EP4648608A2

EP4648608A2 - Intubation system and chamber tank

Info

Publication number: EP4648608A2
Application number: EP24706269.8A
Authority: EP
Inventors: Jonathan KRYNITSKY; Thomas T. JONES; Patricia B. GORDON; Kandice TANNER
Original assignee: US Department of Health and Human Services
Current assignee: US Department of Health and Human Services
Priority date: 2023-01-10
Filing date: 2024-01-10
Publication date: 2025-11-19
Also published as: WO2024151752A2; WO2024151752A3

Abstract

An intubation system for directing water to and from an aquatic specimen during the visual observation thereof can include an intubation chamber that facilitates the guided direction of an inflow tube and a plurality of outflow tubes to the specimen. The intubation chamber can be manufactured by an additive manufacturing process to include various inflow and outflow tube channels in a relative complex geometry. An inflow channel guide may be included and can be fitted to the intubation chamber to guide the inflow tube to the intended location of the head of the specimen. The intubation system may also include a brace plate having a chamber opening for the correct positioning of the intubation chamber with respect to the system. A drug delivery system can include a syringe pump and a multiport junction to fluidly introduce a liquid drug to the inflow tube upstream of the intubation chamber.

Description

INTUBATION SYSTEM AND CHAMBER TANK

BACKGROUND

[0001] Zebrafish have become widely used animals in the study and development in human pathology and scientific research, in part because the zebrafish is readily amenable to optical imaging and observation including by microscope. When zebrafish are at the larval stage, they can be easily imaged as they are small with reasonable optical translucence and can be immobilized without deleterious effects. As the zebrafish develops into juvenile and adulthood, it becomes increasingly difficult to immobilize the zebrafish for in vivo imaging over long periods because the zebrafish requires aerated water to continuously pass through its gills.

[0002] To enable the in vivo visual observation of living juvenile and adult zebrafish and similar aquatic specimens, microscope intubation systems have been developed that may hold the immobilized specimen with respect to a vertical microscope. Such microscope intubation systems may include a heated plate having a cavity to receive a petri dish and a lid that can be attached to the heated plate to enclose the cavity. The heated plate may be thermally conductive and can transfer heat from an external source to the petri dish contained in the cavity to maintain the specimen at an appropriate temperature for in vivo observation. The specimen can be placed in the petri dish and the lid may include an aperture to allow visual observation of the specimen. [0003] To provide the specimen with fresh, oxygenated water, one or more tubes or hoses configured for fluid conveyance can be directed into and out of the intubation system via a procedure referred to as intubation. For example, one tube may be an inflow tube supplying water to the petri dish containing the specimen and one or more other tubes may be outflow tubes removing water from the petri dish so that fresh water is continuously circulated through the intubation system. The present disclosure is directed to a system and devices to facilitate the in vivo intubation of a specimen within a microscope incubation system.

BRIEF SUMMARY

[0004] Herein is disclosed an intubation system for the in vivo observation of a zebrafish or similar aquatic specimen with a microscope. The intubation system can include a heated plate that may be heated by an external heating source and a lid that may be attached to enclose the heated plate. A petri dish that may hold the specimen can be placed in a cavity disposed in the heated plate which may be visible through one or more apertures disposed in the heated plate and/or lid. To circulate and supply fresh water to the specimen, one or more inflow hoses or tubes and outflow hoses or tubes can be directed to the intubation system and arranged in fluid communication with the cavity containing the petri dish.

[0005] To direct and locate the inflow tubes and the outflow tubes with respect to the specimen, the intubation system can include an intubation chamber that may be inserted into the petri dish and that may include tube guide channels that receive and guide the inflow and outflow tubes with respect to a chamber cavity containing the specimen. For example, an inflow tube channel can be disposed in the intubation chamber and arranged to direct the inflow tube proximate to the intended location of the head of the specimen within the chamber cavity. A first outflow tube channel and a second outflow tube channel can be located at transversely opposite sides of the chamber cavity so that first and second outflow tubes may be positioned approximately mid-length of the specimen. This arrangement ensures that fresh oxygenated water will flow to the mouth of the specimen, through its gills, and be removed by the rearward located first and second outflow tubes. In an embodiment, the first and second outflow tube channels can be configured to allow for vertical adjustment of the respective first and second outflow tubes received therein to enable selective adjustment of the quantity and depth of water present in the intubation chamber.

[0006] Preferably, the intubation chamber is configured for operative association with a single inflow tube and at least two outflow tubes that function redundantly of each other. Accordingly, the inflow rate of water to the chamber cavity will not exceed the outflow rate even if one of the outflow tubes becomes clogged or fails. Water thus cannot overflow the intubation system and damage an adjacent microscope.

[0007] To manufacture the intubation chamber having the inflow and outflow tube channels, which form a relatively complex geometry, an additive manufacturing process such as three- dimensional printing can be employed. In such an additive manufacturing process, examples of which include stereolithography (“SLA”) and fused deposition modeling (“FDM”), material is added and fused together in successive patterned layers with a computer controlled printer nozzle. In an SLA process, the material may be a photopolymer resin that is selectively cured and solidified by a laser or light sheet. In a FDM process, the material may be a filament that is extruded from a hot nozzle onto a build tray. The material is preferably inert and impermeable to water.

[0008] The intubation system can include additional features to facilitate the arrangement and placement of the inflow and outflow tubes for flow of water to and from the specimen. For example, in an aspect, an inflow tube guide can be included that can be removably fit to the inflow tube channel of the intubation channel to guide and place the inflow tube to the desired location in the chamber cavity. The inflow tube can be detached from the intubation chamber and manipulated to facilitate insertion of the inflow tube into the mouth of the specimen. The inflow tube guide can be manufactured by an additive manufacturing process and can be comprised of a plurality of pipe-like guide legs in a double elbow configuration. In another aspect, the intubation system can include a planar brace plate that can be placed between the heated plate and the lid and that may include a chamber opening to position and brace the intubation chamber with respect to the petri dish. The planar brace plate can also be manufactured by an additive manufacturing process such as, for example, FDM and can include various grooves and similar features to guide and direct the inflow and outflow tubes for the flow of water to and from the intubation chamber.

[0009] In another aspect, a drug delivery system may be used in cooperation with the intubation system to precisely introduce a drug to the flow of oxygenated water to the living aquatic specimen under observation. The drug delivery system can include a multiport junction that can be fluidly interconnected and spliced into an inflow tube running between a source of oxygenated water and the intubation chamber accommodating the specimen. The multiport junction includes an additional port that can connect with a syringe pump for the controlled, and measured delivery of a drug. The multiport junction facilitates intermixing of the drug and oxygenated water upstream of the intubation system prior to delivery to the specimen.

[0010] A possible advantage of the disclosure is that it provides for the continuous flow of oxygenated water and temperature regulation of the intubation chamber for the maintenance of specimen health during prolonged observation. Another possible advantage is that the intubation chamber guides the inflow and outflow tubes to and from the living specimen in an organized arrangement to facilitate visual observation. A related possible advantage the intubation chamber overall enables longitudinal of the specimen. A further possible advantage is that the drug delivery system provides for precisely controlled and continuous introduction of a drug to the living specimen during prolonged or extended observation.

[0011] These and other aspects of the disclosure will be apparent from the following detailed description and the accompanying drawings.

BRIEF DESCRIPTION OF THE DRAWINGS

[0012] Figure 1 is a view of an intubation system and interconnected accessories configured to direct aerated water to and from a specimen in the intubation system for long term observation and microscope imaging.

[0013] Figure 2 is a top plan view of a specimen such as a zebrafish immobilized in the intubation system for observation and imaging.

[0014] Figure 3 is a perspective assembly view of the intubation system configured to receive tubing to direct water to and from the specimen immobilized within the system.

[0015] Figure 4 is a perspective exploded view of the intubation system demonstrating the nested assembly of the internal components.

[0016] Figure 5 is top perspective exploded view of the heated plate and lid of the intubation system that can be attached together to enclose a petri dish containing the specimen.

[0017] Figure 6 is a bottom perspective exploded view of the heated plate and lid of the intubation system.

[0018] Figure 7 is a top perspective view of the intubation chamber of the intubation system showing the structural features thereof that may be manufactured using an additive manufacturing process.

[0019] Figure 8 is a bottom perspective view of the intubation chamber of the intubation system showing the structural features thereof that may be manufactured using an additive manufacturing process.

[0020] Figure 9 is a cross-sectional view of the intubation chamber 9 shown in Figures 7 and 8.

[0021] Figure 10 is a perspective view of the inflow tube guide that can accommodate an inflow tube and that can be secured to the intubation chamber. [0022] Figure 11 is perspective view of the specimen holder that can be received in the chamber cavity of the intubation chamber to secure the inflow tube guide thereto.

[0023] Figure 12 is an exploded perspective view of the intubation tube, the inflow tube guide and the specimen holder with respect to the petri dish showing the assembly thereof. [0024] Figure 13 is an exploded perspective view of the brace assembly for securing and positioning the intubation tube with respect to the intubation system.

[0025] Figure 14 is a perspective view of an embodiment of a specimen tool for retrieving an aquatic specimen and immobilizing the specimen for intubation.

[0026] Figure 15 is a perspective view of another embodiment of a specimen tool configured for a juvenile specimen.

[0027] Figure 16 is a perspective exploded view of an embodiment of a mouth piece assembly that can be inserted into the mouth of an aquatic specimen during intubation.

[0028] Figure 17 is a perspective view of the mouth piece assembly of Figure 17 as assembled.

[0029] Figure 18 is a perspective view of an embodiment of a multiport junction included with a drug delivery system to deliver a drug in liquid form to a living specimen accommodated in the intubation system.

[0030] Figure 19 is a perspective of another embodiment of the multiport junction that may be used with the drug delivery system to deliver a drug in liquid form to a living specimen.

DETAILED DESCRIPTION

[0031] Now referring to the drawings, where whenever possible like reference numbers will refer to like elements, there is illustrated an intubation system 10 and related components and devices for the continuous delivery of aerated water to a specimen accommodated within the intubation system for long term observation including with a microscope. In an embodiment, the intubation system 10 is adapted for placement on the observation stage of a microscope for visual observation and imaging of the specimen therein. The microscope may, for example, be an inverted microscope and may be configured to record images of the specimen from the bottom and/or top of the intubation system 10. In addition to microscopic observation, the intubation system 10 may be used for acoustic imaging, X-rays, MRI, and other observation techniques with respect to the specimen. The specimen may be a zebrafish as is commonly used in the study of human pathology, although various aspects of the disclosure will be applicable to any other suitable living animal that may be visually studied. As used herein, the term long term study may be over any suitable time period including minutes, hours, and days.

[0032] To deliver oxygenated water to the intubation system 10, a fluid pump 12 such as a multi-channel peristaltic pump can be used. A peristaltic pump can include an internal rotor that abuts and rotates against internal hoses or tubing to thereby displace fluid within the hoses. The speed of rotation can be adjusted to change the pumping volume of the fluid pump 12. The fluid pump 12 can include its own reservoir or it may be in fluid communication with an external reservoir. Hollow, flexible, thermoplastic tubing or hosing can be used to fluidly connect the intubation system 10 to the fluid pump 12. The tubing may include at least one inflow tube 14 and one or more outflow tubes 16, 18. In accordance with convention, oxygenated water will flow into the intubation system 10 via the at least one inflow tube 14 and water having passed through the intubation system 10 will be returned to the fluid pump 12 via the first and second outflow tubes 16, 18. The inflow and outflow tubes may be transparent for visual confirmation of fluid flow. Accordingly, the arrangement shown in FIG. l is a closed fluid circuit with water continuously routed between the intubation system and the fluid pump, although in other embodiments, the arrangement can be an open fluid circuit. In an embodiment, the fluid pump 12 can be a four channel pump configured to fluidly communicate with the intubation system 10 via four hoses or tubes.

[0033] In an embodiment, the water delivered by the fluid pump 12 can be oxygenated for maintenance of the living specimen in the intubation system 10 during observation. The oxygenated water may also contain or have added thereto an anesthesia to sedate the specimen in the intubation system 10. The arrangement of the intubation system 10 and fluid pump 12 to deliver oxygenated water with an anesthesia facilitate the prolonged study of the specimen. [0034] In addition to the intubation system 10 and the fluid pump 12, other components may be included in the arrangement to facilitate the study and visual observation of the specimen. For example, to dispense a substance such as, for example, chemicals, nutrients, pathogens, etc. to the specimen in the intubation chamber 10, a syringe pump 20 can be fluidly connected with the inflow tube 14 by a multiport junction 24 located upstream of the intubation chamber 10 as described more fully below. The syringe pump 20 can be configured to deliver measured quantities or dosages of the substance of interest to the specimen.

[0035] The syringe 22 can include a plunger reciprocally movable within a cylindrical tubular barrel and can be fitted with a hypodermic needle tip at one end. The syringe pump 20 is configured to depress the plunger of a syringe 22 to discharge drug in liquid form contained therein. The size of the syringe 22 and the adjustable rate at which the syringe pump 20 depresses the plunger controls and meters the discharge of the drug in liquid form. The syringe pump 20 can be programmable to adjust the speed or rate of the plunger depression and thus the discharge quantity from the syringe 22. The syringe pump 200 can also be programmed to intermittently deliver metered dosages of the drug in liquid form to oxygenated water during prolonged study of the aquatic specimen.

[0036] To evacuate the intubation chamber 10 of fluid, for example, at the termination of the study, an evacuation pump 26 can be included and can be fluidly connected to the intubation chamber via an evacuation tube 28. In the closed fluid circuit arrangement, components can be included to aerate the water circulating there through. In addition, temperature controlled heaters may be included along the fluid circuit to maintain the temperature of the oxygenated water delivered to the specimen. Any other suitable components and devices may be included to aid in the study and observation of the specimen.

[0037] Referring to FIG. 2, there is illustrated an example of a specimen 30 immobilized and accommodated within the intubation system 10. The illustrated specimen 30 may be a zebrafish although as indicated above any suitable aquatic animal can be used with the intubation system 10. The specimen 30 can be centrally located within the structure of the intubation system 10 and can be accommodated within an internal specimen space 32 that is arranged to secure the specimen in a generally immobilized manner during observation while preserving its life. To allow visual observation, imaging, photography and the like of the specimen, the intubation system 10 can include one or more observation apertures or windows 34 enabling viewing into the internal specimen space 32. As described above, the intubation system 10 can include an aperture or observation window 34 enabling viewing into the internal specimen space 32 from either the top or bottom of the intubation system 10. The inflow tube 14 and the outflow tubes 16, 18 to and from the intubation system 10 are arranged so as to not obstruct the observation window 34 and obscure the specimen space 32.

[0038] As can be appreciated, to insert and remove the specimen 30 from the internal specimen space 32, the intubation system 10 can be configured as a multipart assembly allowing access to the internal specimen space. In addition, the intubation system 10 can include various internal channels, passages, grooves and the like to guide at least the inflow tube 14 and the first and second outflow tubes 16, 18 to and from the internal specimen space 32. The first and second outflow tubes 16, 18 can function redundantly of each other such that if one outflow tube fails the second outflow tube will continue to remove water from the intubation system and prevent overflow.

[0039] Referring to FIGS 3, the multipart intubation system 10 can have a generally rectangular shape or other cuboid shape when assembled (FIG. 3) and the multiple components can be configured to nest or fit together in a secured assembly to accommodate the specimen. To facilitate the nested construction of the intubation chamber 10, the individual components can have shapes, profiles, and structural formations that enable them to receive each other and interfit together. Because of the rectangular shape, the intubation system 10 may be a six-sided or six-face structure and may have a bottom face 50, and an oppositely situated top face 52, a first transverse side 54 and an oppositely situated second transverse side 56, and a first lateral end 58 and an oppositely situated second lateral end 60. Consistent with convention, the first and second transverse sides 54, 56 may be dimensionally longer than the first and second lateral ends 58, 60.

[0040] The spatial distance between the bottom face 50 and the top face 52 may be such that the intubation system 10 has a relatively flat rectangular shape. The flat rectangular shape of the intubation system 10 and the short dimension between the bottom and top faces 52 enables the intubation system to be compatible with microscopes from various manufacturers having different configurations and dimensions for the viewing platform. However, in other embodiments, the intubation system 10 can have any other suitable shape (e g., circular) and dimensional size and may include any number of faces or surfaces.

[0041] For reference purposes to facilitate orientation, the direction between the bottom face 50 and the top face 52 can be referred to as the upward/downward direction 70, the direction between the first transverse side 54 and the second transverse side 56 can be referred to as the transverse direction 72, and the direction between the first lateral end 58 and the second later end 60 can be referred to as the lateral direction 74. It will be appreciated that the upward/downward direction 70, the transverse direction 72, and the lateral direction 74 are provided for reference purposes and form no limitation on the scope of the disclosure or claims.

[0042] Referring to FIG. 4 and FIGS. 5 and 6, the external casing 100 of the intubation system 10 is comprised of a heated plate 102 and a lid 104 that, when assembled, enclose a petri dish 106. The heated plate 102 is generally rectangular in shape to correspond to the generally rectangular shape of the intubation chamber 10. To accommodate the petri dish 106, the heated plate 102 includes an internal plate cavity 108 defined in the upper plate face. The plate cavity 108 is generally shaped and dimensioned to correspond to the shape and dimension of the petri dish 106. To allow visual observation of the contents of the petri dish 106 from below, the heated plate 102 can include a plate aperture 110 disposed through the lower plate face.

[0043] To incubate the specimen as necessary to keep it alive, the heated plate 102 can be configured to thermally conduct heat to the petri dished 106 nested in the plate cavity 108. For example, the heated plate 102 can include electrically resistive heating elements disposed within the structure of the heated plate 102. In another embodiment, the heated plate 102 can physically contact external heating elements and is made of a thermally conductive material capable of transferring heat energy to the petri dish 106 accommodated in the plate cavity 108.

[0044] The lid 104 is generally formed as a rectangular frame with four frame edges 112 that outline and define a frame aperture or frame window 114 to allow for visual observation and microscope illumination into the plate cavity 108 from the upper face of the intubation system 10. In the illustrated embodiment, the frame aperture or frame window 114 can be rectangular to correspond with the rectangular shape of the lid 104, but in other embodiments may have any suitable shape. To engage the heated plate 102, the lid 104 can include a rim 116 that depends from the four frame edges 112 and that is dimensioned to surround and secure to the corresponding four sides of the heated plate 102. The lid 104 may held in place under its own weight or may be fastened to the heated plate 102 by any suitable fastening method.

[0045] The petri dish 106 can be a relatively shallow, cup-shaped structure including a circular planar dish bottom 120 which, from the peripheral edge of, extends a cylindrical dish sidewall 122. The planar dish bottom 120 and the cylindrical dish sidewall 122 surround and define an internal dish cavity 124 that can accommodate the specimen. To conform with the shallow shape of the petri dish 106, the diameter of the cylindrical dish sidewall 122 can be larger than its height. The diameter of cylindrical dish sidewall 122 can correspond to the diameter of the plate cavity 108 of the heated plate 102 to securely nest therein. To enable visual observation of the specimen that may be contained therein, the petri dish is made of a transparent material such as glass or transparent plastic. In an embodiment, a thin cover slip can be placed on the inner surface of the circular dish bottom 122 and adhered adjacent thereto by a biocompatible PFPE (perfluoropolyether) grease seal or silicone based resin. The PFPE grease seal may also be disposed to mate the petri dish 106 to the internal plate cavity 108 of the heated plate 100. The grease seal importantly provides a water tight sealed interface between the petri dish and the plate cavity 108, preventing possible water damage to the microscope or imaging system or inadvertent discharge of toxins delivered to the intubation system 10. The petri dish 106 can be a commercial component or can be custom designed to conform in shape with the geometry and dimensions of the internal plate cavity 108 of the heated plate 102.

[0046] Referring to FIG. 4, to facilitate intubation of the specimen, the intubation system 10 can include a chamber assembly 130 configured to nest within the petri dish 106 and regulate the flow of water within the intubation system. The chamber assembly 130 can include an intubation chamber 132, an inflow tube guide 134, and a tube guide holder 136 that can fit together in a mating arrangement. The arrangement of the chamber assembly 130 can selectively direct inflowing oxygenated water to the specimen and can control the removal water from the intubation system 10 in an adjustable manner. When assembled, the interconnected components of the chamber assembly 130 can be placed into the dish cavity 124 of the petri dish 106 that can function as an outer container of the water for the aquatic specimen.

[0047] Referring to FIGS. 7-9, the intubation chamber 132 can have various features and structures formed into the chamber body 140 that may channel the inflow and outflow tubes in a selective arrangement to control the flow of water to and from the specimen. Structurally, the chamber body 140 can include a peripheral chamber wall 142 that can be upright in orientation. The peripheral chamber wall 142 defines a chamber cavity 144 that is shaped and dimensioned to spatially accommodate the specimen, for example, a zebrafish. In an embodiment, the chamber cavity 144 can be generally oblong and can define a first chamber fore-end 146 and a laterally oppositely located second chamber aft-end 148. In use, the chamber fore-end 146 is intended to be proximate with the head of the specimen when it is placed in the intubation chamber 132. To complete the structural outline of the chamber cavity 144, the peripheral chamber wall 142 of the chamber body 140 can include a first chamber side 150 and a transversely opposite second chamber side 152. With respect to the upward/downward direction 70, the chamber body 140 can define a planar upper chamber surface 154 and a lower chamber base 156.

[0048] To mate within the cylindrical sidewall 122 of the petri dish 106, the exterior chamber surface 158 of the peripheral chamber wall 142 can be generally curved at the chamber fore end 146 and the chamber aft-end 148. The distance between the chamber fore-end 146 and the chamber aft-end 148 at the exterior chamber surface 158 can be dimensioned to slidingly abut against the interior of the cylindrical sidewall 122 of the petri dish 106. The exterior chamber surface 158 of the first and second chamber sides 150, 152 can likewise be curved and may have a stepped configuration between the lower chamber base 156 and the upper chamber surface 154. In an embodiment, because of the curved shape, the exterior chamber surface 158 can be generally oval.

[0049] To conform to the shape of the specimen such as a zebrafish, the chamber cavity 144 defined by the peripheral chamber wall 142 can be oblong and may have a generally diamondshape or lozenge-shape. The volume of the chamber cavity 144 can correspond to the physical size of the specimen. The interior chamber surface 159 of the peripheral chamber wall 142 can be smooth and continuous between the upper chamber surface 154 and the lower chamber base 156.

[0050] The chamber body 140 can include structural features to advantageously direct the inflow and outflow tubes with respect to the specimen that may be placed in the intubation chamber 132. For example, to direct the inflow tube proximate to the head of the specimen, an inflow tube channel 160 can be located at the chamber fore-end 148. The inflow tube channel 160 can be formed as a groove disposed into the material of the chamber body 140 and may extend partially along the upper chamber surface 154 and downwardly along the interior chamber surface 159 toward the lower chamber base 156. As described below, the inflow tube channel 160 can securely accommodate the inflow tube guide.

[0051] To selectively channel the first and second outflow tubes, the chamber body 140 can define therein a respective first outflow tube channel 162 and a second outflow tube channel 164. The first and second outflow tube channels 162, 164 can be oppositely located at the respective first chamber side 150 and the opposing second chamber side 152 approximately midway between the chamber fore-end 146 and the chamber aft-end 148. The first outflow tube channel 162 and the second outflow tube channel 164 are thus located transversely opposite each other within the chamber cavity 144. The first and second outflow tube channels 162, 164 can be arranged to direct the respective outflow tubes with respect to the upper planar surface 154 and downwardly with respect to the peripheral chamber wall 142 toward the lower chamber base 156. Locating the first and second outflow tube channels 162, 164 laterally rearward of the inflow tube channel 160 causes the water to flow from the intended location of the head of the specimen rearward and through its gills. Furthermore, locating the first and second outflow tubes channels 162, 164 at the transversely opposing first and second chamber sides 150, 152 restricts the outflow tubes within the chamber cavity 144 and avoids obstructing observation of the aquatic specimen.

[0052] In an embodiment, to enclose the first and second outflow tube channels 162, 164 that accommodate the respective outflow tubes, a first outflow channel boss 166 and a respective second outflow channel boss 168 can be located along the interior chamber surface 159 of the peripheral chamber wall 142. The first and second outflow channel bosses 166, 168 can be generally rounded and can protrude into the chamber cavity 144. The first and second outflow channel bosses 166, 168 can extend from the upper chamber surface 154 toward the lower chamber base 156.

[0053] In an embodiment, to control to the volume of water contained in the chamber cavity 144, the first and second chamber bosses 166, 168 may structurally terminate above the lower chamber base 156. This arrangement enables selective regulation of the height of water contained in the chamber cavity 144. For example, the outflow tubes can be inserted to extend at different distances from the bottom of the first and second chamber bosses 166, 168 and can maintain the water level in the chamber cavity 144 at that established level. [0054] To accommodate an evacuation tube, the chamber body 140 can have an evacuation tube channel 170 disposed therein. The evacuation tube channel 170 can extend transversely along the upper planar surface 154 of the chamber body 140 and can depend downwardly through the structure of the peripheral chamber wall 156. Unlike the first and second outflow tube channels 162, 164, the evacuation tube channel 170 can terminate at the exterior chamber surface 158 of the chamber body 140. Accordingly, when the evacuation tube is received in the evacuation tube channel 170, the evacuation tube can fluidly communicate directly with the dish cavity of the petri dish to evacuate water therefrom.

[0055] The intubation chamber 132 can include other structural features to facilitate prolonged intubation of the specimen. For example, to accommodate one or more probes that can measure water temperature and/or water level in the chamber cavity 144, one or more probe channels 172 can be disposed into the chamber body 140 at one of the first and second chamber sides 150, 152. The probe channels 172 can be disposed within the material of the chamber body 140 between the exterior chamber surface 158 and the interior chamber surface 159 and can extend downwardly within the peripheral chamber wall 142. The probe channels 172 can be circular and smaller in diameter than the first and second outflow tubes 162, 164. Probes such as thermocouples or conductivity sensors, which may be embodied as long fibers or strings having a long, flexible structure, can be inserted through the probe channels 172 to physically contact the water that may be accommodated in the petri dish and make appropriate measurements. To fluidly communicate directly with the petri dish, the probe channel 172 can terminate adjacent to the exterior chamber surface 158 outside the chamber cavity 144.

[0056] In an embodiment, the intubation chamber 132 can be configured with an overflow duct 173 that is disposed in the chamber body 140 and that may extend from the lower chamber base 156 and which is open to the chamber cavity 144 through the interior chamber surface 159. The height at which the overflow duct 173 opens into the chamber cavity 144 may determine or set the quantity and depth of water that can accumulate in the chamber cavity 144. If the water level in the chamber cavity 144 were to rise to the level of the overflow duct 173, the overflow duct would direct the water out to, for example, the petri dish. Probes disposed in the probe channels 172 may detect the overflow water and shut power to the fluid pump preventing additional water from entering the intubation chamber 132. The probes may also activate the evacuation pump to remove water, for example, via the evacuation tube channel 170.

[0057] To situate the intubation chamber 132 within the intubation system 10, in an embodiment, a first chamber support flange 174 can be formed at the chamber fore-end 146 and a respective, oppositely located second chamber support flange 176 can be formed at the chamber aft-end 148. The first chamber support flange 174 and the second chamber support flange 176 can extend in opposite lateral directions from the peripheral chamber wall 142 and can be generally coplanar with the upper chamber surface 154. The first chamber support flange 174 can have a bifurcated structure and the second chamber support flange 176 can be unitary in shape. In an embodiment, to secure the inflow tube guide 134 during assembly, a locating slot 178 can be disposed into the upper chamber surface 154 of the chamber body 140 proximate to the location of the first chamber support flange 174 and the inflow tube channel 160. In an embodiment, one or more circular magnet indentures 179 can be disposed into the first and second chamber support flanges 174, 176 to accommodate correspondingly shaped permanent magnets that may be used for securely locating the intubation chamber 132 within the intubation system as described below.

[0058] In accordance with an aspect of the disclosure, to facilitate manufacturing the complex geometry of the intubation chamber 132, an additive manufacturing process such as SLA can be used. In additive manufacturing, material such asa photopolymer resin can be selectively cured/solidified layer by layer using a laser or light sheet to create a part. Additive manufacturing facilitates the formation of the various tube channels and structural features of the intubation chamber 132. Additive manufacturing also facilitates scaling the size and dimensions of the intubation chamber 132 for different specimens of varying sizes, for example juvenile and adult zebrafish. Additive manufacturing also allows for rapid commercialization of the intubation chamber 132 in desired or custom sizes and arrangements.

[0059] Referring to FIG. 10, to direct the inflow tube proximate to the head of the specimen, the inflow tube guide 134 included with chamber assembly 130 can be structurally formed as a plurality of interconnected pipes shaped to guide the inflow tube with respect to the intubation chamber 132. The pipes that form the inflow tube guide 134 can be circular in shape and can fully or partially enclose the inflow tube. Referring to FIG. 10, in an embodiment, the inflow tube guide 134 is a double elbowed configuration including a first guide leg 180, a second guide leg 182 parallel to the first guide leg 180 and the third guide leg 184 normal to and extending between the first guide leg 180 and the second guide leg 184. The first guide leg 180 is joined to the third guide leg 184 at one end thereof by a first elbow 186 and the second guide leg 182 is joined to the third guide leg 184 at the other end thereof by a second elbow 188.

[0060] Referring to FIG. 4, when connected to the intubation chamber 132, the inflow tube guide 134 is located at the chamber fore-end 146 and can be generally accommodated and nested in the inflow tube channel 160 and can extend from the upper chamber surface 154 to the lower chamber base 156 along the interior chamber surface 159 of the peripheral chamber wall 144. The inflow tube can be inserted through the inflow tube guide 134 and can assume the double elbow shape so that the outlet of the inflow tube is proximate the lower chamber base 156 at the chamber fore-end 146. The outlet of the inflow tube can deliver oxygenated water into the chamber cavity 144 at the chamber fore-end 146 proximate to where the head of the specimen will be located. The inflow tube may be directed from the second guide leg 182 of the inflow tube guide 134 into the mouth of the specimen.

[0061] In an embodiment, to secure the inflow tube guide 134 with the intubation chamber 132, the inflow tube guide 134 can include a first locating arm 190 and a second locating arm 192 that protrude transversely from the first guide leg 180 in opposite directions. The first and second guide legs 190, 192 can be generally rectangular in shape and can be configured to fit within the locating slots 178 disposed in the upper chamber surface 154 proximate to the first chamber support flange 174. To facilitate handling of the inflow tube guide 134, for example, when inserting the inflow tube to the mouth of the specimen, a guide protrusion 194 can protrude from the third guide leg 184. In an embodiment, to facilitate the geometry of the inflow tube guide 134, the part may be manufactured using an additive manufacturing process.

[0062] Referring to FIG. 11, in an embodiment, to hold and locate the specimen with respect to the inflow tube guide 134 and locate the inflow tube guide 134 with respect to the intubation chamber 132 and with respect to the specimen, the chamber assembly 130 can include the specimen holder 136. The specimen holder 136 is a frame-like structure and may have a lower holder body 200 connected to a first upper flange arm 202 and a second upper flange arm 204 by respective first and second holder uprights 206, 208. The lower holder body 200 is generally oriented laterally and may define a holder opening 210 that is designed and dimensioned to surround the specimen. Disposed in the lower holder body 200 can be one or more flow apertures 212 to allow for the flow of water to and from the holder opening 210. The living specimen can be maintained in an immobilized manner in the holder opening 210 of the specimen holder 136 but is otherwise exposed for visual observation without the need for further bracing or other components.

[0063] The first and second holder uprights 206, 208 can be joined at either end of the lower holder body 200 and can extend upright to the first and second upper flange arms 202, 204 respectively. To accommodate the inlet tube guide 134, the first holder upright 206 can be bifurcated or split to define an upright notch 214. When the inlet tube guide 134 is assembled to the tube guide holder 136, the third leg 184 of the inlet tube guide 134 may fit within the tube guide notch 214. The second holder upright 208 may also include a tube guide notch or may be unitary as illustrated. In an embodiment, to facilitate the geometry of the tube guide holder 136, the part may be manufactured using an additive manufacturing process. The additive manufacturing process allows for custom construction of a specimen holder 136 to accommodate specimens of various sizes such as juvenile and adult zebrafish.

[0064] Referring to FIG. 12, the components of the chamber assembly 130 are configured to mate together and secure themselves in place to appropriately direct the inflow tube and the first and second outflow tubes to the desired locations. For example, the intubation chamber 132 can be received in the dish cavity 124 of the petri dish 106 with the chamber fore-end 146 and the chamber aft-end 148 abutting opposite sides of the cylindrical dish sidewall 122 to brace the intubation chamber within the petri dish. In an embodiment, the height of the intubation chamber 132 can dimensionally correspond to the depth of the dish cavity 124 of the petri dish 106, and both can be sized for accommodation within the internal plate cavity 108 of the heated plate 102. The dimensions of the chamber body 140 can correspond to and be adjusted to match the dimensions of the dish cavity 124 of different petri dish 106, which is a feature enabled by the additive manufacturing process.

[0065] In an embodiment, a PFPE (perfluoropolyether) grease seal or a silicone sheet can be used to secure the lower chamber base 156 to the planar dish bottom 120. One or more silicone gaskets may be included and disposed between the exterior surface of the intubation chamber 132 and the cylindrical dish sidewall 122 of the petri dish 106 to secure the two components together.

[0066] The custom petri dish 106 is configured for the intubation chamber 132 such that the intubation chamber 132 can fit within and be sealed with the petri dish 106 by a biocompatible grease to keep water within the chamber cavity 144 of the chamber body 140 during operation. Accordingly, in the event of overflow or leaks (for example, overflow from the chamber cavity 144 due to outflow clog/malfunction) or leaks due to grease seal failure, the custom petri dish can contain overflow and leaks within the petri dish 106 to prevent leaking onto external equipment such as a microscope.

[0067] The inflow tube guide 134 can be mated to the intubation chamber 132 at the chamber fore-end 146 where it can fit within the inflow tube channel 160. The specimen holder 136 can be inserted into the chamber cavity 144 of the intubation chamber 132 to secure the inflow tube guide 134 to the respective peripheral chamber wall 142 and prevent the inflow tube guide from displacing. The first upper flange arm 202 of the specimen holder 134 can be adjacently atop the first chamber support flange 172 and the second upper flange arm 204 can be adjacently atop the second chamber support flange 174.

[0068] Referring to FIG. 4 and FIG. 13, in an embodiment, to securely locate the intubation chamber 132 in the desired position, the intubation system 10 can include a brace assembly 220 having a brace plate 222 and a brace cover 224. The brace plate 222 can be formed as a generally planar plate having an upper plate surface 226 and an opposing lower plate surface 228 and can have a rectangular outline to conform to the rectangular shape of the intubation system 10. To accommodate the intubation chamber 132, a chamber opening 230 can be disposed through the center of the brace plate 222. The brace opening 230 can be oval in shape to conform to the outline of the intubation chamber 132 and can be dimensioned to fit and nest or mate with the intubation chamber. The brace plate 222 thereby braces the intubation chamber 132 against displacement.

[0069] The brace place 222 can include features to register the intubation chamber 132 in proper position. For example, disposed into the upper plate surface 226 can be a pair of first flange notches 232 at one end of the oval shaped chamber opening 230. When the intubation chamber 132 is disposed in the chamber opening 230 the bifurcated first chamber support flange 174 can be received in the pair of first flange notches 232. Similarly, formed in the upper plate surface 226 of the brace plate 222 at the opposite end of the oval shaped chamber opening 230 can be a second flange notch 234 that can correspond in shape with the second chamber support flange 176 which the second flange notch 234 receives. In the embodiment wherein the first and second chamber support flanges 174, 176 of the intubation chamber 132 include magnet indentations 179, permanent magnets therein can be used to secure the first and second support flanges within the first and second flange notches 232, 234.

[0070] To selectively guide the inflow tube and the first and second outflow tubes to the intubation chamber 134 when disposed and nested in the centrally located chamber opening 230, there can be one or more channels or paths in the embodiment of tube grooves disposed in the upper plate surface 226 of the brace plate 222. An advantage of including tube grooves in the brace plate 222 to receive and secure the inflow and outflow tubes is that the tubes may be assembled and appropriately arranged and the specimen may be intubated before the external casing 100 is moved to the microscope for observation.

[0071] For example, to direct the inflow tube to the chamber opening 230, an inflow tube groove 236 can be disposed in the upper plate surface 226 leading from an inflow tube inlet 238 located at the edge of the brace plate 222. To assist in aligning the inflow tube with the inflow tube inlet 238, the brace plate 222 can have an inlet projection 240 that projects from the edge of the brace plate 222 proximate to the inflow tube inlet. In an embodiment, as shown in FIG. 4, to secure the inflow tube, an inflow tube clamp 241 can be complementary in shape with, and can be placed adjacent to, the inlet projection 240. In an embodiment, the inflow tube clamp 241 can include magnet indentations that can receive magnets that can hold the inflow tube clamp 241 fast to the inlet projection 240 via a complementary magnetic arrangement to clamp and secure the inflow tube to the inflow tube inlet 238.

[0072] To direct the first and second outflow tubes, respective first and second outflow tube grooves 242, 244 can be disposed in the upper plate surface 226 leading from opposing sides of the chamber opening 230 to respective first and second outflow tube outlets 246, 248 located at the corresponding and transversely opposite edges of the brace plate 222. The first and second outflow tubes may be secured to the corresponding first and second outflow tube outlets 246, 248 via a slip fit arrangement. In addition, in the embodiments of the intubation system that include an evacuation tube, an evacuation tube groove 250 can be disposed in the upper plate surface 226 leading from the chamber opening 230 to a discharge tube outlet 252 at the edge of the brace plate 222. The evacuation tube can also be secured to the evacuation tube outlet 252 via a slip fit arrangement.

[0073] The inflow tube groove 236 and the outflow tube grooves 242, 244 provide a visual indication of the proper arrangement of the tubes with respect to the intubation chamber and the specimen therein. The inflow and outflow tubes may be removable from the inflow tube groove 236 and outflow tube grooves 242, 244 respectively to allow for disassembly of the system after use.

[0074] To accommodate the brace cover 224, there can be disposed into the upper surface 226 of the brace plate 222 a cover frame 254. The brace cover 224 may be generally rectangular in shape, smaller in dimension than the brace plate 222, and the cover frame 254 can have a complementary shape to situate the brace cover 224. When placed in the cover frame 254, the brace cover 224 can be located partially over the intubation chamber 132 disposed in the chamber opening 230 of the brace plate 222 and prevent any displacement of the intubation chamber and the inflow and outflow tubes directed thereto by the tube grooves in the brace plate 222. To enable visual observation into the intubation chamber 132 from above, the brace cover 224 can have a cover aperture 256 disposed therein.

[0075] The brace plate 222 and the brace cover 224 can also be manufactured by an additive manufacturing process to allow for different configurations and arrangements of the inflow tube groove 236 and the outflow tube grooves 242, 244. For example, the size of the grooves may be configured to accommodate inflow and outflow tubes of varying sizes to correspond to different species of specimens or specimen sizes.

[0076] In the embodiment wherein the first and second chamber support flanges 174, 176 of the intubation chamber 132 include magnet indentations 179, the magnets included therein can be used to hold the brace cover 224 over the intubation chamber 132. In an embodiment, the brace cover 224 may also include magnet indentations 258 to accommodate permanent magnets for holding the brace cover 224 to the brace plate 222 that may include complementary magnets disposed therein. [0077] The brace plate 222 and the brace cover 224 can sandwich the first and second chamber support flanges 174, 176 of the intubation chamber 132 and the first and second upper flange arms 202, 204 of the specimen holder 134 to stabilize the system and the specimen. The inflow and outflow tubes can be held securely by the brace plate 222 and brace cover 224 and the specimen holder 134 holds the specimen steady after intubation allowing the assembly to be securely moved without harm to the specimen, for example, to a microscope or other imaging system after the specimen is intubated.

[0078] In accordance with additional aspects of disclosure, there is disclosed in FIG. 14 a specimen tool 300 that can be used to retrieve an aquatic specimen such as a zebrafish for intubation. The specimen tool 300 can include an elongated handle 302 having a first handle leg 304 and a second handle leg 306 that are joined together at a bend 308 so that the handle 302 has a bent configuration. Formed at the end of the first handle leg 304 is a scoop 310 that may have a dished or cup-like shape. The scoop 310 can extend orthogonally from the end of a first handle leg 306 of the handle 302 and can be curved with respect to the plane of the first handle leg. A handle slot 312 can be disposed at the location where the scoop 310 and the first handle leg 306 are joined. The specimen tool 300 can be manufactured by an additive manufacturing process such as SLA or FDM.

[0079] In use, the specimen tool 300 can be inserted in an aquarium tank containing aquatic specimens such as zebrafish. The specimens may be anesthetized by an anesthesia introduced to the aquarium water. The specimen can be guided onto the dish-shape scoop 310 and can be partially held tail first in the handle slot 310. The captured specimen can be thereafter removed from the tank and is generally immobilized and stabilized by the specimen tool 300 to intubate the specimen with an inflow tube before being place in the intubation system. The scoop 310 can include one or more scoop apertures 314 to allow for water to drain from the specimen after removal form the tank.

[0080] In an embodiment, the specimen tool 300 can be provided in different sizes depending upon the size of the specimen. For example, the specimen tool 300 may be intended for an adult zebrafish with the scoop 302 and the handle slot 306 correspondingly sized. Referring to FIG. 15, there is illustrated another example of the specimen tool 310 that may be sized for a smaller specimen such as a juvenile zebrafish which are typically smaller in size than adult zebrafish. Similar to the first example, the small specimen tool 320 can have a bent handle 322 including first and second handle legs 324, 326 joined at a bend 328 and can include a dishshaped scoop 330 that extends orthogonally from the end of first handle leg 324. To receive the tail of the specimen, a handle slot 332 can be disposed proximate to where the scoop 330 is joined to the first handle leg 324. The present example of the specimen tool 320 can also be made from an additive manufacturing process.

[0081] Referring to FIGS. 16 and 17, there is illustrated an embodiment of a mouth piece assembly 340 for insertion into the mouth of an aquatic specimen during intubation. The mouth piece assembly 340 can be a multiple part assembly that can be fit into the lumen at the end of the inflow tube 342 and that may be made from a plurality of conduits 344, 346, 348, and 350 that can be plugged together. The plurality of conduits 344 - 354 can be generally cylindrical tubes of varying diameters and can be made of a thermoplastic material. The internal and external diameter of the plurality of tubular conduits 344 - 354 can be sized to allow successive conduits to be inserted and fitted together as shown in FIG. 17. The plurality of conduits 344, 346, 348, 350, 352, and 354 may have decreasing diameters to restrict the flow of water from the inflow tube 342 to a quantity that the specimen can safely intake. In an embodiment, one of the plurality of conduits 344 - 354 may be sized and configured as an adult conduit 350 intended for intubation into the mouth of an adult specimen and another of the plurality of conduits may be sized and configured as a juvenile conduit 354 intended for intubation with a smaller sized mouth of a juvenile specimen. The juvenile conduit 354 can have a smaller diameter than the adult conduit 350 and thereby further reduces the flow of water from the inflow tube 304.

[0082] Referring to FIGS. 18 and 19, in conjunction with FIG. 1, the intubation system 10 can be used in cooperation with a drug delivery system to precisely administer a drug to the living specimen. The drug delivery system shown in FIG. 1 can include the syringe pump 20 and the multiport junction 24 that fluidly connects the syringe 22, which may contain a drug, embodied as a liquid, to the inflow tube 14 from the fluid pump 12 upstream of the intubation system 10. The drug may be in a liquid phase or solute dissolved into a solution. The multiport junction 24 enables the introduction of the drug as a liquid to oxygenated water delivered from the fluid pump 12 so that the liquids intermix upstream of the specimen accommodated in the intubation system 10. The drug delivery system allows for the delivery and experimentation with different drugs and substances using the intubation system.

[0083] Referring to FIG. 18, in an embodiment, the multiport junction 22 can be a three-port Y-j unction 400 including a first fluid port 402 connected with the inflow tube 14 from the fluid pump, a second fluid port 404 that fluidly connects with a continuation of the inflow tube 14 to the intubation chamber, and a third fluid port 406 that fluidly connects with the drug delivery system. In possible embodiments, the multiport junction 22 can have different numbers of fluid ports to enable interaction with multiple drug sources such as multiple syringes. The body of the three-port Y-junction 400 can be made from a transparent material to allow observation of the introduction and mixing of the drug to the oxygenated water.

[0084] The third fluid port 406 can be configured for connection with the needle tip from the syringe 22 coupled with the syringe pump 20 of the drug delivery system. In an embodiment, the third fluid port 406 may include a self-sealing membrane 408 that may be made of a resilient, elastomeric material. The self-sealing membrane 408 can be penetrated by the needle tip of the syringe for the introduction of the drug in liquid form into the three-port Y-junction 400. The self-sealing characteristic of the self-sealing membrane 408 allows for interchangeability of different syringes containing different drugs during the observation and study of the specimen. The small diameter of the needle tip cannula of syringe prevents backflow of fluid after introduction to the multiport junction 22.

[0085] Referring to FIG. 19, in an embodiment, the multiport junction 24 can be a microfluidic junction 500 configured for introduction of the drug at microfluidic quantities. The microfluidic junction 500 can include a first microfluidic fitting 502, a second microfluidic fitting 504 and third microfluidic fitting 506 arranged by a plastic junction body 508. The plastic junction body 508 may be transparent to allow visual observation of the introduction of the drug into the inflowing oxygenated water.

[0086] The microfluidic fittings 502, 504, 506 are configured to couple with appropriate capillary tubing and the discharge orifices 510 of the microfluidic fittings are arranged to adjacently interface with each other within the junction body 510. The small diameters of the discharge orifices 510 enable precise control of micro-quantities of the drug in liquid form to the oxygenated water delivered to the specimen while preventing backflow of the drug in liquid form that would interfere with measurements during the clinical study.

[0087] In an embodiment, the drug delivery system can be configured to deliver a mixture of oxygenated water and the drug in liquid form to multiple specimens simultaneously. For example, the second fluid port 404 of the three-port Y-junction 400, or the second microfluidic fitting 504 of the mi croflui die junction 500, can be connected to a tube of an appropriate diameter that is divided or split into multiple parallel branch tubes downstream of the multiport junction 22 to fluidly communicate with a plurality of intubation systems 10 or similar systems to accommodate the living specimens. Individual specimens can be located in separate intubation chambers by a divided setup with watertight dividers to isolate the inflowing water and the specimens and prevent cross-talk.

[0088] All references, including publications, patent applications, and patents, cited herein are hereby incorporated by reference to the same extent as if each reference were individually and specifically indicated to be incorporated by reference and were set forth in its entirety herein. [0089] The use of the terms “a” and “an” and “the” and “at least one” and similar referents in the context of describing the invention (especially in the context of the following claims) are to be construed to cover both the singular and the plural, unless otherwise indicated herein or clearly contradicted by context. The use of the term “at least one” followed by a list of one or more items (for example, “at least one of A and B”) is to be construed to mean one item selected from the listed items (A or B) or any combination of two or more of the listed items (A and B), unless otherwise indicated herein or clearly contradicted by context. The terms “comprising,” “having,” “including,” and “containing” are to be construed as open-ended terms (i.e., meaning “including, but not limited to,”) unless otherwise noted. Recitation of ranges of values herein are merely intended to serve as a shorthand method of referring individually to each separate value falling within the range, unless otherwise indicated herein, and each separate value is incorporated into the specification as if it were individually recited herein. All methods described herein can be performed in any suitable order unless otherwise indicated herein or otherwise clearly contradicted by context. The use of any and all examples, or exemplary language (e.g., “such as”) provided herein, is intended merely to better illuminate the invention and does not pose a limitation on the scope of the invention unless otherwise claimed. No language in the specification should be construed as indicating any non-claimed element as essential to the practice of the invention.

[0090] Preferred embodiments of this invention are described herein, including the best mode known to the inventors for carrying out the invention. Variations of those preferred embodiments may become apparent to those of ordinary skill in the art upon reading the foregoing description. The inventors expect skilled artisans to employ such variations as appropriate, and the inventors intend for the invention to be practiced otherwise than as specifically described herein. Accordingly, this invention includes all modifications and equivalents of the subject matter recited in the claims appended hereto as permitted by applicable law. Moreover, any combination of the above-described elements in all possible variations thereof is encompassed by the invention unless otherwise indicated herein or otherwise clearly contradicted by context.

Claims

CLAIMS:

1. An intubation system for intubation of an aquatic specimen comprising: a heated plate including an internal plate cavity for accommodating a petri dish; a lid adapted to engage the heated plate and cover the internal plate cavity, the lid including a frame window allowing for visual observation of the internal plate cavity; an intubation chamber adapted for placement in the internal plate cavity, the intubation chamber including a chamber cavity disposed therein, the intubation chamber including an inflow tube channel to direct an inflow tube to the chamber cavity and first outflow tube channel and a second outflow tube channel to direct a first outflow tube and a second outflow tube respectively with the chamber cavity.

2. The intubation system of claim 1, wherein the chamber cavity is defined by a chamber fore-end, a chamber aft-end, a first chamber side and an opposing second chamber side.

3. The intubation system of claim 2, wherein the inflow tube channel is disposed at the chamber fore-end.

4. The intubation system of claim 3, wherein the first outflow tube channel is disposed at the first chamber side and the second outflow channel is disposed at the second chamber side.

5. The intubation system of claim 4, wherein the intubation system includes a first outflow channel boss formed at the first lateral side defining the first outflow tube channel and a second outflow channel boss formed at the second lateral side defining the second outflow tube channel.

6. The intubation system of claim 5, wherein the intubation chamber includes an upper chamber surface and a lower chamber base, the first outflow tube boss and the second outflow tube boss extending from the upper chamber surface downwardly toward the lower chamber base.

7. The intubation system of claim 6, wherein the first outflow tube boss and the second outflow tube boss terminate above the lower chamber base.

8. The intubation system of claim 1, further comprising an inflow tube guide adapted to fit within the inflow tube channel of the intubation chamber.

9. The intubation system of claim 8, wherein the inflow tube guide includes first guide leg, a second guide leg, and a third guide leg joined together in a double elbow configuration.

10. The intubation system of claim 9, wherein the intubation chamber includes an upper chamber surface and a lower chamber base, the first guide leg of the inflow tube guide adjacent the upper chamber surface and the second guide leg of the inflow tube guide disposed proximate when the inflow tube guide is fitted to the inflow tube channel of the intubation chamber.

11. The intubation system of claim 10, further comprising a tube guide holder configured for insertion into the chamber cavity to secure the tube guide in the tube guide channel of the intubation chamber.

12. The intubation system of claim 11, wherein the tube guide holder includes a lower holder body, a first upper flange arm, and a second upper flange arm.

13. The intubation system of claim 9, wherein the inflow tube guide is configured to direct the inflow tube to a mouth of the aquatic specimen.

14. The intubation system of claim 1, wherein the intubation chamber further includes an evacuation tube channel disposed therein.

15. The intubation system of claim 13, wherein the intubation chamber includes an upper chamber surface and a lower chamber base and the evacuation tube channel extends between the upper chamber surface and the lower chamber base.

16. The intubation system of claim 1, wherein the intubation chamber include one or more probe channels disposed therein to accommodate one or more probes.

17. An intubation system for the intubation and study of an aquatic specimen comprising: an intubation chamber including a peripheral chamber wall defining a chamber cavity for receiving the aquatic specimen, the intubation chamber further including an inflow tube channel to direct an inflow tube to the chamber cavity and an outflow tube channel to direct an outflow tube from the chamber cavity; a fluid pump fluidly connected to the intubation chamber by the inflow tube to direct oxygenated water to the chamber cavity; and a drug delivery sub-system fluidly connected to the inflow tube by a multi-port junction to introduce a drug to the oxygenated water upstream of the intubation chamber.

18. The intubation system of claim 17, wherein the multi-port junction is a three-port Y- j unction.

19. The intubation system of claim 18, wherein the drug delivery sub-system includes a syringe pump coupled with a syringe.

20. The intubation system of claim 19, wherein the multi-port junction include a self-sealing injection port fluidly connected with the syringe.

21. The intubation system of claim 20, wherein the fluid pump is also fluidly connected to intubation chamber by the outflow tube.

22. A method of observing a living aquatic specimen comprising: placing the living aquatic specimen in an chamber cavity of an intubation chamber; directing water that is oxygenated from a fluid pump to the chamber cavity though an inflow tube fluidly connecting the fluid pump and the intubation chamber; introducing a drug from a drug delivery system to the oxygenated water through a multiportjunction fluidly connecting the drug delivery system and the inflow tube upstream of the intubation chamber.

23. The method of claim 22, wherein the multi-port junction is a three-port Y-junction with a first port fluidly connected to the fluid pump, a second port fluidly connected to the intubation chamber, and a third port fluidly connected to the drug delivery system.

24. The method of claim 23, wherein the drug delivery system is a syringe pump coupled with a syringe.

25. The method of claim 24, wherein the third port is a self-sealing injection port having a self-sealing membrane for penetration with a needle tip of the syringe.

26. The method of claim 25, further comprising the step of intubating the living aquatic specimen by inserting the inflow tube to its mouth.

27. The method of claim 26, further comprising the step of removing the water from the chamber cavity with an outflow tube in fluid communication with the fluid pump.

28. A drug delivery system for administration of a drug to a living aquatic specimen comprising: a syringe pump coupled with a syringe for the controllable discharge of a drug; an inflow tube for delivery water that has been oxygenate to the living aquatic specimen; and a multiport junction fluidly connecting a needle tip of the syringe with the inflow tube to the living aquatic specimen.

29. The drug delivery system of claim 28, wherein the multiport junction includes a selfsealing injection port with a self-sealing membrane for penetration by the needle tip of the syringe.

30. The drug delivery system of claim 28, wherein the syringe pump is programmable to control a quantity of the drug introduced to the inflow tube from the syringe.