Filtration system and use thereof
The invention relates to a filtration system for use in a method of determining the presence and/or amount of cells, for example, viable cells, in a liquid sample, and to methods of using and manufacturing such a filtration system. The filtration system includes a cup with an upper portion and a ring portion, where the ring portion is separably coupled to the upper portion.
1. A cell capture system for receiving a fluid sample, the system comprising:
(a) a cup comprising:
(i) an upper portion,
(ii) a ring having a periphery, wherein the upper portion is separably coupled to the ring by a frangible connection comprising a circumferential groove that defines a parting plane between the upper portion and the ring so that the upper portion can be separated from the ring upon manual application by a user of a rotational force sufficient to break the frangible connection, and
(iii) a fluid permeable membrane attached to the periphery to produce a fluidic seal between the membrane and the ring, wherein a portion of the membrane is adapted to retain cells thereon; and
(b) a base comprising a wall configured to receive the ring, the periphery of the ring and the wall of the base further comprising a plurality of protrusions and a corresponding plurality of notches, which when engaged with one another resist rotation of the cup relative to the base when the rotational force is created by the user to break the frangible connection.
2. The system of claim 1 , wherein the protrusions are disposed upon the periphery of the ring and the notches are defined by the wall of the base.
3. The system of claim 1 , wherein the cup further comprises at least two latches each of which is adapted to engage the base.
4. The system of claim 1 , wherein the membrane portion (i) defines a plurality of pores having an average diameter less than about 1 μm so as to permit fluid to traverse the portion of the membrane while retaining cells thereon and (ii) is substantially non-autofluorescent when exposed to light having a wavelength in a range from about 350 nm to about 1000 nm.
5. The system of claim 1 , wherein the membrane portion has a flatness tolerance of up to about 100 μm.
6. The system of claim 1 , wherein the cup is adapted to direct a fluid, when introduced into the upper portion, toward the membrane portion.
7. The system of claim 1 , wherein the ring is integrally formed with the upper portion.
8. The system of claim 1 , wherein the frangible connection further comprises a thin wall at an intersection of the upper portion and the ring.
9. The system of claim 1 , wherein the ring comprises a circumferential registration feature.
10. The system of claim 1 , wherein the membrane is at least one of adhered, bonded, heat welded, and ultrasonically welded to the ring.
11. The system of claim 1 , wherein one of the notches is adapted to receive one of the plurality of protrusions having at least one of a width, a height, a thickness, and a spacing different than each of the other protrusions.
12. The system of claim 1 , wherein the base defines a recess adapted to receive a membrane support.
13. The system of claim 12 , wherein the recess defines a plurality of openings adapted to permit the passage of fluid therethrough.
14. The system of claim 1 , wherein the base further comprises a registration feature.
15. The system of claim 14 , wherein the registration feature comprises a depression defined by a surface of the base.
16. The system of claim 3 , wherein the at least two latches are adapted to resist separation of the cup and the base in a plane perpendicular to the parting plane.
17. A method of harvesting cells if present in a fluid sample, the method comprising:
(a) introducing the fluid sample to the upper portion of the cup of claim 1 ; and
(b) permitting the fluid to pass through the membrane portion.
18. The method of claim 17 further comprising, after applying the fluid, separating the upper portion from the ring.
19. The method of claim 18 , wherein separating the upper portion from the ring comprises applying a force sufficient to decouple the ring from the upper portion.
20. The method of claim 19 , wherein applying the force comprises twisting the cup relative to the base.
21. A method of manufacturing a cell capture system of claim 1 , the method comprising the steps of:
(a) providing a ring having a periphery;
(b) securing a fluid permeable member to a periphery to produce a fluidic seal between the membrane and the ring; and
(c) positioning the ring having the membrane secured thereto within a base configured to receive the ring.
22. The method of claim 21 , wherein, prior to step (b), the ring is separably coupled to the upper portion.
23. The method of claim 21 , wherein the positioning step comprises mating the cup with the base in a predetermined circumferential orientation.
24. The method of claim 21 , further comprising, prior to positioning the cup within the base, placing a porous support in a recess formed in the base.