Cartridge and system for manipulating samples in liquid droplets

A system (40) for liquid droplet manipulation, the system comprising a substrate (42) with an electrode array (20) and a central control unit (43) for controlling the selection of individual electrodes (44) of the electrode array (20) and for providing the electrodes (44) with individual voltage pulses for manipulating liquid droplets (23) by electrowetting, wherein the system (40) is configured to receive on top of the electrodes (44) a cartridge (1) with a working film (10) and at least one piercing element (13) for manipulating samples in liquid droplets with an electrode array (20) when the working film (10) of the cartridge (1) is placed on said electrode array (20), wherein the cartridge (1) comprises:

• a) a body (2,2′,2″) that comprises an upper surface (3), a lower surface (4), and a number of wells (5) configured to hold therein reagents (6) or samples (6′), each well (5) comprising a top opening (72), and a bottom opening (73) for releasing a liquid from the well (5);
• b) a piercable bottom structure (8) impermeable to liquids and configured to seal at least one of the bottom openings (73) of the wells (5);
• c) a working film (10) located below the lower surface (4) of the body (2,2′,2″), the working film (10) being impermeable to liquids and comprising a hydrophobic upper surface (11);
• d) a peripheral spacer (9) located below the lower surface (4) of the body (2,2′,2″) and connecting the working film (10) to the body (2,2′,2″);
• and
• e) a gap (12) between the lower surface (4) of the body (2,2′,2″) and the hydrophobic upper surface (11) of the working film (10), the gap (12) being defined by the peripheral spacer (9);
• wherein the cartridge (1) further comprises
• f) at least one top piercing system (60), each located within at least one of the wells (5) for releasing a reagent or sample (6,6′) from said at least one well (5) into the clap (12);
• wherein each top piercing system (60) comprises a piston (61) and a piercing element (13), the piston (61) being configured to be movable within said well (5) while providing a seal between the piston (61) and the inner wall of the well (5), and the piercing element (13) being configured as a thorn (63) located at a front side (74) of the piston (61) and being configured to pierce the piercable bottom structure (8) for releasing a reagent or sample (6,6′) from said at least one well (5) into the gap (12) upon moving the piston (61) within the well (5) toward its bottom opening (73).

A method of releasing a reagent or sample (6,6′) from a well (5) of a cartridge (1) with a working film (10) and at least one piercing element (13) for manipulating samples in liquid droplets with an electrode array (20) when the working film (10) of the cartridge (1) is placed on said electrode array (20), wherein the method comprises the following steps:

• providing a cartridge (1) which comprises:
• a) a body (2,2′,2″) that comprises an upper surface (3), a lower surface (4), and a number of wells (5) configured to hold therein reagents (6) or samples (6′), each well (5) comprising a top opening (72), and a bottom opening (73) for releasing a liquid from the well (5);
• b) a piercable bottom structure (8) impermeable to liquids and configured to seal at least one of the bottom openings (73) of the wells (5);
• c) a working film (10) located below the lower surface (4) of the body (2,2′,2″), the working film (10) being impermeable to liquids and comprising a hydrophobic upper surface (11);
• d) a peripheral spacer (9) located below the lower surface (4) of the body (2,2′,2″) and connecting the working film (10) to the body (2,2′,2″);
• e) a gap (12) between the lower surface (4) of the body (2,2′,2″) and the hydrophobic upper surface (11) of the working film (10), the gap (12) being defined by the peripheral spacer (9);
• and
• f) at least one top piercing system (60), each located within at least one of the wells (5) for releasing a reagent or sample (6,6′) from said at least one well (5) into the gap (12);
• wherein each top piercing system (60) comprises a piston (61) and a piercing element (13), the piercing element (13) being configured as a thorn (63) located at a front side (74) of the piston (61);
• moving the piston (61) within said well (5) toward its bottom opening (73) while providing a seal between the piston (61) and the inner wall of the well (5);
• piercing the piercable bottom structure (8) with the thorn (63);
• and
• releasing a reagent or sample (6,6′) from said at least one well (5) into the gap (12) upon moving the piston (61) within the well (5).

A system configured to perform a method of releasing a reagent or sample (6,6′) from a well (5) of a cartridge (1) with a working film (10) and at least one piercing element (13) for manipulating samples in liquid droplets with an electrode array (20) when the working film (10) of the cartridge (1) is placed on said electrode array (20), wherein the method comprises the following steps:

• providing a cartridge (1) which comprises:
• a) a body (2,2′,2″) that comprises an upper surface (3), a lower surface (4), and a number of wells (5) configured to hold therein reagents (6) or samples (6′), each well (5) comprising a top opening (72), and a bottom opening (73) for releasing a liquid from the well (5);
• b) a piercable bottom structure (8) impermeable to liquids and configured to seal at least one of the bottom openings (73) of the wells (5);
• c) a working film (10) located below the lower surface (4) of the body (2,2′,2″), the working film (10) being impermeable to liquids and comprising a hydrophobic upper surface (11);
• d) a peripheral spacer (9) located below the lower surface (4) of the body (2,2′,2″) and connecting the working film (10) to the body (2,2′,2″);
• e) a gap (12) between the lower surface (4) of the body (2,2′,2″) and the hydrophobic upper surface (11) of the working film (10), the gap (12) being defined by the peripheral spacer (9);
• and
• f) at least one top piercing system (60), each located within at least one of the wells (5) for releasing a reagent or sample (6,6′) from said at least one well (5) into the gap (12);
• wherein each top piercing system (60) comprises a piston (61) and a piercing element (13), the piercing element (13) being configured as a thorn (63) located at a front side (74) of the piston (61);
• moving the piston (61) within said well (5) toward its bottom opening (73) while providing a seal between the piston (61) and the inner wall of the well (5);
• piercing the piercable bottom structure (8) with the thorn (63);
• and
• releasing a reagent or sample (6,6′) from said at least one well (5) into the gap (12) upon moving the piston (61) within the well (5).