Abstract
The present invention relates to a method for sampling an egg, the method comprising; a) fluid coupling an interior of the egg to a source of pressure, b) controlling the pressure in the interior of the egg by the source of pressure, c) expelling an amount of fluid, in particular allantoic fluid, from the interior of the egg to the exterior of the egg as a result of the pressure in the interior of the egg, and d) collecting at least a portion of the amount of fluid at the exterior surface of the egg.
Claims
1. A method for sampling an egg, the method comprising; a) fluid coupling an interior of the egg to a source of pressure, b) controlling the pressure in the interior of the egg by the source of pressure, c) expelling an amount of liquid, in particular allantoic liquid, from the interior of the egg to the exterior of the egg as a result of the pressure in the interior of the egg, and d) collecting at least a portion of the amount of liquid at the exterior surface of the egg, and wherein the method further comprises making a sample passage in an egg shell for fluid communication between an interior of the egg and an exterior of the egg; and step d) comprises collecting the portion of the amount of liquid at the sample passage.
2. The method according to claim 1, wherein the method further comprises determining a sampling position at the exterior surface of the egg; and step d) comprises collecting the portion of the amount of fluid at the sampling position.
3. The method according to claim 1, wherein the sample passage has a dimension smaller than 1 mm, in particular smaller than 600 pm.
4. The method according to claim 1, wherein the making a sample passage in an egg shell for fluid communication between an interior of the egg and an exterior of the egg comprises providing a number of passages, wherein the number of passages are preferably arranged in a pattern having a triangular or circular shape, and wherein preferably the number of passages are arranged within a surface of 4 mm.sup.2 up to 100 mm.sup.2.
5. The method according to claim 1, wherein making the sample passage comprises disinfecting the egg shell proximate the sample passage, wherein preferably the disinfecting comprises laser processing the egg shell proximate the sample passage.
6. The method according to claim 1, wherein fluid coupling an interior of the egg to a source of pressure comprises coupling the source of pressure to an air cell of the egg.
7. The method according to claim 1, wherein fluid coupling an interior of the egg to a source of pressure comprises making a flow path through the eggs shell to provide a pressure connection between the source of pressure and the interior of the egg.
8. The method according to claim 1, wherein making a sample passage and/or making a flow path comprises processing an outer egg shell and intermediate layers between the outer egg shell and an interior of the egg with different processing steps.
9. The method according to claim 1, wherein the pressure difference is variable over time and preferably the pressure difference is set at a neutral pressure for a neutral period of time and at an active pressure for an active period of time.
10. The method according to claim 1, wherein the fluid coupling the interior of the egg to a source of pressure comprises engaging a contact area of the egg shell, preferably a contact area at the air cell.
11. The method according to claim 1, the method further comprising sensing at least a portion of the egg to obtain sensor data and making the sample passage depending on the sensor data, in particular determine a position of the sample passage depending on the sensor date, wherein sensing the egg comprises at least one or more of imaging at least a portion of the egg and measuring a position of the egg.
12. The method according to claim 1, and further comprising arranging a fluid intake member at the sample position; and step d) comprises collecting the portion of the amount of fluid with the fluid intake member.
13. The method according to claim 12, wherein the fluid intake member is arranged on the exterior of the egg at least before the end of the incubation and is used during incubation.
14. The method according to claim 12, wherein the fluid intake member comprises an absorb organ and the taking in the portion of the amount of fluid is based on capillary action between the absorb organ and the portion of the amount of fluid.
15. The method according to claim 1, comprising pressurizing the interior of the egg for as long as the sample passage is open for fluid communication between the interior of the egg and the exterior of the egg.
16. The method according to claim 1, comprising monitoring the amount of expelled sample fluid to obtain sample fluid amount data and comparing the fluid amount data with a defined minimum amount data and depending on the step of comparing, repeating, intensifying or maintaining at least step c), or closing the sample passage.
17. The method according to claim 1, comprising closing the sample passage to stop fluid communication between the interior of the egg and the exterior of the egg.
18. The method according to claim 17, wherein closing the sample passage comprises contacting the sample passage with a closure element and the method comprises depressurizing the interior of the egg after contacting the sample passage with the closure element in order to increase a closing contact between the closure element and the sample passage.
19. The method according to claim 17, wherein closing the sample passage comprises manipulating an egg in order to force an intermediate layer between the outer egg shell and the interior of the egg, towards the sample passage.
20. The method according to claim 1, comprising maintaining the egg in a predetermined position during a settling time before expelling the amount of fluid from the interior of the egg to the exterior of the egg.
Description
DESCRIPTION OF THE DRAWINGS
(1) The invention will be further elucidated referring to the schematic drawings wherein shown in:
(2) FIG. 1A-G in side view a number of embodiments of the method for sampling an egg according to the invention;
(3) FIG. 2A, 2B, 3A, 3B show a detail of an egg and examples of fluid intake members that collect an amount of fluid;
(4) FIG. 4A in side view processing steps for making a sample passage in the egg shell;
(5) FIG. 4B shows a detail of an egg after the processing step of steps of FIG. 4A;
(6) FIG. 5A, 5B show embodiments of a step of sensing an egg;
(7) FIG. 6A-D show different examples of pressure versus time graphs of pressure in the interior of the egg;
(8) FIGS. 7A and 7B show an embodiment of a process of closing the sample passage;
(9) FIG. 8 shows another embodiment of a process of closing the sample passage;
(10) FIGS. 9A and 9B show a further embodiment of a process of closing the sample passage; and
(11) FIGS. 10A and 10B show an even further embodiment of a process of closing the sample passage 10.
DETAILED DESCRIPTION OF EMBODIMENTS
(12) FIG. 1A-E show embodiments of the method for sampling an egg according to the invention.
(13) FIG. 1A shows a cup 1 that is designed to fluid couple with the shell 2 of an egg 3. The cup 1 is fluid coupled with a source of pressure (not shown here). As a result, an interior 4 of the egg 3 is fluid coupled to the source of pressure through the cup 1 and because of the porosity of egg shell 2. An egg shell 2 has pores (not shown) for gas exchange between the interior 4 of an egg 3 and the exterior 6. These pores have a diameter between 1 to 10 microns.
(14) As soon as the interior 4 of the egg 3 is fluid coupled to the source of pressure, the pressure in the interior 4 of the egg 3 can be controlled by the source of pressure. The amount of fluid 7 is expelled from the interior 4 of the egg 3 to the exterior 6 of the egg 3 as a result of overpressure applied to the cup 1. In other words, the cup 1 is pushed on the egg 3 and an overpressure is applied to the suction cup 1.
(15) A pressure difference between the interior 4 of an egg 3 and the exterior 6 results in transport of fluid through the egg shell 2. Here, the pressure in the interior 4 exceeds the atmospheric pressure at the exterior 6 and as a result an amount of fluid 7 is expelled from the interior 4 of the egg 3 to the exterior 6 of the egg 3. In this case, the expelled amount of fluid 7 is allantoic fluid.
(16) Downstream in the process of the method, a portion of the amount of fluid 7 is collected at the exterior surface 2 of the egg 3, as is best shown in FIG. 2, 3.
(17) Once the egg is sampled, any desired analyses can be executed with the collected portion of the amount of fluid 7, like determining the gender of the embryo situated in the egg 3.
(18) Method according to a preceding claim, wherein controlling the pressure in the interior of the egg by the source of pressure comprises applying a pressure difference between the interior and the exterior of the egg
(19) Method according to claim #, wherein the fluid coupling the interior of the egg to a source of pressure comprises engaging a contact area of the egg shell, preferably a contact area at the air cell
(20) FIG. 1B-G show different embodiments of the method for sampling an egg according to the invention. In general, only differences compared with FIG. 1A are described.
(21) FIG. 1B shows the cup 1 fluid coupled with the shell 2 of an egg 3. In the egg shell a flow path 9 is made by machine action to the egg shell 2. The cup 1 is positioned over the flow path 9 in order to fluid couple the interior 4 of the egg 3 to the source of pressure. The flow path 9 works in parallel to the pores that are present in the egg shell 2. The flow path 9 is made at the air cell 5 of the egg 3. The air cell 5 is a convenient position to breach the protection that the egg shell 2 offers to an embryo in the interior 4 of the egg 3. A reason therefore is a membrane 8 that separates the air cell 5 form the remainder of the interior 4 of the egg 3. The membrane 8 is flexible so therefore, the interior 4 of the egg can be pressurized through the air cell 5.
(22) FIG. 1C shows the cup 1 fluid coupled with the shell 2 of an egg 3. In the egg shell a sample passage 10 is made by machine action to the egg shell 2. The sample passage 10 facilitates fluid communication between the interior 4 of the egg 3 and an exterior 6 of the egg 3. The sample passage 10 works in parallel to pores that are present in the egg shell 2. The sample passage 10 may a diameter smaller than 1 mm, in particular smaller than 600 ?m which is a big passage compared with pores that have a diameter of 1 to 10 microns. The portion of the amount of fluid 7 is collected at the sample passage 10. The sample passage 10 is provided near the air cell 5 but however past the membrane 8 as seen from the air cell 5. The membrane 8 separates the air cell 5 and the sample passage 10. The sample passage 10 is therefore positioned at the locality where allantoic fluid accumulates.
(23) The sample passage 10 may have a cylindrical shape, however a conical shape that tapers toward the exterior 6 of the egg 3 is conceivable as well.
(24) Where one sample passage is shown, it will be conceivable that a number of passages can be provided. These number of passages can be arranged in a pattern. In case of three or more passages, the pattern may have a triangular shape or circular shape. The number of passages are arranged within a surface of 4 mm.sup.2 up to 100 mm.sup.2 to facilitate collecting of a portion of the amount of fluid 7.
(25) FIG. 1D shows the cup 1 fluid coupled with the shell 2 of an egg 3. In the egg shell 2 a sample passage 10 is made as well as the flow path 9.
(26) FIG. 1E shows the cup 1 fluid coupled with the shell 2 of an egg 3. The cup 1 is positioned near the air cell 5 but however past the membrane 8. The cup 1 is therefore positioned at the locality where allantoic fluid accumulates. In this case, the expelling the amount of fluid 7 from the interior 4 of the egg 3 to the exterior 6 of the egg 3 as a result of underpressure applied to the cup 1. In other words, the cup 1 operates as a suction cup 1. The amount of fluid 7 is expelled within the inner of the cup 1. Although not shown, it will be clear that in this case the sample passage 10 can be made in the egg shell 2 as well if desired.
(27) FIG. 1F is similar to FIG. 1A or 1E. In this case, the expelling the amount of fluid 7 from the interior 4 of the egg 3 to the exterior 6 of the egg 3 is caused by a temperature increase of the egg 3 that is shown with a temper. The temperature of the interior 4 of the egg 3 can be increased by any suitable means like based on microwave action. In addition, the cup 1 can be applied to the egg 3 as well to close off the pores in the egg shell at the air cell and/or to facilitate the expelling the amount of fluid 7 from the interior 4 of the egg 3 to the exterior 6 of the egg 3.
(28) FIG. 1G is similar to FIG. 1C. The egg 3 is rotated along its longitudinal axis 12. In this case, the egg 3 is rotated over about 180? however any angular position will do as long as gravity helps to expel allantoic fluid through the sample passage 10. As a result the sample passage 10 faces downwards. This facilitates fluid communication between the interior 4 of the egg 3 and an exterior 6 of the egg 3. The reason therefor is that the egg content helps to expel allantoic fluid through the sample passage 10 because of gravity. Thus, firstly the allantoic fluid accumulates during a settling time wherein the egg is maintained in a predetermined position shown fin FIG. 1A. Then, the egg is rotated as shown and the amount of fluid 7 is expelled from the interior 4 of the egg 3 to the exterior 6 of the egg 3.
(29) FIG. 2A, 2B, 3A, 3B show a detail of an egg 3 and examples of fluid intake members 13, 14 that collect a portion 15 of the amount of fluid 7.
(30) In FIGS. 2A and 2B a fluid intake member 13 is in the form of a tissue paper that functions as an absorbing organ. The fluid intake member 13 is attached to the egg shell 2 of the egg 3. The fluid intake member 13 can be arranged on the exterior of the egg 3 at least before the end of the incubation and be used during incubation. The fluid intake member 13 can be arranged on the exterior of the egg 3 during sampling of the egg 3. The fluid intake member 13 can be arranged on the exterior of the egg 3 at the start of the incubation and be used during incubation. As an option, the fluid intake member 13 can be removed before hatching. The fluid intake member 13 is attached to the egg 3 past the membrane 8 as seen from the air cell 5. In this case, the fluid intake member 13 is attached to the egg 3 near the air cell 5 past the membrane 8. The shown position of the fluid intake member 13 near the air cell 5 is also referred to as the sample position. The fluid intake member 13 is therefore positioned at the locality where allantoic fluid 11 accumulates in the egg 3. A portion or all of the amount of fluid 7 is collected with the fluid intake member 13 by absorption. FIG. 2B differs with FIG. 2A in that a sample passage 10 is provided in the egg shell 2. The fluid intake member 13 cover the sample passage 10. In this case, the fluid intake member 13 covers the entire sample passage 10.
(31) In FIGS. 3A and 3B a fluid intake member 14 is in the form of a capillary tube. The fluid intake member 14 is attached to the egg shell 2 of the egg 3. The fluid intake member 14 approaches to the egg 3 near the air cell 5 but however past the membrane 8. The shown position of the fluid intake member 14 near the air cell 5 is also referred to as the sample position. The fluid intake member 14 is therefore positioned at the locality where allantoic fluid 11 accumulates in the egg 3. FIG. 3B differs with FIG. 3A in that a sample passage 10 is provided in the egg shell 2. A portion 15 of the amount of fluid 7 is collected with the fluid intake member 14 by capillary action.
(32) FIG. 4A shows in side view processing steps for making a sample passage 10 in the egg shell 2. Making the sample passage 10 comprises laser processing in a laser processing unit 16, and/or processing in a machining unit 17 like a puncturing, cutting, milling or drilling unit. The egg 3 is transported in a process flow direction 18 along the laser processing unit 16, and/or processing in a machining unit 17. It will be clear that the processing steps may also apply to the making of the flow path 9. As an option, the making the sample passage 10 may comprise disinfecting the egg shell proximate the sample passage 10. The disinfecting may comprise laser processing the egg shell 2 proximate the sample passage 10 using the laser processing unit 16.
(33) When making a sample passage 10 and/or making a flow path 9, it is possible that an outer egg shell 2 and intermediate layers need to be crossed. It is conceivable that the outer egg shell 2 and intermediate layers, like for example and if required the membrane 8, are processed with different processing steps.
(34) FIG. 4B shows a detail of an egg 3 after the processing steps for making a sample passage 10 in the egg shell 2. Two possible configurations of tapered sample passages 29. 30 are shown in cross sectional side view. The sample passage 29 tapers out towards the interior 4 of the egg 3. This minimize the area of the sample passage 29 at the outer surface of the egg shell 2 which prevents ingress of pollution. In addition, the sample passage 29 is not easily obstructed by the shell membrane (not shown) that may shift a little with respect to the shell 2. The configuration of passage 29 is in particular enabled by the laser processing unit 16.
(35) The sample passage 30 tapers inward towards the interior 4 of the egg 3. This minimize the area of the sample passage 29 at the inner surface of the egg shell 2 which reduces the risk to damage the content of the egg 3 like important blood vessels.
(36) FIG. 5A, 5B show embodiments of a step of sensing the egg 3. A sensor unit 19 is provided to sense the egg 3. The sensor unit 19 is configured to survey an egg and/or for monitoring the amount of expelled sample fluid. The sensor unit 19 may comprise any suitable sensing means like an image capturing device such as a camera. The sensor unit 19 is operationally coupled with a source of pressure 20, in this case a controllable source of pressure 20.
(37) In an egg surveying mode, the following steps are executed; determining a sampling position at the exterior surface 2 of the egg 3; and collecting the portion of the amount of fluid at the sampling position. The sensor unit 19 maps the egg 3 for items like allantoic fluid, the embryo, blood vessels etc. The sampling position is then based on egg sensor data and is normally proximate accumulated allantoic fluid in the egg 3.
(38) In a sample passage making mode, the following steps are executed; sensing at least a portion of the egg 3 to obtain sensor data and making the sample passage 10 depending on the sensor data, in particular determine a position of the sample passage depending on the sensor data. Here, sensing the egg 3 may comprise imaging at least a portion of the egg 3 and measuring a position of the egg 3.
(39) In a sample monitoring mode, the following steps are executed; monitoring the amount of expelled sample fluid 7 to obtain sample fluid amount data and comparing the fluid amount data with a defined minimum amount data and depending on the step of comparing, repeating or maintaining expelling allantoic fluid from the interior 4 of the egg 3 to the exterior 6 of the egg 3 as a result of the pressure in the interior 4 of the egg 4. If enough amount of fluid is expelled, the sample passage 10 can be closed to stop fluid communication through the sample passage 10. In the sample monitoring mode, the sensor unit 19 can be orientated towards the capillary tube 14 in order to directly monitor the portion 15 of the amount of fluid 7 in the capillary tube 14.
(40) FIG. 6A-D show different examples of pressure versus time graphs of pressure in the interior of the egg 3. In all graphs, the atmospheric pressure is referred to with reference number 21. All FIG. 6A-D show that the pressure difference is variable over time.
(41) FIG. 6A shows two subsequent periods 20 of over pressure in the interior 4 of the egg 3.
(42) FIG. 6B shows two subsequent periods 22, 23 of over pressure in the interior 4 of the egg 3. The pressure is increased in the second period of time 23 compared with the first period of time 22. FIG. 6B is an example of pressurizing the interior 4 of the egg 3 at an overpressure for as long as the sample passage 10 is open for fluid communication between the interior 4 of the egg 3 and the exterior 6 of the egg 3. Once the sample passage 10 is closed, the pressure can be released to the atmospheric pressure.
(43) FIG. 6C shows consecutive an active period 24 where over pressure is applied, a neutral period 25 wherein a neutral pressure is applied, and another active period 26 where under pressure is applied to the interior of the egg. The neutral pressure is normally atmospheric pressure that prevails at the exterior 6 of the egg 3.
(44) FIG. 6D shows a period 27 of under pressure in the cup 1 that can be applied to the sample passage 10.
(45) FIGS. 7A and 7B show an embodiment of a process of closing the sample passage 10. The sample passage 10 is closed after expelling the amount of fluid 7 from the interior 4 of the egg 3 to the exterior 6 of the egg 3 as a result of pressure difference. The closing the sample passage 10 stops fluid communication between the interior 4 of the egg 3 and the exterior 6 of the egg 3. The sample passage 10 is closed to prevent ingress of pollution through the sample passage 10. Closing the sample passage 10 comprises contacting the sample passage 10 with a closure element 28. The closure element 28 is here a micro bead 28. A number of micro beads 28 are disposed on the amount of fluid 7. When the amount of fluid 7 is withdrawn back into the interior 4 of the egg 3, the beads 10 are taken with the amount of fluid 7 towards the sample passage 10. At least one micro bead 28 will close off the sample passage 10. The micro bead 10 is configured to close off a sample passage 10. The amount of fluid 7 can be withdrawn back into the interior 4 of the egg 3 by applying a suitable pressure difference. For example, an under pressure can be applied to the interior 4 of the egg 3 or an over pressure can be applied at the sample passage 10. The pressure difference between the interior 4 and the exterior 6 of the egg 3 increases closing contact between the micro bead 28 and the sample passage 10.
(46) FIG. 8 shows another embodiment of a process of closing the sample passage 10. Closing the sample passage 10 comprises contacting the sample passage 10 with a closure element 28. The closure element 31 is here an adhesive member, in this case a sticker 31. The sticker 31 covers the sample passage 10 and is in sealing contact with the egg shell 2 around the sample passage. As an option, the fluid intake members 13 is integrated with the sticker 31. The sticker 31 may be transparent, or at least have a transparent portion to enable line of sight to the fluid intake member 13.
(47) FIGS. 9A and 9B show a further embodiment of a process of closing the sample passage 10. The closure element 34 is here a valve member 34. The valve member 34 is moveable between a sample passage open position shown in FIG. 9A and a sample passage closing position in shown in FIG. 9B. The valve member 24 is preferably a normally closed type of valve member. The valve member 34 is part of a valve device 32. The valve device 32 has a valve support 33. The valve support 33 couples with the egg shell 2. The valve support 33 maintains the valve member 34 at the sample passage 10. The valve member 34 is moveably coupled with the valve support 33. Here, the valve member 34 is moveably coupled with the valve support 33 through a living hinge construction. The valve member 34 is like the micro bead 28, operated by pressure difference. The pressure difference between the interior 4 and the exterior 6 of the egg 3 increases closing contact between the valve member 34 and the sample passage 10. As an option, the fluid intake members 13 is integrated with the valve device 32.
(48) FIGS. 10A and 10B show an even further embodiment of a process of closing the sample passage 10. Here, the closing of the sample passage 10 comprises manipulating an egg in order to force an intermediate layer 35 between the outer egg shell 2 and the interior of the egg, towards the sample passage 10. Manipulating of the egg 3 may include moving, shaking, twisting etc., to make the egg contents move with respect to the egg shell 2. The intermediate layer 35 is the egg shell membrane that closes off the sample passage 10 as shown in FIG. 10 B.
