Systems and methods for in vivo nanotoxicity testing
View Patent ↗Systems and methods are provided for comprehensive in vivo nanotoxicity testing, as well as fabrication methods of chips used for nanotoxicity testing. A three-dimensional (3D)-printed, automated microfluidic chip can be configured for in vivo nanotoxicity testing, and the chip can allow for multi-route (e.g., waterborne and/or foodborne) exposure of marine organisms to small materials (e.g., plastics such as microplastics and/or nanoplastics (NPLs), or metals), providing real-time, in situ monitoring of bioaccumulation and oxygen consumption.
1 . A system chip configured for in vivo nanotoxicity testing, the system comprising:
a microfluidic chip that comprises:
a first chamber;
a first inlet channel connected to the first chamber;
a first outlet channel connected to the first chamber;
a second chamber;
a second inlet channel connected to the second chamber;
a second outlet channel connected to the second chamber; and
a valve disposed between the first chamber and the second chamber;
a control channel connected to the valve;
a first pump configured to control the valve via the control channel;
a second pump configured to supply a first fluid to the first chamber via the first inlet channel;
a third pump configured to supply a second fluid to the second chamber via the second inlet channel;
a chemical sensor connected to the first chamber;
a film heater configured to regulate a temperature of within at least one of the first chamber and the second chamber; and
a controller connected to the film heater and configured to control the film heater, the chemical sensor being an optical oxygen sensor,
the optical oxygen sensor being configured to measure in real-time a dissolved oxygen concentration of the first fluid in the first chamber,
the optical oxygen sensor comprising an oxygen molecule-sensitive dye, an optical fiber, and an oxygen meter,
the first pump being a syringe pressure pump,
the first chamber comprising a first intake port configured to intake a first biological sample,
the second chamber comprising a second intake port configured to intake a second biological sample,
the first pump, the second pump, and the third pump all being controlled via an automated flow control,
the first inlet channel comprising a first plurality of micropillars where the first inlet channel meets an entrance to the first chamber,
the second inlet channel comprising a second plurality of micropillars where the second inlet channel meets at an entrance to the second chamber,
the valve being a normally open valve that is configured to be closed during at least a portion of in vivo nanotoxicity testing, and
the microfluidic chip being a monolithic structure such that the first chamber, the first inlet channel, the first outlet channel, the second chamber, the second inlet channel, the second outlet channel, and the valve are all part of the monolithic structure.
2 . A method for in vivo nanotoxicity testing, the method comprising:
providing the system according to claim 1 ;
closing the valve;
providing the first fluid to the first chamber via the first inlet;
providing the second fluid to the second chamber via the second inlet, where at least one of the first fluid and the second fluid comprises nanoparticles (NPs);
providing a first biological sample to the first chamber;
providing a second biological sample to the second chamber;
capturing microscopic images of the first biological sample and the second biological sample; and
measuring in real-time a dissolved oxygen concentration of the first fluid in the first chamber.
3 . The method according to claim 2 , the first fluid being salt water,
the second fluid being salt water, and
the NPs being nanoplastics (NPLs).
4 . The system according to claim 1 , further comprising a fluorescence microscope disposed proximate to the microfluidic chip and configured to capture microscopic images of a sample within the microfluidic chip during in vivo nanotoxicity testing.
5 . A method for in vivo nanotoxicity testing, the method comprising:
providing the system according to claim 4 ;
closing the valve;
providing the first fluid to the first chamber via the first inlet;
providing the second fluid to the second chamber via the second inlet, where at least one of the first fluid and the second fluid comprises nanoparticles (NPs);
providing a first biological sample to the first chamber;
providing a second biological sample to the second chamber;
capturing microscopic images of the first biological sample and the second biological sample using the fluorescence microscope; and
measuring in real-time a dissolved oxygen concentration of the first fluid in the first chamber.
6 . The method according to claim 5 , the first fluid being salt water,
the second fluid being salt water, and
the NPs being nanoplastics (NPLs).
7 . The system according to claim 1 , the controller being a proportional-integral-derivative (PID) controller.
8 . A method for in vivo nanotoxicity testing, the method comprising:
providing the system according to claim 7 ;
closing the valve;
providing the first fluid to the first chamber via the first inlet;
providing the second fluid to the second chamber via the second inlet, where at least one of the first fluid and the second fluid comprises nanoparticles (NPs);
providing a first biological sample to the first chamber;
providing a second biological sample to the second chamber;
capturing microscopic images of the first biological sample and the second biological sample; and
measuring in real-time a dissolved oxygen concentration of the first fluid in the first chamber.
9 . The method according to claim 8 , the first fluid being salt water,
the second fluid being salt water, and
the NPs being nanoplastics (NPLs).
10 . The system according to claim 1 , the syringe pressure pump comprising a first syringe pump and a pressure sensor.
11 . A method for in vivo nanotoxicity testing, the method comprising:
providing the system according to claim 10 ;
closing the valve;
providing the first fluid to the first chamber via the first inlet;
providing the second fluid to the second chamber via the second inlet, where at least one of the first fluid and the second fluid comprises nanoparticles (NPs);
providing a first biological sample to the first chamber;
providing a second biological sample to the second chamber;
capturing microscopic images of the first biological sample and the second biological sample; and
measuring in real-time a dissolved oxygen concentration of the first fluid in the first chamber.
12 . The method according to claim 11 , the first fluid being salt water,
the second fluid being salt water, and
the NPs being nanoplastics (NPLs).
13 . The system according to claim 1 , the second pump comprising a second syringe pump.
14 . A method for in vivo nanotoxicity testing, the method comprising:
providing the system according to claim 13 ;
closing the valve;
providing the first fluid to the first chamber via the first inlet;
providing the second fluid to the second chamber via the second inlet, where at least one of the first fluid and the second fluid comprises nanoparticles (NPs);
providing a first biological sample to the first chamber;
providing a second biological sample to the second chamber;
capturing microscopic images of the first biological sample and the second biological sample; and
measuring in real-time a dissolved oxygen concentration of the first fluid in the first chamber.
15 . The method according to claim 14 , the first fluid being salt water,
the second fluid being salt water, and
the NPs being nanoplastics (NPLs).
16 . The system according to claim 13 , the third pump comprising a third syringe pump.
17 . A method for in vivo nanotoxicity testing, the method comprising:
providing the system according to claim 16 ;
closing the valve;
providing the first fluid to the first chamber via the first inlet;
providing the second fluid to the second chamber via the second inlet, where at least one of the first fluid and the second fluid comprises nanoparticles (NPs);
providing a first biological sample to the first chamber;
providing a second biological sample to the second chamber;
capturing microscopic images of the first biological sample and the second biological sample; and
measuring in real-time a dissolved oxygen concentration of the first fluid in the first chamber.
18 . The method according to claim 17 , the first fluid being salt water,
the second fluid being salt water, and
the NPs being nanoplastics (NPLs).
19 . The system according to claim 1 , the first chamber being larger than the second chamber.
20 . A method for in vivo nanotoxicity testing, the method comprising:
providing the system according to claim 1 ;
closing the valve;
providing the first fluid to the first chamber via the first inlet;
providing the second fluid to the second chamber via the second inlet, where at least one of the first fluid and the second fluid comprises nanoparticles (NPs);
providing a first biological sample to the first chamber;
providing a second biological sample to the second chamber;
capturing microscopic images of the first biological sample and the second biological sample; and
measuring in real-time a dissolved oxygen concentration of the first fluid in the first chamber,
the first fluid being salt water,
the second fluid being salt water, and
the NPs being nanoplastics (NPLs).