Cassette assembly
A cassette assembly ( 1 ) comprises a cover ( 100 ), two cassette halves ( 200 A, 200 B) and a slider ( 300 ) comprising multiple test chambers ( 302 ). The cassette halves ( 200 A, 200 B) comprise waste tanks ( 230 ) in fluid connection with reservoirs ( 240 ), prefilled in one of the cassette halves ( 200 A, 200 B) with test agents. The particular design of the cassette halves ( 200 A, 200 B) enable forming predefined 5 volumes of liquid to achieve predefined concentrations of the test agents in the reservoirs ( 240 ) in one of the cassette halves ( 200 A, 200 B) and liquid in the reservoirs ( 240 ) in the other of the cassette halves ( 200 A, 200 B). Gradients of the test agents can then be established over the multiple test chambers ( 302 ).
1 . A cassette assembly comprising:
a cover;
a first cassette half;
a second cassette half; and
a slider, wherein
the cover is configured to be positioned onto the first cassette half and the second cassette half and keep the first cassette half and second cassette half together with the slider sandwiched between the first cassette half and the second cassette half;
the slider comprises N≥2 test chambers in the form of through holes through the slider;
each of the first cassette half and the second cassette half comprises:
N waste tanks connected in series and separated by respective walls; and
an excessive liquid tank in fluid connection with the waste tanks by a vertical outlet channel and by an air valve;
an inlet port of the first cassette half is configured to receive a liquid to sequentially fill each waste tank and a head space defined above the waste tanks; and
the air valves are configured to be opened to drain liquid in the head spaces into the excessive liquid tanks through the vertical outlet channels and to form a separation of equal volumes of liquid in the waste tanks;
wherein each of the first cassette half and the second cassette half comprises N reservoirs;
each reservoir in one of the first cassette half and the second cassette half comprises a respective test agent and each corresponding reservoir in the other of the first cassette half and the second cassette half lacks the respective test agent; and
each waste tank is in fluid connection with a respective reservoir through a liquid channel extending from the waste tank turning at the slider and continuing to the reservoir.
2 . The cassette assembly according to claim 1 , wherein the air valves are configured to present a higher flow resistance to the liquid as compared to the vertical outlet channels to prevent liquid from entering the excessive liquid tanks through the air valves during filling of the waste tanks.
3 . The cassette assembly according to claim 1 , wherein
each of the first cassette half and the second cassette half comprises a connection port configured to be aligned with respective openings in the cover; and
the air valves are configured to be opened by a first overpressure pulse applied through the openings in the cover.
4 . The cassette assembly according to claim 1 , wherein
the air valves are configured to be opened by a suction of air from the excessive liquid tanks through the air valves and into the head spaces; and
the suction of air is caused by gravitational forces acting on liquid in the vertical outlet channels or by absorption of liquid in the vertical outlet channels by absorbent material comprised in the excessive liquid tanks.
5 . The cassette assembly according to claim 1 , wherein
the air valves are in the form of channels interconnecting the head spaces and the excessive liquid tanks;
the cross-sectional dimensions of the vertical outlet channels relative to the channels of the air valves are configured to create an under pressure by gravitation and capillary force that is sufficiently large to draw excess liquid into the excessive liquid tank but not sufficiently large to open the air valves during filling of the waste tanks.
6 . The cassette assembly according to claim 1 , wherein
each of the first cassette half and the second cassette half comprises a connection port configured to be aligned with respective openings in the cover; and
the equal volumes of liquid in the waste tanks are configured to be pressed out from the waste tanks and into the reservoirs via the liquid channels by application of a second over pressure pulse through the openings in the cover.
7 . The cassette assembly according to claim 6 , wherein
each liquid channel comprises a restrictive filter or slit; and
the second over pressure pulse is configured to open the restrictive filters or slits to allow emptying of the equal volumes of liquid in the waste tanks into the reservoirs.
8 . The cassette assembly according to claim 7 , wherein the restrictive filters or slits are configured to prevent liquid from passing the restrictive filters or slits during filling of the waste tanks but enable liquid to flow through the liquid channels past the restrictive filters or slits once an opening pressure of the restrictive filters or slits has been overcome by application of the second over pressure pulse through the openings in the cover.
9 . The cassette assembly according to claim 1 , wherein
each of the first cassette half and the second cassette half comprises a connection port configured to be aligned with respective openings in the cover;
membranes are arranged on top of the reservoirs and configured to allow air but not liquid to pass through the membranes; and
the membranes are configured to be opened by application of an under pressure pulse at the openings in the cover to introduce a volume of air between the liquid in the reservoirs and the membranes.
10 . The cassette assembly according to claim 1 , wherein
the slider is movable relative to the first cassette half and the second cassette half between a sample filling position and a flow position; and
the liquid channels are in fluid connection with the test chambers in the slider in the flow position but not in the sample filling position.
11 . The cassette assembly according to claim 10 , wherein
each of the first cassette half and the second cassette half comprises a connection port configured to be aligned with respective openings in the cover;
liquid in the reservoirs are configured to flow through the liquid channels into the waste tanks by application of an under pressure at the openings in the cover when the slider is in the flow position; and
one side of a respective three dimensional (3D) culture matrix in the test chambers is exposed to liquid comprising a test agent and the other side of the respective 3D culture matrix is exposed to liquid lacking the test agent to establish a linear concentration gradient of the test agent over the respective 3D culture matrix.
12 . The cassette assembly according to claim 1 , wherein
the slider comprises a first through hole in connection with a first end of the slider and a second through hole in connection with a second end of the slider;
each of the first cassette half and the second cassette half comprises:
a circumferential channel having a first opening configured to be aligned with the first through hole and a second opening configured to be aligned with the second through hole;
a serpentine channel interconnecting the circumferential channel and the waste tanks;
the inlet port of the first cassette half is in fluid connection with the circumferential channel; and
the serpentine channels are configured to present a higher flow resistance to the liquid as compared to the circumferential channels to enable filling of the circumferential channels prior to entering of the liquid into the waste tanks through the serpentine channels.
13 . The cassette assembly according to claim 12 , wherein
each of the first cassette half and the second cassette half comprises a vertical inlet channel in fluid connection at its lower end with the serpentine channel and in fluid connection with the waste tanks in its upper end.
14 . The cassette assembly according to claim 1 , wherein
the slider is movable relative to the first cassette half and the second cassette half between a sample filling position and a flow position;
one of the first cassette half and the second cassette half comprises an inlet port configured to receive a biological sample comprising a gel suspension that can be polymerized into three dimensional (3D) culture matrices in the test chambers in the slider when the slider is in the sample filling position; and
each of the first cassette half and the second cassette half comprises a channel system forming, together with the test chambers in the slider, a meander pattern and configured to sequentially fill the test chambers with the biological sample received at the inlet port.
15 . The cassette assembly according to claim 14 wherein
the slider is configured to be moved relative to the first cassette half and the second cassette half from the sample filling position into the flow position once the gel suspension has been polymerized into the 3D culture matrices in the test chambers; and
the relative movement between the slider and the first cassette half and the second cassette half is configured to cut the polymerized biological sample to form well defined sides of the 3D culture matrices and to align the sides of the 3D culture matrices with liquid openings in front walls of the first cassette half and the second cassette half.
16 . The cassette assembly according to claim 14 , wherein
each of the first cassette half and the second cassette half comprises N reservoirs;
each reservoir in one of the first cassette half and the second cassette half comprises a respective test agent and each corresponding reservoir in the other of the first cassette half and the second cassette half lacks the respective test agent;
each waste tank is in fluid connection with a respective reservoir through a liquid channel extending from the waste tank turning at the slider and continuing to the reservoir;
each of the first cassette half and the second cassette half comprises a front wall configured to face the slider; and
each front wall comprises N pairs of openings comprising a respective upper opening in fluid connection with a respective liquid channel and a respective lower opening forming part of the channel system.
17 . The cassette assembly according to claim 16 , further comprising gaskets or seals arranged in the front walls to circumferentially enclose the openings.
18 . The cassette assembly according to claim 1 , wherein the cover comprises a lid and supporting structures extending from the lid and supporting a slider cover comprising windows configured to be aligned with the test chambers in the slider to provide visual access through the slider cover into the test chambers.
19 . A system for determining a response of microorganisms to test agents comprising:
a cassette assembly according to claim 1 ;
an analysis instrument configured to take at least one image of three dimensional (3D) culture matrices in the test chamber, wherein the 3D culture matrices are formed by polymerization of a biological sample comprising the microorganisms and a gel suspension; and
a processor configured to determine a response of the microorganisms to test agents based on the at least one image of the 3D culture matrices.
20 . A cassette assembly comprising:
a cover;
a first cassette half;
a second cassette half; and
a slider, wherein
the cover is configured to be positioned onto the first cassette half and the second cassette half and keep the first cassette half and second cassette half together with the slider sandwiched between the first cassette half and the second cassette half;
the slider comprises N≥2 test chambers in the form of through holes through the slider;
each of the first cassette half and the second cassette half comprises:
N waste tanks connected in series and separated by respective walls; and
an excessive liquid tank in fluid connection with the waste tanks by a vertical outlet channel and by an air valve;
an inlet port of the first cassette half is configured to receive a liquid to sequentially fill each waste tank and a head space defined above the waste tanks; and
the air valves are configured to be opened to drain liquid in the head spaces into the excessive liquid tanks through the vertical outlet channels and to form a separation of equal volumes of liquid in the waste tanks;
wherein the slider comprises a first through hole in connection with a first end of the slider and a second through hole in connection with a second end of the slider;
each of the first cassette half and the second cassette half comprises:
a circumferential channel having a first opening configured to be aligned with the first through hole and a second opening configured to be aligned with the second through hole;
a serpentine channel interconnecting the circumferential channel and the waste tanks;
the inlet port of the first cassette half is in fluid connection with the circumferential channel; and
the serpentine channels are configured to present a higher flow resistance to the liquid as compared to the circumferential channels to enable filling of the circumferential channels prior to entering of the liquid into the waste tanks through the serpentine channels.
21 . The cassette assembly according to claim 20 , wherein
each of the first cassette half and the second cassette half comprises a vertical inlet channel in fluid connection at its lower end with the serpentine channel and in fluid connection with the waste tanks in its upper end.
22 . A cassette assembly comprising:
a cover;
a first cassette half;
a second cassette half; and
a slider, wherein
the cover is configured to be positioned onto the first cassette half and the second cassette half and keep the first cassette half and second cassette half together with the slider sandwiched between the first cassette half and the second cassette half;
the slider comprises N≥2 test chambers in the form of through holes through the slider;
each of the first cassette half and the second cassette half comprises:
N waste tanks connected in series and separated by respective walls; and
an excessive liquid tank in fluid connection with the waste tanks by a vertical outlet channel and by an air valve;
an inlet port of the first cassette half is configured to receive a liquid to sequentially fill each waste tank and a head space defined above the waste tanks; and
the air valves are configured to be opened to drain liquid in the head spaces into the excessive liquid tanks through the vertical outlet channels and to form a separation of equal volumes of liquid in the waste tanks;
wherein the slider is movable relative to the first cassette half and the second cassette half between a sample filling position and a flow position;
one of the first cassette half and the second cassette half comprises an inlet port configured to receive a biological sample comprising a gel suspension that can be polymerized into three dimensional (3D) culture matrices in the test chambers in the slider when the slider is in the sample filling position; and
each of the first cassette half and the second cassette half comprises a channel system forming, together with the test chambers in the slider, a meander pattern and configured to sequentially fill the test chambers with the biological sample received at the inlet port.
23 . The cassette assembly according to claim 22 wherein
the slider is configured to be moved relative to the first cassette half and the second cassette half from the sample filling position into the flow position once the gel suspension has been polymerized into the 3D culture matrices in the test chambers; and
the relative movement between the slider and the first cassette half and the second cassette half is configured to cut the polymerized biological sample to form well defined sides of the 3D culture matrices and to align the sides of the 3D culture matrices with liquid openings in front walls of the first cassette half and the second cassette half.
24 . The cassette assembly according to claim 22 , wherein
each of the first cassette half and the second cassette half comprises N reservoirs;
each reservoir in one of the first cassette half and the second cassette half comprises a respective test agent and each corresponding reservoir in the other of the first cassette half and the second cassette half lacks the respective test agent;
each waste tank is in fluid connection with a respective reservoir through a liquid channel extending from the waste tank turning at the slider and continuing to the reservoir;
each of the first cassette half and the second cassette half comprises a front wall configured to face the slider; and
each front wall comprises N pairs of openings comprising a respective upper opening in fluid connection with a respective liquid channel and a respective lower opening forming part of the channel system.
25 . The cassette assembly according to claim 24 , further comprising gaskets or seals arranged in the front walls to circumferentially enclose the openings.