Multilevel disposable cartridge for biological specimens
A multilevel, disposable cartridge may have a plurality of fluid wells or reservoirs holding a plurality of reagents, and a sample viewing area to view a biological sample. The reagents may be applied sequentially to the sample by opening a specific valve formed in an elastomeric, flexible layer under the fluid well. The fluid channels may be formed in a first rigid plastic layer to conduct the reagent to the sample. Pneumatic channels for applying suction or pressure to the valves may be formed in a second rigid, plastic layer.
1. A disposable for analyzing biological specimens, comprising:
a first rigid layer ( 20 ), having an analyzing area ( 24 ), a plurality of fluid reservoirs ( 22 ) and a plurality of fluidic channels formed therein wherein the channels provide fluid communication to the fluid reservoirs;
a flexible layer ( 30 ) having a plurality of fluidic control points ( 33 ) in fluid communication with at least one of the fluid reservoirs ( 22 ); a second rigid layer ( 40 ) with a plurality of pneumatic channels formed therein, wherein the channels provide pneumatic communication to the at least one fluid control point; wherein each fluidic control point is pneumatically controllable to cause flow from only one of the plurality of fluid reservoirs ( 22 ) into or out of the fluidic channels, wherein each fluidic control point ( 33 ) is pneumatically controllable to cause flow from only one of the plurality of fluid reservoirs ( 22 ) into or out of the fluidic channels ( 125 ) characterized in that the second rigid layer ( 40 ) further comprises an attachment mechanism ( 45 ) for holding a support ( 60 ), wherein the support ( 60 ), the rigid layers ( 40 , 20 ) and the flexible layer ( 30 ) define a transparent sample analyzing volume ( 500 ) for imaging the biological specimens in the sample analyzing volume, and wherein the first rigid layer ( 20 ) further comprises a straight trench ( 220 ) adjacent to at least one side of the sample analyzing volume ( 500 ), which accepts and distributes fluid flowing from at least one fluidic reservoir ( 22 ) over a whole side of the analyzing volume ( 500 ) and wherein the fluidic control point ( 33 ) comprises a thinned portion ( 133 ) of the flexible layer ( 30 ), and wherein the straight trench ( 220 ) comprises two parallel straight trenches ( 220 ) disposed on two sides of the sample analyzing volume ( 500 ).
2. The disposable of claim 1 , wherein the second rigid layer ( 40 ) further comprises an attachment mechanism ( 45 ) for holding a support ( 60 ), wherein the support ( 60 ), the rigid layers ( 46 , 20 ) and the flexible membrane ( 30 ) define a sample analyzing volume ( 500 ).
3. The disposable of claim 1 , wherein the fluidic control points are pneumatically controlled by suction applied from a pneumatic channel of the second rigid layer against a backside of the flexible layer, wherein the flexible layer is withdrawn from a first stop against a second stop allowing fluid to flow in and out of the-sample analyzing volume ( 500 ).
4. The disposable of claim 2 , wherein the fluidic control points are pneumatically controlled by suction applied from a pneumatic channel of the second rigid layer against a backside of the flexible layer, wherein the flexible layer rests against a first stop preventing fluid to flow in and out of the sample analyzing volume ( 500 ).
5. The disposable of claim 1 , wherein the fluidic control points comprise a thinned portion ( 133 ) of the flexible layer ( 30 ) which is deflected against a first stop ( 120 ) and a second stop ( 43 ).
6. The disposable of claim 5 , wherein the fluid control point comprises a raised feature ( 120 ) in the first rigid layer ( 20 ) which protrudes into the fluidic channel, and defines first stop ( 120 ) for the thinned portion ( 133 ) of the flexible layer ( 30 ) and an opening in a pneumatic channel of the second rigid layer ( 40 ) as second stop ( 43 ) for the thinned portion ( 133 ) of the flexible layer ( 30 ) wherein the thinned portion ( 133 ) opens and closes the fluidic channel in the first rigid layer ( 20 ).
7. The disposable of claim 5 , wherein the second rigid layer ( 40 ) comprises a pressure point ( 43 ) which protrudes into a cavity of the flexible layer ( 30 ) as second stop for the thinned portion ( 133 ) of the flexible layer ( 30 ).
8. The disposable of claim 7 , wherein the pressure point ( 43 ) protrudes into the cavity, thereby registering the flexible membrane ( 30 ) with respect to the rigid layer ( 40 ), at a location adjacent to a button formed on the thinned portion ( 133 ) of the flexible layer ( 30 ).
9. The disposable of claim 1 , wherein the flexible layer ( 30 ) further comprises a pumping mechanism ( 200 ) actuated by a pneumatic force.
10. The disposable of claim 9 , wherein the pumping mechanism ( 200 ) comprises a movable part ( 210 ) in the flexible layer ( 30 ) configured to alter the volume of a fluidic channel, and wherein said movable part ( 210 ) is in fluidic communication between a first ( 100 ) and a second fluidic control point ( 300 ).
11. The disposable of claim 2 , wherein the second rigid layer comprises a first pneumatic channel providing suction to a row of fluidic control points, and a second pneumatic channel providing suction to a column of fluidic control points, such that the two pneumatic channels together cause fluid to be dispensed from only a single fluid reservoir at a time.
12. The disposable of claim 1 , wherein the first rigid layer further comprises a trench ( 220 ) adjacent to at least one side of the sample analyzing volume ( 500 ), which accepts and distributes fluid flowing from at least one reservoir over a whole side of the the sample analyzing volume ( 500 ).
13. The disposable of claim 12 , wherein the trench ( 220 ) has an asymmetric cross-section.
14. The disposable of claim 1 , wherein the first rigid layer ( 20 ) further comprises a barrier ( 240 ) adjacent to at least one side of the sample analyzing volume ( 500 ), which protrudes into the sample analyzing volume ( 500 ), such that the fluid flows between the barrier ( 240 ) and the support ( 60 ) before entering the sample analyzing volume ( 500 ), and wherein the barrier ( 240 ) has a height of about 200 microns (h 3 ) relative to a surface of the sample analyzing volume ( 500 ).
15. The disposable of claim 11 , wherein a sample analyzing volume ( 500 ) has a height h 1 , a cavity has a depth h 2 , a barrier ( 240 ) has a depth and a biological specimen has a height h 4 and wherein h 1 , h 2 , h 3 and h 4 have ratios of
(0.05 to 0.5)×h 3 =h 4 and/or
(0.1 to 0.5)×h 2 =h 1 and/or
(0.1 to 0.5)×h 1 =h 4 .
16. The disposable of claim 1 , wherein the flexible layer ( 30 ) seals the first rigid layer ( 20 ) against the second rigid layer ( 40 ) and/or the second rigid layer ( 40 ) against the support ( 60 ), and wherein the sample analyzing volume is transparent for light having a wavelength between 200 and 100 nm.
17. The disposable of claim 1 , wherein the plurality of fluid reservoirs ( 22 ) contain a plurality of reagents, of which at least one is an antibody conjugated to a fluorescent molecule.