invention in order to maximize the quality of the sampled cells while maintaining their integrity.
As a matter of principle, the device 10 according to the invention includes a piston forming component 30 having an end 22 on which supporting means 12 are attached. These supporting means 12 are elastically deformable between a rest position illustrated in
This sampling movement is obtained by presenting the sampling surface 11 in a non-parallel or non-coinciding way towards the sampling area 2 of the eye surface. Thus, the sampling surface 11, defined by a planar geometry here, is presented so as to face the sampling area 2 of the eye surface according to a given orientation, so that, at the time of contact between both surfaces, there is no coincidence of these surfaces. Indeed, contact between the sampling surface 11 and the sampling area 2 of the eye 1 is initiated by the contact of the low point (or low line) 121 on said sampling area and proceed by moving this contact point (or line) along the sampling surface 11, by sweeping through the latter, right up to the high point (or high line) 122 which lastly comes into contact with the sampling area 2, and this under the effect of a movement along the longitudinal axis of the device 10 according to the invention as illustrated by the arrow of
The sampling support, once it is placed on the eye 1, is then detached from the eye 1 in the least traumatic way, by exerting a peeling movement. Indeed, with a stress mode of the peeling type, concentrated stresses may exist at the location of the detachment which, here, is a point on the schematic view of
The supporting means 12 are made in a block of elastically deformable material such as a foam or a suitable elastomer (elastic polymer): polyurethane, polyethylene, silicone foam, a thermoplastic elastomer such as Styrene-ethylene/butylene-styrene (SEBS), Styrene-ethylene/propylene-styrene (SEPS) ethylene-propylene copolymer/polypropylene (EPDM-PP), polyether block amides (PEBAX), polyurethane or further a vulcanizable elastomers such as silicone rubber, latex, polybutadiene, a fluorinated elastomer, polychloroprene, polyisoprene . . . etc.
Two alternative embodiments of the supporting means are illustrated in
Likewise, with reference to
For these different geometries, the sampling surface 111,211,311 always has a low point and at least one high point in order to be able to allow the sampling movement according to the kinematics described earlier, with reference to
Upon removing or detaching the sampling surface, the progressive movement of this same dynamic contact line is the inverse of the movement performed during the contacting.
Thus, the detachment movement is analogous to a peeling movement with which the cells of the sampling area 2 may be gradually detached from the eye surface, which have then adhered onto the sampling support of the sampling surface. This adhesion on the sampling support is achieved according to different well-known principles: by hydrophilicity, by standard surface tension, by adhesion of the cell mucus, or anionic electrostatic charges of the cells.
With reference to
This sampling surface 11 includes the sampling support intended to collect an eye impression upon sampling on a sampling area 2 of the eye 1. The sampling support includes at the periphery, around the sampling surface 11, a flexible edge 13, 14 (added by insert-molding, adhesive bonding, welding . . . etc.), this edge having a protrusion directed radially towards the supporting means 12, which will fit onto a notch 15, 16 located on the perimeter of the supporting means 12. Once it is in place, the edge 13, 14 extends and protrudes from the supporting means 12.
In the case illustrated in
The piston 20 is slidably mounted in a body 30 of the device 10 according to the invention and this against an elastic return means 21, here as a compression spring. The body 30 includes a first radially inner wall including an end 32 intended to come into contact with the eye 1 during sampling. Additionally, the radially inner wall of the body 30 includes a radially outer shoulder 34, optionally with a frusto-conical shape, the role of which will be described subsequently with reference to
Given that the conjunctiva is a very mobile tissue relatively to the underlying sclera S, that the eye 1 is capable of reflex eye movements upon sampling (with the risk of damaging the sample by a shearing effect between the sampling support placed on the conjunctiva and the conjunctiva), that the conjunctiva is attached at the limbus L and since there is a change in the radius of curvature between the cornea C and the sclera S, the end 32 of the device 10 according to the invention includes a semi-circular supporting area intended to come into contact, by fitting its contour, with the limbus L of the eye 1, thereby ensuring proper positioning on the one hand and properly maintaining the relative position between the eye and the device according to the invention during the sampling, on the other hand.
The body 30 further includes a second radially outer wall comprising an end 31 intended to come into contact with the eye 1 during sampling. Additionally, the radially outer wall of the body 30 includes a radially inner shoulder 33, the role of which will be described subsequently with reference to
Given that the edges of the eyelids are often contaminated by bacterial strains different from those present at the eye surface, there is a risk of contamination of the taken sample specifically dedicated to the analysis of this surface. Consequently, before putting the sampling support in contact with the sampling area of the eye, the surface including the sampling area of the edge of the eyelids has to be isolated. This is the role of the end 31 which is conformed in order to face the eyelid, by being accommodated below the eyelid, while maintaining the latter open and preventing any crossed contamination between the sample and the eyelid. For this, the end 31 has a foot-shaped section, the sole of which has a section of a concave shape mating that of the eye surface upon which the end 31 is intended to bear during sampling, the eyelid will then come into contact and be retained by the top of said foot.
On the other hand, it should be noted that the eye surface is covered with a lachrymal film consisting of tears and of a thin layer of lipids secreted by the Meibomian glands located at the lid margin of the lower and upper eyelids. This thin lipidic and hydrophobic layer will limit or prevent adhesion of the cells on the highly hydrophilic surface of the sampling support. Consequently, before having the sampling support come into contact with the sampling area of the eye, the tear film has to be broken. This is the additional role of the ends 31 and 32 of the body 30. For this, these ends 31 and 32 are made in or covered with a semi-rigid or elastomeric (so as not to injure the eye surface and to achieve intimate contact with the latter) material and which is both absorbent and lipophilic so as to absorb the lipid portion of the tear film at the sampling area of the sampling surface thereby delimited by these ends 31 and 32.
With reference to
The device 10 according to the invention is placed on the eye 1 so that the end 32 of the body 30 will contact the limbus L and that the end 31 of the body 31 is inserted under the eyelids. Next, pressure is exerted on the end 23 of piston 20 in order to accomplish the sampling. This pressure is exerted along the longitudinal axis X-X and oriented towards the eye (see the arrow of
Now the sampling support 11 remains to be recovered in order to be able to carry out the intended analyses. Recovery of the sampling support 11 is performed after having placed at least the portion including the sampling support 11 of the device according to the invention inside a collecting means, such as for example a collecting tube.
The piston 20 is then pulled backwards according to the arrow of
Alternatively, the detachment of the sampling support 11 is carried out according to a gradual movement defined by the initial position of the shoulders 33 and 34 along the travel covered by the piston 20 when it is pulled backwards in the body 30: the edge 14 will first bear upon the shoulder 33 and its protrusion disengages from the notch 16, and then the edge 14 will first bear upon the shoulder 34 and its protrusion disengages from the notch 15, thereby releasing the sampling support 11. For this, the shoulders 33 and 34 are not located at a same height along the axis of the device and the edges 14 and 13 themselves are not at the same height, which is the case in the example of
The sampling support 11 falls into the collecting means without any other handling from behalf of the user and with very limited contact with open air. This thereby reduces the risks of accidental contamination (desirable for analyzing the germs present at the eye surface and detecting infestations of microorganisms, for example) and of damage before utilization.
Indeed, cytology for example requires the cells to be very well attached onto the sampling support 11 so as to be marked with fluorescent markers and allow observation by conventional microscopy (with transparization of the support) or confocal microscopy.
Cytometry as for it, requires the cells to adhere as much as possible on the support, while allowing to be easily detached when desirable into a collecting solution with order to proceed to cell counting and sorting.
Finally, sample analysis by means of polymerase chain reaction (PCR), assumes destruction of the sampling support into the products used for cell lysis allowing DNA extraction.
By using the device according to the invention, eye impressions may be optimized to the various kinds of analysis techniques by using different supports adapted to the analysis procedure.
The sampling support 11 is here an insert-molded membrane filter type with a flexible plastic edge (which has the advantage of being a versatile solution, allowing the use of several types of membranes). Alternatively, it is possible to mould a shaped one-piece part (membrane+edge) in a plastic material ad hoc, and if necessary including a treated surface, optimized for the sampling (roughness, plasma treatment in order to increase hydrophilicity, . . . etc.) in order to produce the sampling support.
Considering the foregoing, the device 10 according to the invention therefore provides control of the two following parameters when placing and removing the sampling support, for making the samplings more reliable:
Further, with the device 10 according to the invention, as described earlier, the problem of the tear film may be solved, and the sampling surface may be defined and localized always at the same location and the eye may be immobilized, as well as recovery may be optimum without damaging the taken eye impression.
As alternative embodiments, the device according to the invention, which has just been described according to one embodiment, where it has an axisymmetrical ring shape, may appear in any geometrical (sector, round, oval, ovoid . . . etc.) shape able to be inscribed between the diameter at the limbus and the diameter of about 22 mm.
A first alternative embodiment illustrated in
A second alternative embodiment illustrated in
In both of these alternative embodiments, as well as in others where the device according to the invention does not have an axisymmetrical ring shape, the shoulders 33 and 34 form a single and same shoulder inside the body of the device, as well as the notches 13 and 14 which are only a single notch at the periphery of the supporting means 12.
The device according to the invention described earlier therefore operates by having the conjunctiva come into contact with a material allowing controlled adhesion of the cells. Most particularly, it ensures that:
Additionally with it, it is possible to:
The material allowing the sampling and forming the sampling support 11 (foam, woven or non-woven fabric filter, hydrogel, polymer whether treated or not . . . etc.) is compatible with the present methods of analysis, i.e:
Other exemplary uses of the taken sample may relate to recovery of DNA (within the scope of forensic medicine for example) or to the analysis of germs present at the eye surface (viruses and bacteria).
It is also possible to search for atmospheric pollutants or even radioactive particles in the sample and it would thus be simpler to measure contamination than by means of a blood sample. When the eye is open, it is continually in contact with its environment and exposed to airborne substances or to substances present in the atmosphere, and in some way, by collecting these substances, it is acting as a sensor. Collection of substances collected by the eye with this method may have advantages as compared with other techniques used such as skin biopsies or blood samples. More generally, it is/will be possible to collect and also analyze the composition of tears and to detect the content of chemical mediators, hormones (insulin, . . . ), DNA, blood proteins, ions, enzymes, in normal and/or pathological situations.
In view of the teaching contained herein, one skilled in the art can identify various other configurations to the invention without departing from the scope thereof.
Number | Date | Country | Kind |
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09305209.0 | Mar 2009 | EP | regional |
Filing Document | Filing Date | Country | Kind | 371c Date |
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PCT/EP2010/052832 | 3/5/2010 | WO | 00 | 9/2/2011 |