LIQUID RESERVOIRS FOR MAXIMIZING REAGENT RECOVERY

Described herein are various liquid reservoirs that may be used in a laboratory setting. The liquid reservoirs minimize waste of pipetted liquid sample or reagents and may be used, e.g., in combination with a multi-well plate such as in NGS processes. Other reservoirs include those configured for compatibility with centrifuge rotors or multichannel pipettes with 12 or more channels.

BACKGROUND

The ability to efficiently pipette liquids can become a limiting factor in virtually any chemistry or life sciences laboratory. Whether due to the limited availability of precious samples that have taken weeks (or months) to produce, or the use of expensive reagents—such as enzymes, antibodies and probes—there is a real cost to every microliter wasted. The value of samples or reagents needs to be balanced against productivity and throughput, because using a single channel pipette capable of accessing the last few microliters of a reagent or sample is a laborious and time-consuming process. Multichannel pipettes allow faster and more reproducible assay setup, but must be used in combination with reagent reservoirs such that the tips of all of the individual pipettes simultaneously draws from the reagent reservoir. This can be a drawback, because the high dead volumes of conventional reservoirs increase the cost of experiments.

Next generation sequencing (NGS) is one example where the high cost of NGS reagents demands low waste. In addition, the time required to precisely perform all of the necessary low volume pipetting steps adds to the cost of analysis. Multichannel electronic pipettes have the potential to significantly reduce the time required for library preparation in microplates or tube racks, but conventional reagent reservoirs tend to be very wasteful due to their large dead volumes. There is a need to provide commercially-available low-cost low-waste solutions to improve laboratory workflow and to decrease waste of precious reagent or sample.

The present disclosure provides a variety of solutions to the aforementioned challenges as well as compatibility with many of the tools used in NGS, thereby simplifying workflow.

SUMMARY

Provided herein are various liquid reservoirs that may be used, e.g., in a laboratory setting to minimize loss of a reagent, sample, or other liquid. In any embodiment, a liquid reservoir comprises a walled perimeter formed of at least one wall segment and a bottom segment, defining a liquid space where liquid may be contained. In various embodiment, the liquid reservoir may be sized to receive and/or connect to a multi-well plate.

DETAILED DESCRIPTION

The present disclosure provides various liquid reservoirs for minimizing loss of a liquid and/or for use with large multi-channel pipettes, such as those having 12 or more channels. In general, the liquid reservoirs comprise a walled perimeter and a floor portion attached thereto, defining an interior space therein for containing a liquid. The liquid may be a reagent, a sample, or any other liquid used in a laboratory setting. The liquid reservoirs comprise various beneficial design characteristics which will now be described with respect to example embodiments below.

Single Low Point Reservoir

In one embodiment, the present disclosure provides a liquid reservoir comprising a single low point where liquid may pool for maximal recovery of the liquid. As such, a liquid reservoir will generally have a walled perimeter comprising at least one wall segment attached to a floor portion, thereby defining liquid space. The liquid reservoir comprising the single low point (herein “single low point reservoir”) may be substantially rigid in structure and be sized for compatibility with systems typically used in combination with a liquid reservoir, such as a well plate (e.g., 96 well micro-plate, polymerase chain reaction (PCR) plate). As used herein, “well plate” and “microplate” are used interchangeably and refer to a plate with a plurality of wells that can hold a volume of liquid and are arranged in a regular (e.g., rectangular) pattern. Well plates are available in many shapes and sizes, depending on any given laboratory application.FIG.1shows one embodiment of a single low point reservoir100which comprises a walled perimeter102formed of wall segments104,106,108,110, the walled perimeter having a top edge112, a bottom edge114, and a floor portion116joined to the walled perimeter102to form a liquid space120configured to hold a volume of liquid. The floor portion116has at least one indentation118with a lowest point140provided therein. The first wall segment104, second wall segment,106, third wall segment,108, and fourth wall segment110define a generally rectangular walled perimeter102wherein the first wall segment104and third wall segment108are parallel and the second wall segment106and fourth wall segment110are parallel.

On each of the first wall segment104and third wall segment108, there is a longitudinal support projection122extending into the liquid space120. On each of the first wall segment104and third wall segment108, there are two longitudinal securing projections138located above the longitudinal support projection122and extending to the top edge114of the walled perimeter102. As used herein, “longitudinal” is used to refer to the largest dimension of the liquid reservoir. As used herein, “lateral” is used to refer to refer to the direction orthogonal to the longitudinal direction and parallel to a plane formed by the intersection of the floor portion116with each wall segment104,106,108,110.

The second wall segment106and fourth wall segment110each comprise at least one lateral support projection130. On each of the second wall segment106and fourth wall segment110, there are two lateral securing projections134located above the lateral support projection130and extending to the top edge114of the walled perimeter102. Longitudinal and lateral securing projections (134,138) are optional, but may be included to prevent plate movement during centrifugation and/or to hold an inverted microplate more securely. The embodiment depicted inFIG.1comprises two securing projections on each wall segment, however, any number of securing projections may be used, such as 0 (absent), 1, 2, 3, 4, or more, on each side. In any embodiment, some wall segments may include one or more securing projections while other wall segments are absent securing projections.

Additionally, the number of longitudinal and lateral support projections122,130are not particularly limited to one on each wall segment, as depicted inFIG.1. Each wall segment may comprise, e.g., 0, 1, 2, 3 or more support projections on each wall segment. Further, one or more support projections may be present on only one pair of parallel wall segments (e.g., first and third or second and fourth) while the other pair of parallel wall segments are absent support projections. Additionally, the number of supporting projections may be different on each wall segment. The shape of the longitudinal and lateral supporting projections122,130inFIG.1are such that each wall segment indents inwards towards the liquid space120to form a void or cavity in the wall with respect to the walled perimeter. However, in any embodiment described herein, including those described below, an indentation may not form the projection, but rather the projection may be formed as a separate element attached to the wall segment.

FIG.2, where like numbers represent like elements, depicts a lateral cross section that passes through the single low point140of the single low point reservoir100. The at least one longitudinal support projection122comprises a top surface132positioned at a nesting distance128below the top edge114of the walled perimeter102.FIG.3, where like numbers represent like elements, depicts a longitudinal cross section that passes through the single low point140of the single low point reservoir100. The at least one lateral support projection130comprises a top surface126positioned at the nesting distance128below the top edge114of the walled perimeter102. Nesting distance, as used herein, is the distance between the top of a supporting projection and the top edge of the reservoir and may be utilized, e.g., to support an inverted microplate that is nested in the top edge of the liquid reservoir.

While indentation118and lowest point140are depicted inFIGS.2and3as located substantially in the center of the floor portion116, the indentation comprising the lowest point may be location at any point in the floor portion116.

A bottom view of the single low point reservoir100shown inFIG.1is provided inFIG.4, where like numbers represent identical elements. Indentations124,131corresponding to the longitudinal support projections122and lateral support projections130(fromFIGS.1-3) can be seen. The floor portion116connects to the walled perimeter102at a location117between the top edge112and the bottom edge114. The floor portion116is shaped to provide the lowest point118, which is structurally stabilized within the void surrounding it with support structures136. While four support structures136are shown inFIG.4, any number of support structures136may be used, such as 2, 3, 5, 6, 7, or 8. The support structures136provide mechanical strength to the overall reservoir structure. In any embodiment, however, the void formed between the floor portion116and the bottom edge114of the walled perimeter102need not be void and could instead be solid or partially solid (e.g., partially filled or comprise additional support structures).

In any embodiment, the indentation118may be shaped substantially as shown inFIG.1, where the lowest point140is at the apex of an inverted pyramid having four faces. In any embodiment, such an inverted pyramid may have three faces or more than four faces, such as 5, 6, 7, or 8 faces. Alternatively, and in any embodiment, the inverted pyramid may be partially or fully conical. The intersection of the indentation118with the floor portion116may be angular, as shown inFIG.1or may substantially curved to avoid an edge at the junction thereof.FIG.1depicts an inverted pyramid indentation118that is nested within the floor portion116. In any embodiment, there may be two or more successively nesting indentations, such an inverted pyramid nested within a larger inverted pyramid, nested within the floor portion116.

Divided Reservoir

In another embodiment, the present disclosure provides a divided liquid reservoir comprising multiple subdivisions, each comprising a low point, that compatible for use with multiple liquids (e.g., different reagents) where liquid may pool for maximal recovery of the liquid contained in each subdivision. A divided liquid reservoir will generally have a walled perimeter comprising at least one wall segment attached to a floor portion, thereby defining a liquid space which may be divided into at least two subdivisions by one or more partition walls. A liquid reservoir comprising multiple subdivisions, each with a single low point (herein “divided reservoir”) may be substantially rigid in structure and be sized for compatibility with systems typically used in combination with a liquid reservoir, such as a well plate (e.g., 96 well micro-plate, polymerase chain reaction (PCR) plate).FIG.5shows one embodiment of a divided reservoir200which comprises a walled perimeter202formed of wall segments204,206,208,210, the walled perimeter having a top edge212, a bottom edge214, and a floor portion216joined to the walled perimeter202. Partition wall203, oriented perpendicular to one first and third wall segments204,208form two liquid spaces220,221. The floor portion216,217of each liquid space220,221has an indentation218,219with a lowest point240,241(not visible) provided therein. The first wall segment204, second wall segment,206, third wall segment,208, and fourth wall segment210define a generally rectangular walled perimeter202wherein the first wall segment204and third wall segment208are parallel and the second wall segment206and fourth wall segment210are parallel.

FIG.6depicts a top view of the divided liquid reservoir ofFIG.5, where all like numbers represent like elements. InFIG.6, both indentations218,219are visible.

FIG.5depicts a divided reservoir with two liquid spaces each with an indentation and lowest point, however, in any embodiment there may be 4, 8, 12, or more liquid spaces, each with a lowest point. Further, the partition wall203depicted inFIG.5is optional. In embodiments lacking a partition wall, a multiple low-point reservoir is created with a single liquid space with multiple indentations and low points contained therein.

Longitudinal Trough Liquid Reservoir

In another embodiment, the present disclosure provides a liquid reservoir for minimizing loss of a liquid while also providing compatibility with a larger multi-channel pipette, e.g., a multi-channel pipette having at least twelve (12) channels. The liquid reservoir is capable of holding a volume of liquid and comprises a trough spanning a longitudinal axis of the reservoir for pooling of a liquid. Such a liquid reservoir will generally have a walled perimeter comprising at least one wall segment and a floor portion attached thereto, defining liquid space. A liquid reservoir comprising a longitudinal trough (herein “longitudinal trough reservoir”) may be substantially rigid in structure and be sized for compatibility with systems typically used in combination with a liquid reservoir, such as a microplate (e.g., a 96 well plate or a PCR plate).FIG.7shows one embodiment of a longitudinal trough reservoir300which comprises a walled perimeter302formed of wall segments304,306,308,310, the walled perimeter having a top edge312and a bottom edge314and a floor portion316joined to the walled perimeter302to form a liquid space320configured to hold a volume of liquid. The floor portion316has at least one longitudinal trough indentation318that spans a length of the longitudinal trough reservoir300with a substantially two-dimensional bottom340to minimize liquid loss therein.

In any embodiment, the trough indentation may be shaped substantially as shown inFIG.7, having where the lowest point340is at the apex of an inverted triangular cross-section. The intersection of the indentation318with the floor portion316may be angular, as shown inFIG.7or may substantially curved to avoid an edge at the junction thereof.

Longitudinal trough reservoir300comprises two lateral support projections330on each of the second wall segment306and fourth wall segment310and two lateral securing projections334located above the lateral support projection330and extending to the top edge314of the walled perimeter302. Longitudinal trough reservoir300comprises two longitudinal support projections322on each of the first wall segment304and third wall segment308and two longitudinal securing projections338located above the longitudinal support projection322and extending to the top edge314of the walled perimeter302. Again, the securing projections (334,338) are optional, but serve to prevent plate movement during centrifugation and hold an inverted well or PCR plate more securely. The embodiment depicted inFIG.7comprises two securing projections on each wall segment, however, any number of securing projections may be used, such as 0 (absent), 1, 2, 3, 4, or more, on each side. In any embodiment, some wall segments may include one or more securing projections while other wall segments are absent securing projections.

InFIG.8(where like numbers represent like elements), which depicts the centermost lateral cross-sectional view of the longitudinal trough reservoir300, the at least one longitudinal support projection322has a top surface325positioned at a nesting distance328below the top edge314of the walled perimeter302. The lateral cross section of the at least one trough indentation318has a low point340, which forms the bottom of the at least one trough indentation318.

InFIG.9(where like numbers represent like elements), which depicts the centermost longitudinal cross section of the longitudinal trough reservoir300, the at least one lateral support projection330comprises a top surface332positioned at the nesting distance328below the top edge314of the walled perimeter302.

A bottom view of the longitudinal reservoir300shown inFIG.7is provided inFIG.10, where like numbers represent identical elements. Indentations324,331corresponding to the longitudinal support projections322and lateral support projections330can be seen. The floor portion316connects to the walled perimeter302at a location between the top edge312and the bottom edge314and the floor portion316is shaped to provide a trough indentation318which is structurally secured with two support structures336. The support structures336provide mechanical strength to the overall reservoir structure.

Rounded Bottom Edge

In any embodiment, the bottom edge, being substantially rectangular in each ofFIGS.1-4and6-10, may have rounded corners as shown inFIG.11(and inFIG.5). Advantageously, these rounded corners may provide compatibility with various common laboratory instruments such as a centrifuge or rotor compatible with deep well plates (which are typically about 40 mm to about 45 mm tall), such as the Eppendorf™ Rotor for Benchtop Centrifuge or Aerosol-tight deepwell plate Rotor A-2-DWP-ATI, sold by Fisher Scientific, which can be used with Eppendorf™ centrifuges.

FIG.11depicts a longitudinal trough rounded bottom reservoir400with a longitudinal trough indentation418similar to that shown inFIGS.7-10and comprising two longitudinal support projections422on each of the first wall segment404and third wall segment408and two longitudinal securing projections438located above the longitudinal support projection422and extending to the top edge414of the walled perimeter402. Longitudinal trough rounded bottom reservoir400comprises two lateral support projections430on each of a second wall segment406and fourth wall segment410and two lateral securing projections434located above the lateral support projection330and extending to the top edge414of the walled perimeter402. A rounded plane442carved each corner444of the walled perimeter402defines rounded bottom corners446. The rounded plane has a length441and is defined by a radius of curvature and an arc length (not shown inFIG.11). Again, the securing projections (434,438) are optional, but serve to prevent plate movement during centrifugation and hold an inverted well or PCR plate more securely. The embodiment depicted inFIG.11comprises two securing projections on each wall segment, however, any number of securing projections may be used, such as 0 (absent), 1, 2, 3, 4, or more, on each side. In any embodiment, some wall segments may include one or more securing projections while other wall segments are absent securing projections.

A bottom view of the longitudinal trough rounded bottom reservoir400shown inFIG.11is provided inFIG.12, where like numbers represent identical elements as described inFIG.11. Indentations424,431corresponding to the longitudinal support projections422and lateral support projections430can be seen. The floor portion416connects to the walled perimeter402at a location between the top edge412and the bottom edge414and the floor portion416is shaped to provide a trough indentation418which is structurally secured with two support structures436. The support structures436provide mechanical strength to the overall reservoir structure but reduce the amount of construction material required (for example, the void area between the floor portion416and the bottom edge414of the walled perimeter402could be solid).

WhileFIG.11depicts a longitudinal trough reservoir with a rounded bottom reservoir, a single low point reservoir may also have a rounded bottom, such as shown inFIG.13.FIG.13depicts a single low point reservoir500which comprises a walled perimeter502formed of wall segments504,506,508,510, the walled perimeter having a top edge512and a bottom edge514and a floor portion516joined to the walled perimeter502to form a liquid space520configured to hold a volume of liquid. The floor portion516has at least one indentation518with a lowest point540contained therein. The first wall segment504, second wall segment,506, third wall segment,508, and fourth wall segment510define the generally rectangular walled perimeter502wherein the first wall segment504and third wall segment508are parallel and the second wall segment506and fourth wall segment510are parallel.

On each of the first wall segment504and third wall segment508, there is a longitudinal support projection522extending into the liquid space520. On each of the first wall segment504and third wall segment508, there are two longitudinal securing projections538located above the longitudinal support projection522and extending to the top edge514of the walled perimeter502.

The second wall segment506and fourth wall segment510each comprise at least one lateral support projection530. On each of the second wall segment506and fourth wall segment510, there are two lateral securing projections534located above the lateral support projection530and extending to the top edge514of the walled perimeter502. Again, the securing projections (534,538) are optional, but serve to prevent plate movement during centrifugation and hold an inverted well or PCR plate more securely. The embodiment depicted inFIG.13comprises two securing projections on each wall segment, however any number of securing projections may be used, such as 0 (absent), 1, 2, 3, 4, or more, on each side. In any embodiment, some wall segments may include one or more securing projections while other wall segments are absent securing projections.

A rounded plane542carved into each bottom corner544of the walled perimeter502defines rounded bottom corners546. The rounded plane has a length541and is defined by a radius of curvature and an arc length (not shown inFIG.13).

A bottom view of the single low point rounded bottom reservoir500shown inFIG.13is provided inFIG.14, where like numbers represent identical elements as described inFIG.13. Indentations524,531corresponding to the longitudinal support projections and lateral support projections, respectively can be seen. The floor portion516connects to the walled perimeter502at a location between the top edge and the bottom edge. The floor portion516is shaped to provide an indentation518which is structurally secured with support structures536. The support structures536provide mechanical strength to the overall reservoir structure.

FIG.15depicts a general cross-section of many of the embodiments disclosed herein, having an indentation618in a floor portion616with a low point640(representing either a single low point or the lowest indentation of a trough). The size of the various elements shown, such as the projections622and the size of the indentation618may be described by various fill lines, represented by dotted lines inFIG.15. Fill line 1610, corresponding to the top of the indentation618may be a first distance above the low point640. Fill line 2620, corresponding to where the wall604and the floor portion616meet, may be a second distance above the low point640. Fill line 3630, corresponding to the top surface of the supporting projection622may be a third distance above the low point630. For example, in any embodiment, the first distance may be about 2.5 mm to about 3 mm, such as about 2.92 mm. In any embodiment, the second distance may be about 12 mm to about 15 mm, such as about 14 mm. In any embodiment, the third distance may be about 25 mm to about 30 mm, such as about 28 mm.

The size of the various elements shown, such as the projections622and the size of the indentation618may additionally or alternatively be described by various fill lines corresponding to volumes of fluid that are contained within the reservoir. Fill line 1610, corresponding to the top of the indentation618may correspond to a first volume. Fill line 2620, corresponding to where the wall604and the floor portion616meet, may correspond to a second volume. Fill line 3630, corresponding to the top surface of the supporting projection622may correspond to a third volume. For example, in any embodiment, the first volume may be about 1 mL to about 3 mL, such as about 1.15 mL. In any embodiment, the second volume may be about 55 mL to about 60 mL, such as about 59 mL. In any embodiment, the third volume may be about 195 mL to about 200 mL, such as about 197 mL.

Functional Design Elements

Advantageously, the various design elements of the reservoirs described above enable compatibility with laboratory equipment often used therewith, such as centrifuges, waste receptacles, well plates (e.g., 6-, 12-, 24-, 48-, 96-, 384-, or 1536-well microplates, including PCR microplates which may be non-skirted or skirted). For example, a liquid reservoir as described herein may be sized to allow nesting of an inverted well plate and/or PCR plate containing a liquid reagent in the liquid reservoir, such that the wells face the liquid space but are supported above the floor portion by the supporting projections. Therefore, the nesting distance, which is the distance between the top of a supporting projection and the top edge of the reservoir, may be dictated to correspond to a feature common to many well plates. A 96-well plate700is shown inFIGS.16a-e, having 96 wells702and a flange748, which can be better seen inFIG.16b.FIG.16bdepicts a partial cross-sectional view where the edge704of the well plate700comprises a flange748. As particularly shown inFIGS.16c-e, upon inversion of the well plate700and insertion into the top of a liquid reservoir800comprising at least one support projection on each wall, the flange748may rest on the top surface of each support projection to provide nesting of the well plate in the reservoir.FIG.16edepicts a cross-sectional view of how the flange748of inverted well plate700may rest on at least one support projection of each wall of liquid reservoir800. As such, the nesting height of the support projections, as described inFIGS.1-14, may correspond to a flange height728of a well plate to be used therewith. In any embodiment, a flange may be absent (e.g., non-skirted or semi-skirted PCR plate) or be about 0.1 mm to about 5 mm in height. Typically, flange heights on standardized well plates are 2 mm to 2.5 mm. In deep well plates, the flange height may range from 2.5 mm to 8 mm. Therefore, the nesting distance, in any embodiment, may be about 0.1 mm to about 5 mm, such as about 2.0 mm, about 2.5 mm, about 3 mm, about 4 mm, about 5 mm, about 6 mm, or approximately the size of a flange on a compatible well plate. In embodiments compatible with non-skirted or semi-skirted PCR plates, a supporting projection may be any size, simply serving to provide support to the overturned PCR plate nested within the reservoir. For example, in any embodiment where a flange is absent, an upper surface of a well plate740may rest upon the support projections (e.g., lateral support projection(s)830), as described inFIGS.1-14, preferably where the upper surface contacts the support projection at a location that does not overlap with any wells in the well plate.

The well-plate inFIG.16aalso is characterized by a length l and a width w, which may be any value compatible or suitable with a workflow or instrument in a laboratory. For example, well plates are typically about 100 mm to about 150 mm in length and about 70 mm to about 100 mm wide. Generally, the size of a well plate is standardized across manufacturers for compatibility across a wide variety of uses. For example, the length of well plates available from ThermoFisher and Grainger have a length of 127.76 mm and a width of 85.48 mm. For a snug fit, the inside length of the reservoir (dictated by the length of the first and third wall segments and subtracting the wall thickness therefrom) may be slightly smaller than the outside length of the well plate for a snug fit. For example, in any embodiment, the inside length of a reservoir, as described herein, may be about 0.5 to about 1.5 mm smaller than a compatible well plate, such as about 0.5 mm to about 1 mm, about 1 mm to about 1.5 mm, or about 0.75 mm to about 1.25 mm. The exact length will depend on the flexibility of the construction material of the reservoir, which will be discussed further below. For a well plate that is about 127 mm to about 128 mm long, a suitable inside length may be about 126 mm to about 128 mm. Likewise, the inside width of the reservoir (dictated by the length of the second and fourth wall segments and subtracting the wall thickness therefrom) may be slightly smaller than the outside width of the well plate for a snug fit. For example, in any embodiment, the inside width of a reservoir, as described herein, may be about 0.5 mm to about 1.5 mm smaller than a compatible well plate, such as about 0.5 mm to about 1 mm, about 1 mm to about 1.5 mm, or about 0.75 mm to about 1.25 mm. The exact width will depend on the flexibility of the construction material of the reservoir, which will be discussed further below. For a well plate that is about 85.5 mm long, a suitable inside length may be about 84 mm to about 85 mm, such as about 84.5 mm. A plurality of securing projections, if included and as described above, may further aid in providing a snug fit between the reservoir and an inverted well plate. As such, each securing projection may independently project from its corresponding wall at a distance of about 0.5 mm to about 1.5 mm.

The height of a liquid reservoir is not particularly limited in function, except in any respect related to compatibility with other laboratory instrumentation that will be used therewith. For example, many centrifuges are limited in the height of an object that may be safely contained therein.

In any embodiment, a voided bottom such as shown inFIGS.1-12reduces the amount of construction materials of the reservoir, but also enables compatibility and interoperability with various other devices common in the laboratory and designed to improve workflow, such as, but not limited to, reagent dispensers, liquid waste removers, and adaptors that enable thermal and mechanical motion control of the reservoir.

A liquid reservoir, as described herein, may be made of any material, and may be selected based on an intended use. For example, a liquid reservoir may be manufactured with materials that are resistant to degradation by water, solvents, and other frequently used reagents as well as high temperature (e.g., for sterilization) and have high mechanical strength (e.g., for use in a centrifuge). The surface that will contact the liquid (e.g., the surface of the floor portion and the inside surface of each of the first, second, third, and fourth wall segments, herein “inner surfaces”) may have properties that minimize loss of liquid and reagent. These properties may be ubiquitous to the construction material itself or may be imparted upon one or more inner surfaces alone. Such properties include, but are not limited to, hydrophobicity, hydrophilicity, low permeability, resistance to binding of biochemical molecules (e.g., proteins, peptides, DNA, RNA, and the like), resistance to leaching, resistance to oxidation, resistance to reduction, low surface area, chemical stability (e.g., low reactivity), resistance to irradiation, and resistance to physical force (such as resistance to etching).

In any embodiment, one or more inner surfaces may exhibit properties that differ from the bulk material of the liquid reservoir, for example, through post-manufacture modification (e.g., physical or chemical modification) or the properties may be imparted in situ during manufacturing. For example, one or more inner surfaces may be treated or coated with a biologically inert material. For example, in any embodiment, one or more inner surfaces, e.g., of a PVDF-based liquid reservoir, may be coated or treated with a copolymer formed by zwitterionization of poly(styrene-r-4-vinylpyridine), zP(S-r-4VP). Other biologically inert coatings include, but are not limited to, silicon coatings, such as SILCONERT® (available from SilcoTek, Bellefonte, PA, USA), a carboxysilicon, such as DURSAN® coatings (also available from SilcoTek). In another example, one or more inner surfaces may be conjugated with antibodies for positive and negative selection-based sample preparation or with nucleic acids to serve as aptameric binding ligands or Watson-Crick base-pairing sequence specific binding ligand. In yet another example, one or more inner surfaces may be treated with a silane as a functional coating or with reagents suitable for use in Click Chemistry. In yet another example, one or more inner surfaces may be plasma treated for modification of water contact angle.

In yet another example, one or more of the inner surfaces may be polished to reduce the surface area arising from microporosity of the construction material. A high level of surface polish creates a surface that facilitates liquid beading and migration of any liquid beads to the low collection point of the reservoir, thereby minimizing reagent loss. Alternatively, one or more of the inner surfaces may be treated to impart a rough and therefore a higher surface area. Such treatment may be advantageous, for example, if the use of the liquid reservoir involves ligation of, e.g., a capture antibody.

Alternatively, or additionally, a liquid reservoir may be manufactured using surface modifying additives (SMAs), surface modifying macromolecules (SMMs), and/or surface modifying end groups (SMEs) to impart particularly desired properties to one or more inner surfaces of a liquid reservoir.

Methods of Manufacture

The methods by which a liquid reservoir as described and disclosed herein may be manufactured are not particularly limited and generally may be constructed by processes commonly used in polymer manufacturing. For example, in any embodiment, a liquid reservoir, as described herein, may be made by additive fusion deposition molding (FDM), additive selective laser sintering (SLS), additive stereolithography (SLA), reductive manual machining, reductive computer numerically controlled (CNC) machining, injection molding, blow molding, and vacuum forming.

As discussed above, it may be desirable to impart one or more properties to one or more inner surfaces of a liquid reservoir that differ from the properties of the bulk construction material, which may be accomplished in situ during manufacturing through the use of various additives or post-manufacturing by modifying one or more inner surfaces of a liquid reservoir.

The type of post-manufacturing surface modifications that may be implemented are not particularly limited and are well known to those of skill in the art. For example, one or more inner surfaces of a liquid reservoir may be subject to plasma discharge to oxidize the surface of the polymer, leaving underlying bulk layers unchanged. Such a treatment may change the contact angle of the polymer, e.g., create a more hydrophilic surface. In another example, functional molecules may be immobilized (e.g., conjugated) to one or more inner surfaces of the liquid reservoir. Such functional molecules include, but are not limited to, nucleic acids (e.g., RNAs, DNA), peptides, proteins (e.g., heparin, hirudin, albumin), antibodies, and the like. Other exemplary processes include, but are not limited to, ultraviolet irradiation, ion implantation, polishing, impregnation, etching, grafting, photo-lithography, or coating (e.g., a polymeric coating that differs from the primary construction material of the reservoir). One of skill in the art will be familiar with and be able to employ appropriate methods for such surface modifications.

Alternatively, or additionally, one or more surface modifying additives (SMAs), surface modifying macromolecules (SMMs), and/or surface modifying end groups (SMEs) may be incorporated during manufacturing to impart particularly desired properties to one or more surfaces of a liquid reservoir. SMMs are based on the use of an amphiphilic tri-block copolymer formed by a hydrophobic or hydrophilic segment, usually identical or compatible with the polymeric matrix, and end-capping block segments (silicones, fluorinated segments, olefins, and others) with low polarity, of which perfluorinated segments have been among the most commonly used. SMAs are amphiphilic di-block or tri-block copolymers where one of the blocks has higher affinity for the bulk material and the other block has little attraction for the base polymer, usually due to lower polarity or higher hydrophilicity. SMEs are not considered additives, but are part of the base polymer backbone itself.

Methods of Use

The liquid reservoirs may be used in any application where liquid retention is desired with additional advantages gained in automated applications where reagent recovery is important. Reagent recovery volume using any embodiment of a liquid reservoir as disclosed herein, particularly those with one or more low points, may be improved compared to other methods (e.g., pipette-based aspiration). Examples of reagents that may be collected in the liquid reservoirs described and disclosed herein are not particularly limited, but include, as non-limiting examples only, proteins, peptides, nucleic acids, nucleotides, spent cell culture media, prepared reagents, chemical intermediates, and the like.

For example, a liquid reservoir, as described herein may be used in next generation sequencing (NGS). After amplification by PCR, a well plate (typically a 384-well plate) can be inverted into a liquid reservoir as described herein and centrifuged to dispel all material from the well plate into the reservoir. Reagent can then be recovered from the liquid reservoir with little to no waste, particularly in embodiments with a single low point, for further processing. Advantageously, the liquid reservoirs may also be compatible with other laboratory equipment, such as the ClickBio® Bottomless Waste Station (available from ClickBio®, Reno, NV, USA) as well as other products available from ClickBio®.

In another example, any embodiment of a liquid reservoir as disclosed herein may be used for removing reagent and drying multi-well plates following chemical surface modification in a production environment.

EMBODIMENTS

What is Claimed is:

1. A reservoir for minimizing loss of a liquid, the reservoir comprising:a walled perimeter formed of at least one wall segment, the walled perimeter having a top edge and a bottom edge,and a floor portion joined to the walled perimeter, thereby forming a liquid space configured to hold at least one volume of liquid, the floor portion comprising at least one indentation and having a lowest point, wherein the at least one indentation is provided in the lowest point of the floor portion.
2. The reservoir of embodiment 1, wherein the walled perimeter comprises a first wall segment, a second wall segment, a third wall segment, and a fourth wall segment, wherein the first and third wall segments are parallel to each other and the second and fourth wall segments are parallel to each other.
3. The reservoir of embodiment 1, further comprising at least one partition wall oriented perpendicularly with one or more of a) the first and third wall segments and b) the second and fourth wall segments, thereby separating the liquid space into at least two subdivisions, each comprising a floor portion containing an indentation and having a lowest point, wherein the indentation is provided in the lowest point of each floor portion.
4. The reservoir of any one of embodiments 1-3, wherein the walled perimeter is rectangular-shaped.
5. The reservoir of any of embodiments 1-5, further comprising a projection on each of the first and third wall segments, the projection extending into the liquid space and positioned on the first and third wall segments at a nesting distance below the top edge of the walled perimeter.
6. The reservoir of any of embodiments 1-6, further comprising a projection on each of the second and fourth wall segments, the projection extending into the liquid space and positioned on the second and fourth wall segments at a nesting distance below the top edge of the walled perimeter.
7. The reservoir of any of embodiments 1-6, wherein the nesting distance is about 2 mm to about 2.5 mm.
8. The reservoir of any of embodiments 1-7, wherein the first wall segment and third wall segments each have an inside length of about 126.2 mm to about 127.3 mm and wherein the second wall segment and fourth wall segments each have an inside width of about 83.9 mm to about 85 mm.
9. The reservoir of any of embodiments 1-8, wherein each of the first wall segment, second wall segment, third wall segment, and fourth wall segment have a surface that face the liquid space and wherein the floor portion has at least one bottom surface facing the liquid space, wherein the four wall segment surfaces and the at least one bottom surface is resistant to one or more of the binding of protein, peptides, nucleotides, or nucleic acids.
10. The reservoir of any of embodiments 1-9, wherein the walled perimeter and the floor portion comprise a polymer.
11. The reservoir of any of embodiments 1-10, wherein the floor portion comprises an inverted cone or pyramid.
12. The reservoir of any of embodiments 1-11, wherein the floor portion comprises a rectangular pyramid.
13. The reservoir of any of embodiments 1-12, wherein the floor portion comprises an equilateral pyramid.
14. The reservoir of any of embodiments 1-13, wherein each of the first wall segment, second wall segment, third wall segment, and fourth wall segment has a thickness of about 0.55 mm to about 0.60 mm.
15. The reservoir of any of embodiments 1-14, wherein the top edge is rectangular-shaped.
16. The reservoir of any of embodiments 1-15, wherein the bottom edge is a rounded rectangle having four rounded corners.
17. The reservoir of any of embodiments 1-16, wherein each of the four rounded corners has a corner radius of about 1 mm to about 15 mm.
18. A reservoir for minimizing loss of a liquid, the reservoir comprising:a walled perimeter formed of at least one wall segment, the walled perimeter having a top edge and a bottom edge,and a floor portion joined to the walled perimeter, thereby forming a liquid space configured to hold at least one volume of liquid,wherein the reservoir has a length dimension and a width dimension, the floor portion contains at least one trough indentation spanning the length dimension and has a substantially two dimensional bottom, and the two dimensional bottom is provided in the lowest point of the floor portion.
19. The reservoir of embodiment 18, wherein the bottom edge is a rounded rectangle having four rounded corners.
20. The reservoir of embodiment 18 or 19, wherein the walled perimeter comprises a first wall segment, a second wall segment, a third wall segment, and a fourth wall segment, wherein the first and third wall segments are parallel to each other and the second and fourth wall segments are parallel to each other.
21. The reservoir of any of embodiments 18-20, wherein the walled perimeter is rectangular-shaped.
22. The reservoir of any of embodiments 18-21, further comprising a projection on each of the first and third wall segments, the projection extending into the liquid space and positioned on the first and third wall segment at a distance below the top edge of the walled perimeter.
23. The reservoir of any of embodiments 18-22, further comprising a projection on each of the second and fourth wall segments, the projection extending into the liquid space and positioned on the second and fourth wall segment at a nesting distance below the top edge of the walled perimeter.
24. The reservoir of any of embodiments 18-23, wherein the nesting distance is about 2 mm to about 2.5 mm.
25. The reservoir of any of embodiments 18-24, wherein the first and third wall segment each have an inside length of about 126.2 mm to about 127.3 mm and wherein the second and fourth wall segments each have an inside width of about 83.9 mm to about 85 mm.
26. The reservoir of any of embodiments 18-25, wherein each of the first wall segment, second wall segment, third wall segment, and fourth wall segment have a surface that face the liquid space and wherein the floor portion has at least one bottom surface facing the liquid space, wherein the four wall segment surfaces and the at least one bottom surface is resistant to binding with one or more of protein, peptides, nucleotides, or nucleic acids.