Liquid immersion microscope objective assembly and related systems and methods

Abstract

A liquid immersion microscope objective assembly includes an objective lens, a liquid delivery system for delivering immersion liquid to the lens of the objective lens, and a liquid return system for removing liquid from an area near the lens. The objective lens can be located below a sample container. Liquid is delivered to the lens so that a mass is formed between the lens and the sample container. Excess liquid is drained into the return system, which can be done via a notch or a V-shaped passage. In addition, the flow direction through the delivery system can be reversed so that liquid can also be removed from an area near the lens via the delivery system. The assembly can also be used to remove liquid from the bottom side of the sample container. The assembly can also include a leak sensor and / or an electrowetting device.

Description

[0001] Related Applications

[0002] This application claims priority to U.S. Patent Application No. 16 / 268,447, filed on February 5, 2019, the entire contents of which are incorporated herein by reference. Technical Field

[0003] The present invention relates to a liquid immersion microscope objective lens, in particular to a liquid immersion microscope objective lens for an automatic microscope. Background Art

[0004] Microscopy generally refers to a class of analytical techniques used to obtain magnified images of samples, which can be biological or non-biological materials. Light-based (optical) microscopy techniques include brightfield microscopy, darkfield microscopy, confocal microscopy, selective plane illumination microscopy (SPIM), and more recently, light-field microscopy. A typical light-based microscope includes: a sample stage on which is mounted the sample to be imaged; a light source for generating light for illuminating the sample; optics for directing the light as a beam to the sample and thereby illuminating the sample; an eyepiece and / or camera for taking an image of the sample; and optics for directing light emitted from the illuminated sample to the eyepiece and / or camera.

[0005] Microscope optics include an objective lens. An objective lens is typically a cylindrical structure encapsulated with one or more lenses, including a lens at one end (or end) of the objective lens closest to the sample. The primary function of the objective lens is to collect light emitted from the sample and focus the light to produce an amplified (e.g., 4 times, 10 times, 100 times, etc.) image that can be observed through an eyepiece and / or photographed by a camera. In some microscopes, the irradiated light from the light source can also pass through the objective lens to reach the sample. In this microscope, the objective lens is located below the sample, which can be referred to as an epifluorescence configuration.

[0006] In use, the lens at the end of the objective lens is positioned at a small distance from the sample. The gap between the objective lens and the sample can be an open space, that is, the gap can be an air gap. In this configuration, there are significant differences in the refractive index of various media along the optical path between the objective lens and the sample. Specifically, the refractive index of the lens, the air gap and the sample is different. In the case where the objective lens is located below or inverted (for example, in an epifluorescence configuration), the refractive index of the sample support (for example, a slide) on which the sample is mounted or the sample support (for example, a microplate) in which the sample is accommodated must also be considered. As a result, the sensitivity, resolution and image quality obtained by the microscope may be lower than expected. For example, due to excessive refraction at the gas-solid interface, some light from the sample will pass through the air gap without being collected by the objective lens, and therefore cannot be used for imaging signals.

[0007] To address this issue, microscopes can utilize a "liquid immersion" objective (or liquid immersion lens). This type of objective is constructed so that the lens at the end of the objective is immersed in an immersion medium (liquid immersion medium) (e.g., water or oil), which fills the gap between the lens and the sample support. The sample can also be immersed in the immersion medium. The immersion medium acts as a refractive index matching material. That is, the immersion medium has a higher refractive index than air, which is more closely matched to the refractive index of the lens, sample support, and sample. Therefore, the use of a liquid immersion objective can improve sensitivity (including at high magnifications), resolution, and image quality, at least in part by effectively increasing the numerical aperture (NA) of the objective lens.

[0008] However, compared to air-immersion objectives, liquid immersion objectives generally require more manual interaction to ensure that they function well in the intended manner in a microscope. Specifically, a large amount of manual interaction is required to successfully operate a liquid immersion objective and to use and handle the immersion medium. Due to the increased complexity and manual requirements, liquid immersion objectives have not generally been deployed in automated microscopes or high-volume screening systems. In automated or high-throughput operating environments, the workflow for manually operating liquid immersion objectives often fails. For example, as the speed requirements of operation increase, the manual labor required can take too long and / or have too high a rework / failure rate to be acceptable.

[0009] It would therefore be desirable to provide an improved liquid immersion microscope objective, particularly one suitable for use in automated or high volume / high throughput microscopy. Summary of the invention

[0010] In order to solve the aforementioned problems in whole or in part and / or problems that may have been observed by those skilled in the art, the present invention provides methods, processes, systems, devices, apparatuses and / or apparatuses as illustrated by way of example in the embodiments listed below.

[0011] According to an embodiment, an objective lens assembly includes: an objective lens, which includes a shell and a lens mounted at the shell, the lens including a lens outer surface; and a liquid handling device, which is mounted at the objective lens and is configured to immerse the lens outer surface in a liquid, the liquid handling device including: an edge, which surrounds the lens outer surface and defines a device opening for exposing the lens, the edge including a recess; a tapered portion, which includes a tapered surface connected to the recess; a delivery outlet, which is connected to the device opening; and a return inlet, which is connected to the tapered surface, wherein: the liquid handling device defines a liquid delivery path from the delivery outlet to the device opening; and the liquid handling device defines a liquid return path from the device opening, through the recess, along the tapered surface, and to the return inlet.

[0012] According to another embodiment, a method of immersing a lens of a microscope objective with liquid includes: positioning the microscope objective below a sample support so that the lens and the sample support are separated by a gap, wherein the lens is surrounded by a rim that defines an opening through which the lens is exposed, and the rim includes a recess; flowing liquid to the lens to form and maintain a liquid blob in the gap and to contact the lens and the sample support; and removing liquid while maintaining the liquid blob by flowing the liquid from the lens, into the recess, and along a conical surface located above the microscope objective.

[0013] According to another embodiment, a liquid immersion microscope objective lens assembly includes: an objective lens, which includes a shell and a lens mounted at the shell, the lens including a lens outer surface; and a liquid handling device, which is mounted at the objective lens and is configured to immerse the lens outer surface in liquid, the liquid handling device including: a rim, which surrounds the lens outer surface and defines a device opening that exposes the lens; a tapered portion, which includes a tapered surface; a delivery outlet, which is connected to the device opening; a return inlet, which is connected to the tapered surface; and an annular V-shaped passage, which is connected to the return inlet, wherein: the liquid handling device defines a liquid delivery path from the delivery outlet to the device opening; and the liquid handling device defines a liquid return path from the device opening, along the tapered surface, through the annular V-shaped passage, and to the return inlet.

[0014] According to another embodiment, a liquid immersion microscope objective lens assembly includes: an objective lens, which includes a shell and a lens mounted at the shell, the lens including a lens outer surface; and a liquid handling device, which is mounted at the objective lens and is configured to immerse the lens outer surface in liquid, the liquid handling device including: a liquid delivery system, which includes a delivery inlet, a delivery channel, and a delivery outlet openly connected to the lens, wherein the liquid delivery system defines a liquid delivery path from the delivery inlet, through the delivery channel, through the delivery outlet, and to the lens; and a liquid return system, which includes an annular passage, a return inlet, a return channel, and a return outlet, the annular passage being openly connected to the lens, wherein the annular passage has a V-shaped profile coaxial with the longitudinal axis, and the liquid return system defines a liquid return path from the lens, through the annular passage, through the return inlet, through the return channel, and to the return outlet.

[0015] According to another embodiment, a method of immersing a lens of a microscope objective with liquid includes: positioning the lens below a sample support so that the lens and the sample support are separated by a gap; flowing liquid to the lens to form and maintain a liquid blob in the gap and bring the liquid blob into contact with the lens and the sample support, and flowing liquid from the lens into an annular V-shaped passage surrounding the lens while maintaining the liquid blob.

[0016] According to another embodiment, a liquid immersion microscope objective system includes: a liquid immersion microscope objective assembly according to any of the embodiments disclosed herein; a liquid delivery line connected to a delivery inlet of the objective assembly; a liquid pump configured to establish a liquid flow from the liquid delivery line to a liquid delivery path of the objective assembly; and a liquid return line connected to a return outlet of the objective assembly.

[0017] According to another embodiment, the liquid immersion microscope objective system comprises a vacuum pump configured to establish a liquid flow from the liquid return path to the liquid return line.

[0018] According to another embodiment, the liquid immersion microscope objective system includes an aspiration line and a valve, wherein the valve is connected to the liquid delivery line and the aspiration line, and the valve is configured to switch between a first position and a second position, wherein: at the first position, the liquid in the liquid delivery line flows through the liquid delivery path to reach the delivery outlet; and at the second position, the liquid outside the liquid handling device flows into the delivery outlet, passes through the liquid delivery path, passes through the delivery inlet, and flows to the aspiration line.

[0019] According to another embodiment, the suction line is in communication with the liquid return line and, in the second position, liquid from outside the liquid treatment device flows through the suction line to the liquid return line.

[0020] According to another embodiment, a liquid immersion microscope objective system includes a vacuum pump configured to establish liquid flow from a liquid return path of the objective assembly to a liquid return line and liquid flow from a liquid delivery path to a suction line.

[0021] When viewing the following drawings and detailed description, those skilled in the art will or will become more easily understand other devices, equipment, systems, methods, features and advantages of the present invention. It is desirable that all such additional systems, methods, features and advantages are included in this description, within the scope of the present invention and protected by the appended claims. BRIEF DESCRIPTION OF THE DRAWINGS

[0022] The present invention can be better understood by referring to the following drawings. The components in the drawings are not necessarily drawn to scale, but are mainly used to illustrate the principles of the present invention. In the drawings, the same reference numerals throughout the different views represent corresponding parts.

[0023] Figure 1 is a schematic cross-sectional view of an example of a liquid immersion microscope objective assembly according to an embodiment of the present disclosure.

[0024] Figure 2 is a schematic diagram of an example of a liquid immersion microscope objective system according to an embodiment of the present disclosure.

[0025] Figure 3 is a perspective view of an example of a liquid immersion microscope objective lens assembly according to another embodiment of the present disclosure.

[0026] Figure 4 yes Figure 3 A cut-away elevation view of the liquid immersion microscope objective assembly shown in FIG.

[0027] Figure 5 yes Figure 3 A top view of the liquid immersion microscope objective assembly is shown in FIG.

[0028] Figure 6 is a schematic cross-sectional elevation view of a liquid immersion microscope objective lens assembly according to another embodiment of the present disclosure.

[0029] Figure 7 yes Figure 6 A top view of the liquid immersion microscope objective assembly is shown in FIG.

[0030] Figure 8 is a schematic cross-sectional elevation view of a liquid immersion microscope objective lens assembly according to another embodiment of the present disclosure.

[0031] Fig. 9 yes Figure 8 A top view of the microscope objective assembly shown in FIG.

[0032] Fig.10 is a schematic cross-sectional elevation view of a liquid immersion microscope objective lens assembly according to another embodiment of the present disclosure.

[0033] Fig.11 yes Fig.10 A top view of the microscope objective assembly shown in FIG.

[0034] Fig.12 is a perspective view of a liquid immersion microscope objective lens assembly according to another embodiment of the present disclosure.

[0035] Fig.13 yes Fig.12 A cut-away front view of the microscope objective assembly shown in FIG.

[0036] Fig.14 yes Fig.12 A perspective view of the pyramidal underside of the objective lens assembly is shown. DETAILED DESCRIPTION

[0037] As used herein, the term "sample" generally refers to one or more objects (or particles) that can be imaged by a microscope. For example, a sample can be a biological material (e.g., a biological cell, a multicellular tissue, a unicellular organism, a multicellular organism, etc.). Alternatively, a sample can be a non-biological material (e.g., a non-biological particulate matter).

[0038] For imaging by a microscope, a sample can be supported on or in a sample support at a specified position relative to the optics of the microscope, particularly the microscope objective. The sample is typically immersed in a liquid (e.g., a solution) on or in the sample support. The liquid can be composed as needed to perform functions related to, for example, sample preparation, sample processing, sample analysis, sample assay, etc. For example, the liquid can be or include a buffer, a preservative, a reagent, an imaging marker, etc.

[0039] As used herein, the term "sample support" (or "sample holder") generally refers to any structure used in a microscope to support a sample in a fixed position during imaging, although the position may be adjustable or movable if desired. For example, the sample support may be a solid substrate or plate (e.g., a microscope slide, a cover slip, etc.) on which the sample is supported, or a sample container (e.g., a well, a vial, a tube, a tray, etc.) in which the sample is supported or held. The sample support may be light-transmissive. In the case of a sample container, at least the bottom of the sample container may be light-transmissive if the image is to be taken from below the sample container. The sample container may be cylindrical or polygonal. The bottom of the sample container may be flat or tapered (conical or curved).

[0040] Specifically, in the case of sample containers, the "sample support" may include a group of such sample containers. Different sample containers may hold different samples, or may hold the same or similar samples but with different solutions, treatments or chemical compositions (e.g., different reagents). Multiple sample containers may be arranged in a one-dimensional (linear) or two-dimensional array. In a specific example, multiple sample containers may be arranged as a hole integrally formed as a part of a multiwell plate (also referred to as a microtiter plate, or "microplate", or optical plate). Typically, the holes of a microwell plate are arranged in a two-dimensional array with a row and column ratio of 2:3. Such a multiwell plate may have a standard form, such as a form with 24 holes, 96 holes, or 384 holes, or 1536 holes. Alternatively, multiple sample containers may be removed from the container support (e.g., vials removable from a rack).

[0041] As used herein, the term "light" generally refers to electromagnetic radiation that can be quantified as photons. As light relates to the present disclosure, light can propagate at wavelengths ranging from ultraviolet (UV) to infrared (IR). In the present disclosure, the term "light" is not intended to be limited to electromagnetic radiation in the visible light range. In the present disclosure, the terms "light", "photons" and "radiation" are used interchangeably.

[0042] Various embodiments disclosed herein relate to acquiring an image (or "imaging") of a sample. Depending on the embodiment and / or the sample being imaged, image acquisition can be based on fluorescence, reflection (or scattering), or transmission. Typically, imaging of a sample includes irradiating the sample with "excitation light" and collecting "emission light" emitted from the sample in response to the irradiation. In fluorescence-based imaging, the wavelength of the excitation light (e.g., in the ultraviolet (UV) range) is typically shorter (e.g., bluer) than (e.g., redder) the wavelength of the emission light. The source of fluorescence-based emission light can be autofluorescence from an object, or fluorescence from a fluorophore previously attached to the object. In reflection-based imaging, the wavelength of the excitation light can be approximately the same as the wavelength of the emission light. In this case, the light reflected (or scattered) from the object in response to the incident excitation light is the emission light. In transmission-based imaging, the excitation light incident on the object passes through the object and is attenuated due to absorption and / or scattering. In this case, the attenuated light radiated from the object is the emission light. In all of these cases, for convenience, in this disclosure, "excitation light" refers to the light used to illuminate the object, and "emission light" refers to the light collected from the sample in response to the illumination, regardless of whether the specific instance of acquiring the image is based on fluorescence, reflection, or transmission, unless otherwise stated or the context dictates otherwise.

[0043] As used herein, the term "fluid pipeline" or "liquid pipeline" generally refers to any type of conduit or multiple conduits used to guide a fluid (e.g., a liquid) from one point to another in a fluid system in a desired direction. Therefore, a "pipeline" can be or include one or more tubes (or pipes, etc.), passages, chambers, etc. The pipeline can also include other types of fluid components that perform functions other than conducting fluids. Other types of fluid components include, but are not limited to, pumps, valves, fluid accessories, fluid couplers, mixers, fluid flow mergers, heaters, coolers, pressure regulators, etc., as well as measuring instruments (e.g., temperature sensors, pressure sensors, flow sensors, etc.). A "pipeline" can include any combination of one or more tubes (or the like) and other types of fluid components.

[0044] Figure 1 1 is a schematic cross-sectional elevation view of an example of a liquid immersion microscope objective lens assembly (or device) 100 according to an embodiment of the present disclosure. For ease of illustration, Figure 1 100 and other figures include a Cartesian (XYZ) reference system, the origin of which is arbitrarily located relative to the objective lens assembly 100. The Z axis generally corresponds to the vertical direction. The objective lens assembly 100 as a structure can be considered to have a longitudinal axis L, and the length of the objective lens assembly 100 is defined along the longitudinal axis L. In this example, the longitudinal axis L corresponds to (coincides with or is parallel to) the Z axis.

[0045] The objective lens assembly 100 generally includes an objective lens 104 and a liquid handling device 108. The objective lens 104 includes an objective lens housing 112 and a primary entrance lens 116. The housing 112 extends from a first (upper) end (or the end of the objective lens 104) to a second (lower) end along a longitudinal axis L, with a first (upper) housing opening 120 located at the first end and a second (lower) housing opening 124 located at the second end. The lens 116 is mounted in the first housing opening 120 so that the outer surface of the lens 116 faces an area outside the housing 112. The housing 112 is generally cylindrical, with the longitudinal axis L corresponding to the central axis of the objective lens 104. A portion of the housing 112 is hollow and defines an optical path (generally along the longitudinal axis L) between the first housing opening 120 and the second housing opening 124. In addition to the lens 116 shown, as will be appreciated by those skilled in the art, the objective lens 104 may include other internal optical devices (not shown) that define the optical path, such as other lenses, reflectors, etc.

[0046] Figure 1 Also shown is a portion of a sample support that includes or supports a plurality of sample containers. In the example shown, the sample support is a microplate 128, and the sample container is an aperture 132 of the microplate 128. In use, one or more apertures 132 respectively accommodate one or more samples S to be imaged by an associated microscope. In the example shown, the objective lens assembly 100 is positioned below the microplate 128, which is sometimes referred to as an "inverted" configuration. In this case, at least the bottom 136 of the microplate 128 (or at least the portion of the bottom 136 below the aperture 132) is light-transmissive to allow an optical path through the bottom 136 to reach a selected sample S optically aligned with the lens 116 or from the selected sample S through the bottom 136. Therefore, when necessary, this configuration enables the target sample S to be illuminated (as shown by the excitation light 140) and imaged (as shown by the emission light 144) from below the microplate 128.

[0047] Thus, in the illustrated embodiment, the microscope including the illustrated objective 104 has an epi-fluorescence type configuration. In this case, the objective 104 receives (via the second housing opening 124) excitation light 140 at an excitation wavelength from an appropriate light source (e.g., a light emitting diode, a laser, etc.) via appropriate optical devices (e.g., an excitation filter, a dichroic beam splitter, a filter cube, a lens, a mirror, a light guide, an aperture, a confocal disk, etc.), and transmits the excitation light 140 to the sample S, thereby illuminating the sample S and inducing a fluorescence response of the sample S. The objective 104 also collects the resulting emission light 144 emitted from the sample S at an emission wavelength (different from the excitation wavelength in fluorescence applications), whereby the emission light 144 passes through the second housing opening 124 and is transmitted to an appropriate imaging device (e.g., a camera) via appropriate optical devices (e.g., an emission filter, a dichroic beam splitter, a filter cube, a lens, a mirror, a light guide, an aperture, a confocal disk, etc.). Thus, in this example, objective 104 focuses excitation light 140 onto a focal plane in the thickness of sample S, and focuses emission light 144 onto a sensing plane of the imaging device. In an epi-fluorescence configuration, a portion of the excitation light path and a portion of the emission light path may be defined by the same optical component, such as objective 104 as shown.

[0048] In an alternative embodiment, the objective lens 104 may be used only to collect the emission light 144 , and the excitation light 140 may be directed to the sample S from above the sample S.

[0049] As will be appreciated by those skilled in the art, the objective lens assembly 100 may be mounted to an objective stage (not shown). As indicated by arrow 148, the objective stage is configured to move (translate) the objective lens 104 along the Z-axis toward and away from the microwell plate 128 (and thus toward and away from the selected sample S). For example, the objective stage may be driven by a reversible stepper motor via a suitable drive train (e.g., a screw, a worm gear, etc.). Movement or adjustment of the objective lens assembly 100 along the Z-axis allows the objective lens 104 to focus on an image of the sample S and scan the thickness of the sample S along the Z-axis. The objective stage may also be movable relative to the microwell plate 128 along the X-axis and the Y-axis (usually in a horizontal plane) so that the objective lens 104 can be positioned to be optically aligned with the selected sample S for illumination and imaging.

[0050] The objective stage can also be movable (e.g., rotatable or linearly slidable) to enable selection of different objective lens assemblies 100 and switching of different objective lens assemblies 100 into the excitation light path and the emission light path. Different objective lens assemblies 100 can accommodate different objective lenses 104, for example, objective lenses 104 with different magnifications. Each such objective lens assembly 100 can be constructed as described herein. The construction and operation of the automatic objective stage are generally known to those skilled in the art and therefore do not need to be described in further detail in this disclosure.

[0051] As will be appreciated by those skilled in the art, the microplate 128 is typically supported by a sample stage system (not shown) and movable by the sample stage system. The sample stage system may include a sample stage on which the microplate 128 is mounted. The sample stage has an opening below the hole 132 that enables any hole 132 to be selectively found (addressed) by the objective lens 104. The sample stage system may also include one or more motorized slides mechanically connected to the sample stage. The sample stage system may include a motor (e.g., a reversible stepper motor) configured to drive the slide via a suitable drive chain (e.g., a screw, a belt, etc.). With this configuration, the slide moves (translates) the sample stage along the X-axis and Y-axis (typically in a horizontal plane), and thereby moves (translates) the microplate 128 and the sample S supported by the microplate 128, as shown in FIG. Figure 1 As shown by arrow 152 in FIG. 1 (in the case of X-axis movement). Figure 1 As shown, the movement of the microplate 128 relative to the objective 104 thus enables the selected sample S to be positioned to be optically aligned with the field of view of the objective 104 for illumination and imaging. Subsequently, the microplate 128 can be moved along the X-axis and / or Y-axis as needed to select another sample S contained in another well 132 and optically align the other sample S with the lens 116 of the objective 104. In some embodiments, the sample stage system can be additionally configured to move the microplate 128 along the Z-axis. The construction and operation of the automated sample stage system are generally known to those skilled in the art and therefore do not need to be described in further detail in the present disclosure.

[0052] Figure 1 The objective lens assembly 100 is shown in an operating position relative to the microwell plate 128 in which a selected sample S can be imaged. In the operating position, the lens 116 (the vertex) at the end of the objective lens 104 is separated from the bottom surface 160 of the microwell plate 128 by an axial gap 156, i.e., a distance along the Z axis. In a non-exclusive example, the size of the axial gap 156 (the distance in the Z direction) is in the range of from _ to _.

[0053] The liquid handling device 108 is configured to generate and maintain a liquid mass 164 on the lens 116. Specifically, the liquid handling device 108 is configured to generate and maintain a liquid mass 164 in the axial gap 156 so that the liquid mass 164 is in contact with both the lens 116 and the bottom surface 160 of the microplate. As described above, immersing the lens 116 in the liquid mass 164 increases the numerical aperture (NA) of the lens 116 and, therefore, increases the resolution (or resolving power) of the associated microscope. This is because the liquid or immersion medium of the liquid mass 164 has a higher refractive index than air. In a typical embodiment envisioned by the present subject matter, the immersion medium is water. Alternatively, the immersion medium can be a transparent oil, ethylene glycol, or other transparent liquid suitable for microscopy.

[0054] To generate and maintain the liquid bolus 164, the liquid handling device 108 includes a liquid delivery system and a liquid return system. The liquid delivery system and the liquid return system may be implemented by one or more structures 168 coaxially surrounding the objective 104. The liquid handling device 108 may be attached or mounted to the objective 104 in any suitable manner such that the liquid handling device 108 and the objective 104 are movable together as a unit.

[0055] The liquid delivery system may include a delivery inlet 172, a delivery channel 176, and a delivery outlet 180 in open communication with the lens 116. The components of the liquid delivery system are configured to define a liquid delivery path partially depicted by arrow 184 from the delivery inlet 172, through the delivery channel 176, through the delivery outlet 180, and to the lens 116 (i.e., to the area of ​​the lens 116 facing the outside of the objective 104 and on the outer surface facing the bottom side of the microwell plate 128). When the objective lens assembly 100 is installed in a microscope, the delivery inlet 172 is connected to a suitable liquid source 188 (e.g., a bottle or other type of container for supplying immersion medium), such as via tubing and fluid fittings.

[0056] The delivery channel 176 may represent one or more conduits (e.g., channels, cavities, etc.) that interconnect the delivery inlet 172 and the delivery outlet 180 as needed. As shown in the figure, at least a portion of the delivery channel 176 (where the arrow 184 is located) may be an annular conical space surrounding the upper portion (or end) of the objective lens 104. In one embodiment, as shown in FIG. Figure 1 As shown, the tapered space can be defined between the outer surface of the tapered portion of the objective housing 112 and the inner surface of the hollow tapered portion 192 of the structure 168 of the liquid handling device 108. Alternatively, the annular space can be formed as one or more channels completely inside the tapered portion 192 of the structure 168. In the embodiment shown, the delivery outlet 180 can be an annular opening around the lens 116. The annular configuration of the delivery outlet 180 and the adjacent tapered portion of the delivery channel 176 can help to evenly distribute the liquid on the lens 116 and mitigate the introduction of bubbles that may interfere with sample imaging.

[0057] In another embodiment, the delivery outlet 180 may be a single port positioned adjacent to the lens, and / or the delivery passage 176 may be a single passage (e.g., formed in the structure 168 of the liquid handling device 108) leading from the delivery inlet 172 to the single port of the delivery outlet 180. In another embodiment, the delivery outlet 180 may be a plurality of separate ports arranged around the lens 116 (i.e., circumferentially spaced from each other at different angular positions relative to the longitudinal axis L passing through the center of the lens 116). In this case, the delivery passage 176 may be a plurality of separate passages leading from the delivery inlet 172 to the respective ports of the delivery outlet 180.

[0058] The liquid return system may include a return inlet 106, a return channel 110, and a return outlet 114. The components of the liquid return system are configured to define a liquid return path, partially depicted by arrow 118, from the lens 116 along the outer surface of the tapered portion 192 of the structure 168, through the return inlet 106, through the return channel 110, and to the return outlet 114. When the objective lens assembly 100 is installed in a microscope, the return outlet 114 communicates with a suitable liquid receptacle 118 (e.g., a bottle or other type of container for recovering immersion medium), such as via tubing and fluid fittings.

[0059] The return inlet 106 may represent one or more openings that provide fluid communication between the outer surface of the tapered portion 192 of the liquid handling device 108 and the rest of the liquid return system. The return channel 110 may represent one or more conduits (e.g., channels, cavities, etc.) that fluidly interconnect the return inlet 106 and the return outlet 114 as desired.

[0060] In one embodiment, Figure 1 As shown, the liquid return system may also include an annular V-shaped (or triangular) passage 196, which is in open communication with the lens 116 via the outer surface of the tapered portion 192 of the liquid handling device 108. In this case, the V-shaped passage 196 is part of the liquid return path. In other words, the liquid return path extends along the outer surface of the tapered portion 192 into the V-shaped passage 196 and extends into the return inlet 106. The annular V-shaped passage 196 may be part of one or more outer surfaces of the structure 168 or formed on one or more outer surfaces of the structure 168. The annular V-shaped passage 196 has a V-shaped (or triangular) profile coaxial with the longitudinal axis L. On any elevational plane projected radially outward from the longitudinal axis L (for example, on a plane with respect to the longitudinal axis L), the annular V-shaped passage 196 may be formed on the outer surface of the tapered portion 192. Figure 1The V-shaped profile is obvious in both the XZ plane corresponding to the front view of the lens 116. The annular V-shaped passage 196 is located at a greater radial distance from the longitudinal axis L than the annular delivery outlet 180 surrounding the lens 116 (radial distance to the longitudinal axis L). In a non-exclusive example, the angle between the two surfaces of the V-shape forming the annular V-shaped passage 196 is in the range of 60 to 100 degrees. In this embodiment, the return inlet 106 can correspond to (or communicate with) the open annular lower vertex of the V-shaped passage 196.

[0061] In operation, the objective lens assembly 100 and the microplate 128 are moved to corresponding operating positions, such as Figure 1 As shown, in this operating position, the lens 116 is optically aligned with the selected sample S and positioned to have an axial gap 156 of a desired distance. A liquid (immersion medium) flow is established from a liquid source 188, through a liquid delivery system of the liquid handling device 108, and to the lens 116. A suitable fluid pump (not shown) in communication with the liquid source 188 may be provided to establish a liquid flow on the liquid delivery side of the liquid handling device 108. When the liquid is delivered to the lens 116 at a sufficient flow rate, a liquid mass 164 is formed in the axial gap 156 between the lens 116 and the bottom surface 160 of the microwell plate, and the liquid mass 164 contacts the lens 116 and the bottom surface 160 of the microwell plate, respectively. As a non-exclusive example, the flow rate of the liquid supplied to the objective lens assembly 100 to form the liquid mass 164 may be in the range of 1 mL / min to 40 mL / min. The formation, retention, size, and shape of liquid mass 164 may depend, at least in part, on the size of axial gap 156 and the surface tension of the liquid, in addition to the liquid flow rate through the liquid delivery system.

[0062] After completing the imaging of the selected sample S, the objective lens assembly 100 and / or the microwell plate 128 can be moved to another operating position as needed to image another sample S located in another hole 132 of the microwell plate 128. During this movement, the liquid can continue to circulate on the lens 116 between the liquid delivery system and the liquid return system as needed. Therefore, the lens 116 can remain wet even when the liquid mass 164 is not maintained. Keeping the lens 116 wet can help remove particles and prevent particles from accumulating on the lens 116. Such particles can interfere with sample imaging and damage the lens 116 or other optical devices of the microscope. Keeping the lens 116 wet can also be used to prime the lens 116 in advance to subsequently form a liquid mass 164. After repositioning the objective lens assembly 100 and / or the microwell plate 128 for imaging another sample S, the liquid mass 164 is then formed again. The liquid flow rate through the liquid delivery system can be adjusted as needed to alternate between forming the liquid bolus 164 and simply keeping the lens 116 wet (such as for cleaning the lens 116). For example, when the liquid bolus 164 is not being maintained, the liquid flow rate can be reduced or stopped.

[0063] When the liquid flow to the lens 116 at the desired liquid flow rate maintains the liquid mass 164, any excess liquid on or near the top of the objective lens assembly 100 (i.e., liquid that is not used to maintain the liquid mass 164 or to keep the lens 116 wet) drains downward along the outer surface of the tapered portion 192 and into the annular V-shaped passage 196. The excess liquid is then removed by the liquid return system of the liquid handling device 108 and collected at the liquid receiver 118. A suitable fluid pump (not shown) in communication with the liquid receiver 118 may be provided to facilitate liquid flow on the liquid return side of the liquid handling device 108. For example, the fluid pump may create suction (vacuum) at the junction of the bottom of the annular V-shaped passage 196 and the return inlet 106.

[0064] The annular V-shaped configuration of the passage 196 facilitates continuous circulation of liquid between the liquid delivery system and the liquid return system on the lens 116, and also facilitates rapid capture and removal of excess liquid from the end of the objective lens 104. The annular V-shaped configuration of the passage 196 also promotes coalescence of individual droplets of excess liquid into larger droplets or streams of liquid, thereby facilitating the establishment of liquid flow through the liquid delivery system. In an embodiment, the open V-shaped passage 196 works in conjunction with a hydrostatic pressure mechanism and delivers liquid to an adjacent horizontally configured surface tension passage 441, which will be discussed below in conjunction with Figure 4 Further description.

[0065] In an embodiment, one or both of the surfaces forming the annular V-shaped passage 196 may be treated to increase the hydrophilicity of the surface (i.e., increase the wettability of the surface, or reduce the hydrophobicity of the surface), thereby enhancing the ability of the liquid to drain into the annular V-shaped passage 196 and drain from the bottom of the annular V-shaped passage 196. For example, as understood by those skilled in the art, the surface of the annular V-shaped passage 196 may be treated by a suitable surface functionalization technique or by depositing a suitable coating (film, layer or material, etc.) to increase the hydrophilicity. The outer surface of the tapered portion 192 of the liquid handling device 108 may also be treated to increase the hydrophilicity of the outer surface.

[0066] In an embodiment, the liquid handling device 108 can be configured to induce an electrowetting effect on one or both of the surfaces forming the annular V-shaped passage 196 to increase the hydrophilicity of the surface. In such an embodiment, the surface of the annular V-shaped passage 196 is made of a conductive material (e.g., a metal, a metal alloy, or a conductive coated plastic, etc.), or can additionally be coated with a naturally hydrophobic electrically insulating material (e.g., a fluoropolymer such as polytetrafluoroethylene). The electrowetting effect can be induced by applying a voltage potential to one or both of the surfaces. As will be appreciated by those skilled in the art, the voltage potential renders the surface more hydrophilic by reducing the contact angle (and interfacial energy) between the liquid and the surface, causing the liquid to tend to spread out on the surface and flow more easily along the surface. The following is in conjunction with Figure 2 Another example of an electrowetting configuration is described.

[0067] like Figure 1 As shown in one example of a position in FIG. 1 , the excess liquid can include residual liquid 122 that adheres to the bottom surface 160 of the microplate 128 (e.g., due to a previously formed liquid mass 164). Such residual liquid 122 is undesirable because it may separate from the microplate 128 and drip onto sensitive mechanical, optical, or electronic components of the microscope, potentially causing contamination and / or damage. To prevent contact between the residual liquid 122 and these components, the residual liquid 122 can be captured and removed by the liquid handling device 108. To prevent contact between the residual liquid 122 and these components, the residual liquid 122 can be captured and removed by the annular V-shaped passage 196 when the residual liquid 122 drips onto the outer surface of the tapered portion 192 of the liquid handling device 108, or directly drips into the annular V-shaped passage 196, or is otherwise discharged into the annular V-shaped passage 196 by suction.

[0068] Alternatively or additionally, in embodiments, the liquid delivery system can be configured to remove excess liquid (including residual liquid 122) when not being used to deliver liquid to lens 116. Excess liquid is drained into delivery outlet 180 and removed via the liquid delivery system. In such embodiments, the liquid delivery system can be referred to as a liquid delivery / recovery system. Figure 2 Another example of such an embodiment is described.

[0069] Thus, the liquid delivery system and / or liquid return system can be used to capture excess liquid and clean and dry the bottom surface 160 of the microplate 128 and the end of the objective lens 104. Other techniques for cleaning and drying these components can be utilized, such as air flow, application of other liquids (such as alcohol, etc.), and application of drying and conditioning pads.

[0070] In addition, a hydrophobic wiper or roller can be passed across the top of the objective lens 112 and the liquid handling device 108 to transfer the liquid, thereby collecting the liquid into the annular V-shaped passage 196. The residual liquid 122 thereby drips onto the outer surface of the tapered portion 192 of the liquid handling device 108, or directly drips into the annular V-shaped passage 196, or is otherwise drained into the annular V-shaped passage 196 by surface tension and wetting. With the help of this wiper or roller, the liquid mass 164 can be reset and completely removed, or bubbles on the top surface of the objective lens 104 can be transferred.

[0071] Figure 2 2 is a schematic diagram of an example of a liquid immersion microscope objective lens system 200 according to an embodiment of the present disclosure. The system 200 includes an objective lens assembly 250 (in Figure 2 ), a liquid source 288 (e.g., a supply bottle), a liquid receiver 218 (e.g., a recovery bottle), and other fluid handling components (e.g., conduits, pumps, valves, etc.) required for the objective lens assembly 250 and associated system 200 to operate in the manner described herein.

[0072] Objective lens assembly 250 may be constructed according to any of the embodiments described herein, such as Figure 1 Thus, the objective lens assembly 250 includes an objective lens (e.g., as described above and in Figure 1 The objective lens 104 shown in FIG. 1 and the liquid handling device 208 attached to the objective lens (in FIG. Figure 2 In addition, the liquid handling device 208 includes a liquid delivery system and a liquid return system, the liquid delivery system is in fluid communication with the liquid source 288 via the delivery inlet 272, and the liquid return system is in fluid communication with the liquid receiver 218 via the return outlet 214.

[0073] In this embodiment, the liquid handling device 208 includes a hollow conical portion 292 that is located above and around the conical end of the objective lens and has an opening for accommodating the light path through the objective lens (e.g., Figure 1 292 and the annular cover 226. In the present embodiment, the conical portion 292 and the annular cover 226 are physically separated components that are spaced apart from each other (such as by a spacing member (e.g., a gasket)). The spacing member can be formed of an electrically insulating material. The liquid handling device 208 also includes an annular V-shaped (or triangular) passage 296 as described herein. The annular V-shaped passage 296 includes an inner surface 230 and an outer surface 234 that define a V-shaped (or triangular) profile of the passage 296. The inner surface 230 can be a portion of the conical portion 292, and the outer surface 234 can be a portion of the annular cover 226. In the case where the conical portion 292 and the annular cover 226 are separate components, the lower vertex of the annular V-shaped passage 296 can be an annular opening that communicates with a fluid component of the liquid return system (such as, the return outlet 214).

[0074] The liquid delivery system of the liquid handling device 208 is in fluid communication with the liquid source 288 via the liquid delivery line 238. The liquid pump 242 is operably positioned in communication with the liquid source 288, such as in the liquid delivery line 238 between the liquid handling device 208 and the liquid source 288, to establish a liquid flow to the liquid handling device 208 at a desired flow rate (or pressure). The liquid pump 242 can be any type suitable for pumping the immersion medium at the flow rate desired by the present subject matter. For example, various types of positive displacement pumps may be suitable, a non-exclusive example of which is a peristaltic pump.

[0075] The liquid return system of the liquid handling device 208 is in fluid communication with the liquid receiver 218 via a liquid return line 246. A liquid return pump 254 is operably positioned in communication with the liquid receiver 218 to establish a negative pressure differential in the liquid return line 246 between the annular V-shaped passage 296 and the liquid receiver 218, whereby liquid is drawn into the annular V-shaped passage 296 via suction (vacuum). In the illustrated embodiment, the liquid return pump 254 is a vacuum pump positioned downstream of the liquid receiver 218. Alternatively or additionally, a liquid pump similar to the liquid pump 242 described above may be positioned in the liquid return line 246 between the liquid handling device 208 and the liquid receiver 218.

[0076] The two-way valve 258 may be operably positioned in the liquid return line 246 between the liquid handling device 208 and the liquid receiver 218. The two-way valve 258 is configured to switch between a first (or open, or ON) position or switch state (as shown) that allows suction to be actively applied at the annular V-shaped passage 296 and a second (or closed, or OFF) position or switch state that shuts off suction (breaks the vacuum) in the liquid return line 246 and, therefore, shuts off suction at the annular V-shaped passage 296.

[0077] As described above, the liquid delivery system can be configured to remove liquid from the end of the objective lens assembly 250 and the area above the objective lens assembly 250 when it is not being used to deliver liquid to the objective lens assembly 250. To this end, the system 200 can be configured to reverse the pressure differential and thus reverse the direction of the liquid flow in the liquid delivery system so that the liquid is removed from the outlet of the liquid delivery system near the lens of the objective lens (e.g., Figure 1 The system 200 may further include a suction line 258 for this purpose. The suction line 258 may then be selectively connected to the portion of the liquid delivery line 238 that is connected to the delivery inlet 272. For example, the system 200 may include a three-way valve 262 that may be switched between a first position or switching state that fluidly couples the liquid handling device 208 (at the delivery inlet 272) to the liquid delivery line 238 and a second position or switching state that fluidly couples the liquid handling device 208 to the suction line 258. In the specifically illustrated embodiment, one side of the three-way valve 262 is connected to a common liquid line 266 that is in turn connected to the delivery inlet 272. Depending on the position of the three-way valve 262, the common liquid line 266 is alternately used as a delivery line to the liquid handling device 208 and a suction line from the liquid handling device 208. The other side of the three-way valve 262 is alternately connected to the liquid delivery line 238 and the suction line 258 according to its position.

[0078] In the first position of the three-way valve 262 (as shown), the liquid delivery line 238 is connected (active) and the suction line 258 is not connected (inactive). Therefore, under the action of the liquid pump 242, the first position allows liquid to be delivered from the liquid source 288 to the liquid treatment device 208 via the liquid delivery line 238, the three-way valve 262 and the common liquid line 266. In the second position, the liquid flow path through the three-way valve 262 is switched so that the suction line 258 is currently connected and the liquid delivery line 238 is currently not connected. Therefore, the second position allows liquid to be sucked from the liquid delivery system of the liquid treatment device 208 into the suction line 258 via the common liquid line 266 and the three-way valve 262. When the second position is on, the liquid pump 242 can be turned off. Additionally, the two-way valve 258 may be activated during aspiration to increase the flow rate through the common liquid line 266, thereby using the liquid return pump 254 to clean the lines 266, 258 or to draw liquid from the bottom surface 160 of the microplate 128 by suction at the delivery outlet 180. The two-way valve 258 may be activated while the liquid return pump 254 is operating to test the vacuum level created in the liquid receiver 218.

[0079] In this embodiment and as shown, the suction line 258 is coupled to the liquid return line 246 via any suitable fitting (e.g., a tee fitting) at a location upstream of the liquid receiver 218. This configuration allows both the suction line 258 and the liquid return line 246 to utilize the same liquid receiver 218 and vacuum source (liquid return pump 254). Alternatively, the suction line 258 may be coupled to a separate liquid receiver and vacuum source (not shown).

[0080] like Figure 2 As further shown in FIG. 2 , the system 200 may include a system controller 270, or the system 200 may communicate with a system controller 270. The system controller 270 may represent one or more electronically based modules (e.g., may include one or more electronically based processors, memories, other hardware, firmware, and / or software) configured to control the operation of the system 200, including the delivery of liquid to the objective lens assembly 250 and the removal of liquid from the objective lens assembly 250. Thus, as shown in FIG. Figure 2As partially depicted in the liquid delivery / aspiration side of the system 200 in FIG. 1 , the system controller 270 can communicate with the liquid pump 242, the three-way valve 262, the liquid return pump 254, and / or the two-way valve 258 via a wired or wireless communication link. With this configuration, the system controller 270 can control the liquid flow rate and flow path to and from the liquid handling device 208, including controlling the switching state of the three-way valve 262 and the two-way valve 258 as needed to wet the objective lens, create and maintain liquid boluses, and remove excess liquid as described herein.

[0081] In an embodiment, the system controller 270 may also represent a system controller of an associated microscope. Thus, the system controller 270 may be configured to control the operation of other components of the microscope, such as a light source, a camera, a stage for moving the objective lens assembly 250 and a sample support, a wavelength / filter selector, a data acquisition device, and an image processor, etc.

[0082] like Figure 2 As further shown in FIG. 2 , the system 200 may include a leak detector 274 configured to detect liquid leakage from the top of the liquid handling device 208. For example, the leak detector 274 may be configured to detect whether liquid has not been captured by the annular V-shaped passage 296, but has flowed radially outward beyond the annular V-shaped passage 296 and through the outer side(s) of the liquid handling device 208. In an embodiment, the leak detector 274 includes a leak sensor 278 located at any location on the liquid handling device 208 or at any location in the liquid immersion microscope objective system 200 where the presence of the inspection liquid is desired. For example, the leak sensor 278 may be located at a portion of the liquid handling device 208 that is located at a greater radial distance from the longitudinal axis of the liquid handling device 208 than the annular V-shaped passage 296. For example, the leak sensor 278 may be located at the periphery or outer side(s) of the liquid handling device 208. In an embodiment, the leak sensor 278 may include two or more electrodes, for example, attached to the periphery or the outside (s) of the liquid handling device 208. The leak sensor 278 may also include appropriate leak detection electronics configured to detect changes in electrical characteristics (e.g., conductance, capacitance, resistance) of the leak sensor 278 in the presence of liquid on and / or between the electrodes, thereby indicating the occurrence of a leak.

[0083] In an embodiment, the leak detection electronics may be part of the system controller 270, or the leak detection electronics may be in communication with the system controller 270. As a non-exclusive example of operation, the system controller 270 (or the leak detection electronics) receives feedback signals from the leak detector 270. Based on these signals, if it is determined that a leak has occurred or is occurring, the system controller 270 may send a control signal to the liquid pump 242, the control signal being adapted to cause the liquid pump 242 to shut down or reduce the flow rate of liquid being delivered to the liquid handling device 208. As another option, the system controller 270 may additionally send a control signal to the three-way valve 262, the control signal being adapted to cause the three-way valve 262 to switch from a first (liquid delivery) position to a second (liquid suction) position, thereby drawing liquid into the delivery inlet 272.

[0084] As yet another option, the system controller 270 may additionally send control signals suitable for adjusting the operation of the liquid return pump 254 and / or the two-way valve 258 to control the operation of the liquid return side of the system 200 in the event that a leak is detected. For example, the system controller 270 may begin applying a vacuum at the annular V-shaped passage 296 by switching the two-way valve 258 to the first (open or ON) position and activating the liquid return pump 254 (if not already activated). As another example, the system controller 270 may adjust the operation of the liquid return pump 254 to increase the level of vacuum applied in the event that a leak is detected.

[0085] like Figure 2 As further shown in FIG. 2 , the system 200 may include an electrowetting device 282 configured to induce an electrowetting effect in the annular V-shaped passage 296 and / or on other surfaces of the liquid treatment device 208. The electrowetting device 282 may include an electrowetting voltage source 286 electrically coupled to the inner surface 230 and / or the outer surface 234 of the annular V-shaped passage 296 (or to other surfaces electrically connected to the inner surface 230 and / or the outer surface 234) via electrical conductors, contacts, etc. The electrowetting voltage source 286 may be part of an electronic device that communicates with the system controller 270 (or is part of the system controller 270) to be able to control the operation of the electrowetting voltage source 286, including adjusting the voltage potential applied thereby.

[0086] As described above, the microscope can be provided with a plurality of objective lens assemblies 100 including different objective lenses 104, and one of the objective lens assemblies 100 can be selected at any given time to initiate operation. The above-described system 200 can be configured as needed to facilitate selection and switching out of different objective lens assemblies 100. For this purpose, additional fluid components (e.g., pumps, valves, pipes, etc.) can be appropriately provided and arranged, as will be appreciated by those skilled in the art.

[0087] Figures 3 to 5 An example of a liquid immersion microscope objective assembly 300 according to another embodiment of the present disclosure is shown. Figure 3 is a perspective view of the objective lens assembly 300. Figure 4 yes Figure 3 A cross-sectional front view of the objective lens assembly 300 is shown in FIG. Figure 5 yes Figure 3 The objective lens assembly 300 and its operation can be similar to the above-mentioned Figure 1 and Figure 2 The objective lens assemblies 100 and 250 shown respectively are similar. Thus, the objective lens assembly 300 generally comprises an objective lens 304 and a liquid handling device 308 arranged on the longitudinal axis L of the objective lens 304.

[0088] Objective 304 includes an objective housing 312 and a lens 316. The (generally cylindrical) housing 312 extends along a longitudinal axis L from a first (upper) end (or objective end) to a second (lower) end, with a first (upper) housing opening 320 located at the first end and a second (lower) housing opening 324 located at the second end. Lens 316 is mounted at or near the first housing opening 320 so that the outer surface of lens 316 faces away from objective housing 312 (i.e., facing the area outside and above housing 312). Thus, lens 316 is exposed through first housing opening 320. A portion of housing 312 is hollow, and defines an optical path (generally along longitudinal axis L) between first housing opening 320 and second housing opening 324, including through the illustrated lens 316 and other internal optical devices (not shown) provided by objective 304.

[0089] The liquid handling device 308 is mounted or positioned on the upper portion of the objective lens 304 and coaxially surrounds the upper portion of the objective lens 304. Figure 4 As shown, the liquid handling device 308 includes an annular seat 422 having a triangular or wedge-shaped profile. An O-ring 423 is disposed in the seat 422 and contacts the outer surface of the objective lens housing 312. The O-ring 423 in combination with the planar contact is used to fix the liquid handling device 308 and the objective lens 304 and keep the liquid handling device 308 and the objective lens 304 aligned.

[0090] The liquid handling device 308 includes a liquid delivery system and a liquid return system as described herein. Thus, the liquid handling device 308 is configured to perform the various functions described herein, including immersing the lens 316 (particularly the outer surface of the lens) in the immersion medium, and removing liquid from the lens 316 and the surrounding objective tip and nearby areas (such as the bottom of the sample support).

[0091] In this embodiment, the liquid handling device 308 includes an annular base 305, a hollow cone 309 and an annular cover 326 that cooperatively define a liquid delivery system and a liquid return system. The cone 309 includes a (first) cone 392 spaced above the end of the objective lens, and a flange portion 313 adjacent to the cone 392. The flange portion 313 is placed between the base 305 and the cover 326 and is separated from the base 305 and the cover 326 by corresponding annular spacing members 317 and 321. These components can be fixed together in a stacked arrangement by suitable fasteners such as threaded members (e.g., screws, bolts, etc.). The spacing members 317 and 321 can be made of any suitable elastomeric material, which can be used as a fluid seal or additionally as an electrical insulator. The cone 309 also includes an annular rim 325, which is coaxial with the longitudinal axis L and surrounds the lens 316. The rim 325 defines a device opening that exposes the lens 316. The edge 325 transitions to the tapered portion 392. In this document, the term "annular" encompasses not only a circular shape but also other arc shapes, such as an ellipse or an oval.

[0092] The liquid handling device 308 also includes a delivery inlet 372 and a return outlet 314 ( Figure 4 ), the delivery inlet 372 and the return outlet 314 can be implemented as fluid fittings that are coupled (e.g., by threads) to the internal passages in the base 305. The delivery inlet 372 and the return outlet 314 can be coupled to a pipe, which can be as described above and Figure 2 As shown in FIG. , it becomes part of the liquid pipeline.

[0093] The liquid delivery system includes a delivery inlet 372, a delivery channel, and a delivery outlet 380, wherein the delivery outlet 380 is in open communication with the lens 316 (i.e., the space above the outer surface of the lens in the device opening). The components of the liquid delivery system cooperatively define a liquid delivery path from the delivery inlet 372, through the delivery channel, through the delivery outlet 380, to the lens 316. The delivery channel can include one or more conduits (e.g., channels, cavities, etc.) as needed to interconnect the delivery inlet 372 and the delivery outlet 380. In this embodiment, as shown in FIG. Figure 4 As shown, the delivery channel includes an annular tapered space 376 defined between the outer surface of the (second) tapered portion 425 of the objective lens housing 312 and the inner surface of the tapered portion 392 of the cone 309. The tapered space 376 terminates at a delivery outlet 380. In this embodiment, the delivery outlet 380 is an annular opening defined between the edge 325 and the first housing opening 320, so that the delivery outlet 380 coaxially surrounds the first housing opening 320 and the lens 316. Figure 4As shown, the delivery channel can also include an internal conduit 429 formed in the cone 309. The internal conduit 429 extends from another conduit or cavity 433 (e.g., formed in the cone 309 and / or the base 305) that communicates with the delivery inlet 372 to an opening in the wall 437 of the cone 309, which in turn communicates with the tapered space 376.

[0094] In another embodiment, as an alternative to the annular conical space 376 and the annular delivery outlet 380, the delivery channel may include one or more conduits similar to the internal conduit 429 shown, but which leads from the delivery inlet 372 to one or more corresponding ports located at (or near) the edge 325 and thus opens into the space in the device opening above the lens 116.

[0095] As above combined Figure 2 As mentioned above, the liquid path defined by the liquid delivery system may also be used as a liquid suction (removal) path.

[0096] The liquid return system includes an annular V-shaped (or triangular) passage 396, a return inlet 306, a return channel and a return outlet 314 ( Figure 4 ), the annular V-shaped passage 396 is in open communication with the lens 316 (via the tapered portion 392). The components of the liquid return system cooperatively define a liquid return path from the lens 316, along the outer surface of the tapered portion 392, into the annular V-shaped passage 396, through the return inlet 306, through the return channel, and to the return outlet 314.

[0097] In this embodiment, the annular V-shaped passage 396 includes an annular inner surface 330 and an annular outer surface 334 that define the V-shaped (or triangular) profile of the V-shaped passage 396. As shown, the inner surface 330 can be a part of the tapered portion 392, and the outer surface 334 can be a part of the annular cover 326. At the lower vertex of the annular V-shaped passage 396, the inner surface 330 and the outer surface 334 are separated by an annular opening corresponding to the return inlet 306.

[0098] In an embodiment, the inner surface 330 and / or the outer surface 334 of the annular V-shaped passage 396 may be treated to increase hydrophilicity as described herein. Alternatively or additionally, one or more other surfaces of the liquid handling device 308 that may be exposed to the liquid may be treated in the same manner. Alternatively or additionally, the inner surface 330 and / or the outer surface 334 and / or one or more other surfaces of the liquid handling device 308 may be placed in communication with an electrowetting device as described herein.

[0099] The return channel of the liquid return system may include one or more conduits (e.g., channels, cavities, etc.) as desired to interconnect the return inlet 306 and the return outlet 314 fluidly. Figure 4 As shown, the return passage includes a horizontally configured annular surface tension passage 441 that communicates with the V-shaped passage 396 via the return inlet 306. The V-shaped passage 396, the return inlet 306, and the surface tension passage 441 are configured (i.e., in terms of size and shape) so that the V-shaped passage 396 works in conjunction with the mechanism of hydrostatic pressure and surface tension to transport liquid to the adjacent surface tension passage 441. When liquid fills the V-shaped passage 396, the accumulation of hydrostatic pressure forces the liquid into the surface tension passage 441. This in turn pulls the liquid into the surface tension passage 441 from anywhere around the 360 ​​degree perimeter of the V-shaped passage 396 via surface tension. In the example shown, the surface tension passage 441 is defined between the stacked portion of the annular cover 326 and the flange portion 313 of the cone 309.

[0100] Likewise Figure 4 As shown, the return passage can also include an internal conduit 445 formed in the flange portion 313 of the cone 309. The internal conduit 445 extends from another conduit or cavity 449 (e.g., formed in the cone 309 and / or the base 305) that communicates with the return outlet 314 to an opening in the wall of the surface tension passage 441 below the V-shaped passage 396.

[0101] like Figure 3 As best shown in FIG. 1 , objective lens assembly 300 also includes a leak sensor 378. Leak sensor 378 includes one or more leak sensing electrodes, such as electrodes 353 and 357, which are attached to (all or part of) the periphery or sidewall of substrate 305 and extend around (all or part of) the periphery or sidewall. Leak sensing electrodes 353 and 357 are in communication with respective electrical terminals or contacts 361 and 365, which may also be in communication with a suitable leak detector, such as those described above and Figure 2 Leak detector 274 is shown.

[0102] Figure 6 is a schematic cross-sectional elevation view of a liquid immersion microscope objective lens assembly 600 according to another embodiment of the present disclosure. Figure 7 yes Figure 6 A top view of the objective lens assembly 600 is shown in FIG.

[0103] As in other embodiments, the objective lens assembly 600 generally includes an objective lens 604 and a liquid handling device 608 disposed on the longitudinal axis of the objective lens 604. In addition, the objective lens 604 includes a lens 616 mounted at or near the housing opening of the objective lens 604, such that the lens 616 is exposed through the housing opening. As in other embodiments, the liquid handling device 608 is mounted or positioned at an upper portion of the objective lens 604 and coaxially surrounds the upper portion, as disclosed in other parts of the present disclosure. In addition, the liquid handling device 608 includes a liquid delivery system and a liquid return system as described herein. Therefore, the liquid handling device 608 is configured to perform various functions described herein, including immersing the lens 616 (particularly the outer surface of the lens) into the immersion medium, and removing liquid from the lens 616 and the surrounding objective lens end and nearby areas (such as, the bottom of the sample support).

[0104] Similar to Figures 3 to 5 In the illustrated embodiment, the structure of the liquid handling device 608 may include an annular base 605, a hollow cone 609, and an annular cover 626 that cooperatively define a liquid delivery system and a liquid return system. The cone 609 includes a tapered portion 692 spaced above the end of the objective lens. The cone 609 also includes an annular rim 625 that is coaxial with the longitudinal axis (through the center of the lens 616) and surrounds the lens 616. The rim 625 defines a device opening 627 that exposes the lens 616. The rim 625 transitions to the tapered portion 692. In this embodiment, the rim 625 has a discontinuity in its geometry, such as, Figure 7 Notch 629 is shown and will be described further below.

[0105] As in other embodiments, the liquid handling device 608 may generally include a delivery inlet 672 and a return outlet 614, which are implemented as fluid fittings that couple to the internal channels of the liquid handling device 608. In addition, the delivery inlet 672 and the return outlet 614 can be coupled to a fluid system, such as those described above and in Figure 2 A liquid immersion microscope objective system 200 is shown in FIG.

[0106] The liquid delivery system includes a delivery inlet 672, a delivery channel, and a delivery outlet 680 in open communication with the lens 616. The components of the liquid delivery system cooperatively define a liquid delivery path from the delivery inlet 672, through the delivery channel, through the delivery outlet 680, to the lens 616. The delivery channel can include one or more conduits (e.g., channels, cavities, etc.) as needed to interconnect the delivery inlet 672 and the delivery outlet 680. For example, as described above in conjunction with Figures 3 to 5In the embodiment shown, the delivery channel may include an annular conical space 676 between the delivery inlet 672 and the delivery outlet 680, and the delivery outlet 680 may be an annular opening, and the delivery channel may also include other internal conduits. Alternatively, as also described above, the delivery channel may include one or more conduits leading from the delivery inlet 672 to one or more corresponding ports located at or near the edge 625.

[0107] As described in other embodiments, as described above in conjunction with Figure 2 As mentioned above, the liquid path defined by the liquid delivery system may also be used as a liquid suction (removal) path.

[0108] The liquid return system includes a notch 629, a return inlet 606, and a return outlet 614, wherein the notch 629 is in open communication with the lens 616 (e.g., the space above the outer surface of the lens 616 surrounded by the rim 625). The components of the liquid return system cooperatively define a liquid return path from the lens 616 and the device opening 627, through the notch 629, along the tapered surface (e.g., the outer surface of the tapered portion 692), into the return inlet 606, and to the return outlet 614.

[0109] In some embodiments, the return inlet 606 and the return outlet 614 can be part of the same fluid component (eg, fitting). A return passage (eg, internal conduit) can be provided to fluidly interconnect the return inlet 606 and the return outlet 614 as desired.

[0110] The notch 629 may be formed on (as part of) the tapered portion 692. By presenting a geometric discontinuity near an area where a liquid mass is formed on the lens 614, the notch 629 captures liquid in that area and draws the liquid into the notch 629. In this manner, the notch 629 directs the liquid to flow along the outer surface of the tapered portion 692 on the side of the lens 614 where the notch 629 is located, and into the return inlet 606, which is located on the same side of the lens 614 as the notch 629. The liquid flows in Figure 6 and Figure 7 It is roughly indicated by arrows.

[0111] In order to collect the liquid entering the recess 629 and guide the liquid to flow back to the inlet 606, the liquid treatment device 608 may further include a liquid return passage (or overflow passage) 631 ( Figure 7), the liquid return passage 631 extends from the recess 629 toward the return inlet 606. In one embodiment, the return passage 631 is disposed on the tapered portion 692. That is, the return passage 631 can be positioned on the tapered portion 692, or formed on the tapered portion 692 (e.g., as part of the surface of the tapered portion 692). Generally, the return passage 631 has a length between the recess 629 and the return inlet 606, and a width transverse to the length. In one embodiment, as Figure 7 As shown, the width of the return passage 631 increases along the length of the return passage 631 in a direction away from the recess 629, that is, the return passage 631 gradually expands or fans outward from the recess 629. In addition, in order to facilitate the collection of liquid entering the recess 629, the liquid treatment device 608 may also include a concave basin (or bucket) 633 located between the return passage 631 and the return inlet 606, and the concave basin 633 is connected to the return passage 631 and the return inlet 606. For example, the concave basin 633 can be located (or formed in a part of the structure of the liquid treatment device 608) adjacent to the tapered portion 692. The size of the concave basin 633 can be significantly larger than the size of the recess 629, the return passage 631, and the return inlet 606. Therefore, in some embodiments, in addition to the recess 629 and the return inlet 606, the liquid return path includes the return passage 631, or includes both the return passage 631 and the concave basin 633.

[0112] Objective lens assembly 600 may otherwise be similar to that described above and as described above. Figures 3 to 5 The objective lens assembly 300 shown in FIG. For example, the objective lens assembly 600 may include the leakage sensor described herein. The objective lens assembly 600 may be used with a sensor such as the one described above and Figure 2 The system is used in combination with the liquid immersion microscope objective system 200 shown in FIG.

[0113] Figure 8 is a schematic cross-sectional elevation view of a liquid immersion microscope objective lens assembly 800 according to another embodiment of the present disclosure. Fig. 9 yes Figure 8 A top view of the objective lens assembly 800 is shown in FIG.

[0114] As in other embodiments, the objective lens assembly 800 generally includes an objective lens 804 and a liquid handling device 808 disposed on the longitudinal axis of the objective lens 804. In addition, the objective lens 804 includes a lens 816 mounted at or near the housing opening of the objective lens 804, such that the lens 816 is exposed through the housing opening. As in other embodiments, the liquid handling device 808 is mounted or positioned at an upper portion of the objective lens 804 and coaxially surrounds the upper portion, as disclosed in other parts of the present disclosure. In addition, the liquid handling device 808 includes a liquid delivery system and a liquid return system as described herein. Therefore, the liquid handling device 808 is configured to perform various functions described herein, including immersing the lens 816 (particularly the outer surface of the lens) into the immersion medium, and removing liquid from the lens 816 and the surrounding objective lens end and nearby areas (such as, the bottom of the sample support).

[0115] As in other embodiments, the liquid handling device 808 can include a hollow cone 809 including a tapered portion 892 spaced above the end of the objective lens. The cone 809 also includes an annular rim 825 that is coaxial with the longitudinal axis (through the center of the lens 816) and surrounds the lens 816. The rim 825 defines a device opening 827 that exposes the lens 816.

[0116] As in other embodiments, the liquid handling device 808 can generally include a delivery inlet 872 and a return outlet 814, which, as described herein, can be implemented as fluid fittings that couple to the internal passages of the liquid handling device 808. In addition, the delivery inlet 872 and the return outlet 814 can be coupled to a fluid system, such as described above and in Figure 2 A liquid immersion microscope objective system 200 is shown in FIG.

[0117] The liquid delivery system includes a delivery inlet 872, a delivery channel 835, and a delivery outlet 880 in open communication with the lens 816. The components of the liquid delivery system cooperatively define a liquid delivery path from the delivery inlet 872, through the delivery channel 835, through the delivery outlet 880, to the lens 816. In this embodiment, and as Figure 8 and Fig. 9 As shown, the delivery channel 835 can be a single conduit (e.g., formed in or extending through the structure of the liquid handling device 808) that interconnects the delivery inlet 872 and the delivery outlet 880. In addition, the delivery outlet 880 can be a single port adjacent to the lens 816. Therefore, the delivery outlet 880 and the recess 829 are located at different angular positions relative to the center of the lens 816. Specifically, the delivery outlet 880 can be located on the side of the lens 816 opposite the recess 829.

[0118] As described in other embodiments, as described above in conjunction with Figure 2 As mentioned above, the liquid path defined by the liquid delivery system may also be used as a liquid suction (removal) path.

[0119] and Figure 6 and Figure 7 Similar to the illustrated embodiment, the rim 825 includes a notch 829, and the liquid handling device 808 also includes a liquid return passage (or overflow) 831 and a basin or bucket 833. In addition, the width of the return passage 831 can increase along the length of the return passage 831 in a direction away from the notch 829, and the size of the basin 833 can be larger than the size of the notch 829, the return passage 831, and the return inlet 806.

[0120] and Figure 6 and Figure 7 Unlike the embodiment shown in , the return passage 831 and the concave basin 833 are internal features of the structure of the liquid handling device 808. For example, in this embodiment, the return passage 831 is positioned or formed on the inner surface of the conical portion 892 (i.e., the surface inside the conical portion 892), rather than the outer surface of the conical portion 892 (i.e., the surface outside the conical portion).

[0121] Thus, the liquid return system includes a notch 829 in open communication with the lens 816 (e.g., the space above the outer surface of the lens 816 surrounded by the rim 825), a return passage 831, a basin 833, a return inlet 806, and a return outlet 814. The components of the liquid return system cooperatively define a liquid return path from the lens 816, along the return passage 831, into the basin 833, into the return inlet 806, and to the return outlet 814.

[0122] In some embodiments, the return inlet 806 and the return outlet 814 can be part of the same fluid component (eg, fitting). A return passage (eg, internal conduit) can be provided to fluidly interconnect the return inlet 806 and the return outlet 814 as desired.

[0123] Objective lens assembly 800 may otherwise be similar to that described above and as described above. Figures 3 to 5 The objective lens assembly 300 shown in FIG. For example, the objective lens assembly 800 may include the leakage sensor described herein. The objective lens assembly 800 may be used with a sensor such as the one described above and Figure 2 The system is used in combination with the liquid immersion microscope objective system 200 shown in FIG.

[0124] Fig.10 is a schematic cross-sectional front view of a liquid immersion microscope objective lens assembly 1000 according to another embodiment of the present disclosure. Fig.11 yes Fig.10 A top view of the objective lens assembly 1000 is shown in FIG.

[0125] As in other embodiments, the objective lens assembly 1000 generally includes an objective lens 1004 and a liquid handling device 1008 disposed on the longitudinal axis of the objective lens 1004. In addition, the objective lens 1004 includes a lens 1016 mounted at or near the housing opening of the objective lens 1004, so that the lens 1016 is exposed through the housing opening. As in other embodiments, the liquid handling device 1008 is mounted or positioned at the upper portion of the objective lens 1004 and coaxially surrounds the upper portion, as disclosed in other parts of the present disclosure. In addition, the liquid handling device 1008 includes a liquid delivery system and a liquid return system as described herein. Therefore, the liquid handling device 1008 is configured to perform various functions described herein, including immersing the lens 1016 (particularly the outer surface of the lens) into the immersion medium, and removing liquid from the lens 1016 and the surrounding objective lens end and nearby areas (such as, the bottom of the sample support).

[0126] As in other embodiments, the liquid handling device 1008 can include a hollow cone 1009 including a tapered portion 1092 spaced above the end of the objective lens. The cone 1009 also includes an annular rim 1025 coaxial with the longitudinal axis (through the center of the lens 1016) and surrounding the lens 1016. The rim 1025 defines a device opening 1027 that exposes the lens 1016.

[0127] As in other embodiments, the liquid handling device 1008 can generally include a delivery inlet 1072 and a return outlet 1014, which, as described herein, can be implemented as fluid fittings that couple to the internal channels of the liquid handling device 1008. In addition, the delivery inlet 1072 and the return outlet 1014 can be coupled to a fluid system, such as described above and in Figure 2 A liquid immersion microscope objective system 200 is shown in FIG.

[0128] The liquid delivery system includes a delivery inlet 1072, a delivery channel 1035, and a delivery outlet 1080 in open communication with the lens 1016. The components of the liquid delivery system cooperatively define a liquid delivery path from the delivery inlet 1072, through the delivery channel 1035, through the delivery outlet 1080, and to the lens 1016. In this embodiment, and as Fig.10 and Fig.11 As shown, the delivery channel 1035 can be a single conduit (eg, formed in or extending through the structure of the liquid handling device 1008 ) interconnecting the delivery inlet 1072 and the delivery outlet 1080 . Additionally, the delivery outlet 1080 can be a single port adjacent to the lens 1016 .

[0129] The liquid return system includes a return inlet 1006, a return channel 1037, and a return outlet 1014, wherein the return inlet 1006 is in open communication with the lens 1016. The components of the liquid return system cooperatively define a liquid return path from the lens 1016, into the return passage 1006, through the return channel 1037, and to the return outlet 1014. In this embodiment, the construction of the liquid return system is the same or similar to that of the liquid delivery system. Fig.10 and Fig.11 As shown, the return channel 1037 can be a single conduit (e.g., formed in or extending through the structure of the liquid handling device 1008) that interconnects the return inlet 1006 and the return outlet 1014. In addition, the return inlet 1006 can be a single port adjacent to the lens 1016. Therefore, the delivery outlet 1080 and the return inlet 1006 are located at different angular positions relative to the center of the lens 1016. Specifically, the delivery outlet 1080 and the return inlet 1006 can be located on opposite sides of the lens 1016.

[0130] exist Fig.11 , arrows depict the general direction of fluid flow into and out of lens 1016 based on the configuration just described. However, in this embodiment, objective lens assembly 1000 (and associated fluid systems, such as those described above and Figure 2 The liquid immersion microscope objective system 200 shown in FIG. 1 can be configured so that the direction of liquid flow is reversible on either side of the lens 1016. Thus, the liquid delivery system described above can selectively operate the liquid return (or removal, or recovery) system (e.g., by applying a vacuum to the liquid delivery side), and the liquid return system described above can selectively operate the liquid delivery system. For example, when changing the direction of movement of the objective lens assembly 1000 (e.g., when moving from one sample container 132 to another sample container, or from one microplate 128 to another microplate, etc.), the ability to reverse the direction of liquid flow (and thus switch the roles of the delivery outlet 1080 and the return inlet 1006) may be useful.

[0131] Objective lens assembly 1000 may be similar in other respects to that described above and as Figures 3 to 5 The objective lens assembly 300 shown in FIG. For example, the objective lens assembly 1000 may include the leakage sensor described herein. The objective lens assembly 1000 may be used with a sensor such as the one described above and Figure 2 The system is used in combination with the liquid immersion microscope objective system 200 shown in FIG.

[0132] Figure 12 to Figure 14 An example of a liquid immersion microscope objective assembly 1200 according to another embodiment of the present disclosure is shown. Fig.12 is a perspective view of objective lens assembly 1200. Fig.13 yes Fig.12 A cross-sectional front view of the objective lens assembly 1200 is shown in FIG. Fig.14 yes Fig.12 A perspective view of the bottom side of the cone 1209 of the objective lens assembly 1200 is shown. The objective lens assembly 1200 and its operation can be similar to one or more other embodiments described herein. Therefore, the objective lens assembly 1200 generally includes an objective lens 1204 and a liquid handling device 1208 arranged on the longitudinal axis L of the objective lens 1204.

[0133] Objective lens 1204 includes an objective lens housing 1212 and a lens 1216. Housing 1212 extends from a first (upper) end (or objective lens end) to a second (lower) end along a longitudinal axis L, with a first (upper) housing opening 1220 located at the first end and a second (lower) housing opening 1224 located at the second end. Lens 1216 is mounted at or near the first housing opening 1220 so that the outer surface of lens 1216 faces away from objective lens housing 1212 (i.e., facing the area outside housing 1212 and above housing 1212). Therefore, lens 1216 is exposed through first housing opening 1220.

[0134] As in other embodiments, the liquid handling device 1208 is mounted or positioned on the upper portion of the objective lens 1204 and coaxially surrounds the upper portion, as disclosed in other parts of this disclosure. In addition, the liquid handling device 1208 includes a liquid delivery system and a liquid return system as described herein. Therefore, the liquid handling device 1208 is configured to perform various functions described herein, including immersing the lens 1216 (particularly the outer surface of the lens) in the immersion medium, and removing liquid from the lens 1216 and the surrounding objective lens end and nearby areas (such as, The bottom of the sample support).

[0135] Similar to Figures 3 to 5 In the illustrated embodiment, the structure of the liquid handling device 1208 may include a hollow cone 1209 and an annular cover 1226, which cooperatively define a liquid delivery system and a liquid return system. The cone 1209 includes a (first) cone portion 1292 spaced above the end of the objective lens, and a flange portion 1213 adjacent to the cone portion 1292. An annular spacing member 1217 may be placed between the cover 1226 and the flange portion 1213. These components may be fixed together in a stacked arrangement by suitable fasteners such as threaded members (such as screws, bolts, etc.). The spacing member 1217 may be composed of an electrically insulating material, for example.

[0136] The cone 1209 also includes an annular rim 1225 that is coaxial with the longitudinal axis L and surrounds the lens 1216. The rim 1225 defines a device opening that exposes the lens 1216. The rim 1225 transitions to the tapered portion 1292. In this embodiment, the rim 1225 has one or more discontinuities in its geometry, such as a liquid delivery notch 1235 and a liquid return notch 1229 as will be further described below.

[0137] As in other embodiments, the liquid handling device 308 may generally include a delivery inlet 1272 ( Fig.13 ) and return outlet 1214, the delivery inlet 1272 and the return outlet 1214 can be implemented as fluid fittings coupled to internal passages in the structure of the liquid handling device 1208. The delivery inlet 1272 and the return outlet 1214 can be coupled to a fluid system, such as described above and in Figure 2 A liquid immersion microscope objective system 200 is shown in FIG.

[0138] The liquid delivery system includes a delivery inlet 1272, a delivery channel, and a delivery outlet 1280 in open communication with the lens 1216. The components of the liquid delivery system cooperatively define a liquid delivery path from the delivery inlet 1272, through the delivery channel, through the delivery outlet 1280, to the lens 1216. The delivery channel can include one or more conduits (e.g., channels, cavities, etc.) as needed to interconnect the delivery inlet 1272 and the delivery outlet 1280. In this embodiment, as shown in FIG. Fig.13 As shown, the delivery channel includes an annular tapered space 1376, which is defined between the outer surface of the (second) tapered portion 1325 of the objective lens housing 1212 and the inner surface of the tapered portion 1292 of the cone 1209. The tapered space 1376 terminates at a delivery outlet 1280. In this embodiment, the delivery outlet 1280 is an annular opening defined between the edge 1225 and the first housing opening 1220, so that the delivery outlet 1280 coaxially surrounds the first housing opening 1220 and the lens 1216.

[0139] Also in this embodiment, the delivery channel includes a delivery recess 1235, which is in communication with the tapered space 1376 and in open communication with the lens 1216. Fig.14As shown, the delivery channel may include a liquid delivery passage 1437 that is connected to the delivery recess 1235. For example, the delivery recess 1235 may correspond to the end of the delivery passage 1437 where the edge 1225 is located. The delivery passage 1437 may be formed in or on the inner surface of the tapered portion 1292 of the cone 1209. With this configuration, at least a portion of the delivery channel between the delivery inlet 1272 and the delivery outlet 1280 (i.e., the tapered portion 1292) is defined with a main flow path 1439 and a secondary flow path 1441, the main flow path 1439 extending through the delivery passage 1437 to reach the delivery recess 1235, and the secondary flow path 1441 passing through the tapered space 1376 to reach the delivery outlet 1280 (but outside the delivery passage 1437 and the delivery recess 1235). As shown in FIG. Fig.14 As shown, the direction of the secondary flow path 1441 has a circumferential component.

[0140] like Fig.13 As shown, the delivery channel may also include an internal conduit 1329 formed in the cone 1209. The internal conduit 1329 extends from another conduit or cavity 1333 (e.g., formed in the cone 1209) that communicates with the delivery inlet 1272 to an opening in the wall 1337 of the cone 309, which in turn communicates with the tapered space 376. Fig.14 As shown, the opening may correspond to the end of the delivery passage 1437 opposite the delivery recess 1235 .

[0141] As above combined Figure 2 As mentioned above, the liquid path defined by the liquid delivery system may also be used as a liquid suction (removal) path.

[0142] The liquid return system includes a return inlet 1206, a return channel, and a return outlet 1214. The return inlet 1206 is connected to the lens 1216 via the tapered portion 1292. Also in this embodiment, the delivery channel includes a return recess 1229, and the return recess 1229 is in open communication with the lens 1216. Fig.12As shown, the liquid return system may also include a liquid return passage 1231 (or overflow channel) extending from the return recess 1229 to the return inlet 1206. For example, the return recess 1229 may correspond to the start point where the edge 1225 of the return passage 1231 is located. The return passage 1231 may be formed in or on the outer surface of the tapered portion 1292 of the cone 1209. Therefore, in this embodiment, the components of the liquid return system cooperatively define a liquid return path from the lens 1216, into and through the return recess 1229, along or through the return passage 1231, through the return inlet 1206, through the return channel, and to the return outlet 1214. The return recess 1229 and the return passage 1231 may be used as and provide a combination as described above. Figure 6 and Figure 7 However, in this embodiment, the size (particularly the width) of the return passage 1231 can be constant along the length. For example, the width of the return passage 1231 can remain the same (or substantially the same) as the width of the return recess 1229 along the length of the return passage 1231.

[0143] like Fig.12 As shown, the liquid return system can also include a concave basin (or bucket) 1233 surrounding the return inlet 1206. In such an embodiment, the tapered portion 1292 defines a primary flow path 1439 extending from the return recess 1229, through the return passage 1231, and into the return inlet 1206. The concave basin 1233 can collect secondary fluid along the tapered portion 1292 outside the return inlet 1206.

[0144] As is apparent from the foregoing description, the present disclosure provides, in one or more embodiments, a microscope objective assembly (and related systems and methods) that achieves liquid immersion of an objective lens in an efficient manner. The objective assembly also efficiently controls the provision of immersion liquid to the objective lens and the surrounding objective lens end, and removes immersion liquid from the objective lens and the surrounding objective lens end. In addition to generating and maintaining liquid clumps under controlled conditions in the gap between the objective lens and the bottom side of a microplate or other sample support, the objective assembly can remove immersion liquid from both the bottom side of a sample support (such as a microplate, etc.) and the objective lens end, and dry both the bottom side of the sample support and the objective lens end, as required and in response to detected conditions. In addition, the objective assembly can prevent immersion liquid from leaking from the objective lens end onto sensitive components of the microscope's objective assembly operation. In addition, the objective assembly is easily suitable for use in an automated microscope. The function and operation of the objective assembly can be automated, and can therefore be easily integrated into the programmed method development of a given automated microscope and the sample analysis performed by such a microscope. For example, the objective assembly is able to support high-speed and high-throughput imaging of multiple samples, including long-term measurements of samples over hours or days without user intervention.

[0145] It should be understood that terms such as "in communication" and "in communication with..." (e.g., a first component is "in communication" or "in communication with" a second component) used herein indicate a structural, functional, mechanical, electrical, signal, optical, magnetic, electromagnetic, ionic, or fluid relationship between two or more components or elements. Therefore, the fact that one component is described as being in communication with a second component does not exclude the possibility that additional components are present between the first and second components and / or are operably associated or engaged with the first and second components.

[0146] It should be understood that changes may be made to various aspects and details of the present invention without departing from the scope of the present invention. Furthermore, the foregoing description is intended for illustrative purposes only and is not intended to be limiting, the present invention being defined by the claims.

Claims

1. An objective lens assembly, comprising: An objective lens, comprising a housing and a lens mounted on the housing, wherein the lens comprises a lens outer surface; as well as A liquid handling device, which is installed at the objective lens and is configured to immerse the outer surface of the lens in liquid, the liquid handling device comprising: a rim surrounding the outer surface of the lens and defining a device opening for exposing the lens, the rim including a notch; a tapered portion including a tapered surface communicating with the recess; a delivery outlet communicating with the device opening; and a return inlet communicating with the conical surface, in: The liquid handling device defines a liquid delivery path from the delivery outlet to the device opening; and the liquid handling device defining a liquid return path from the device opening, through the recess, along the tapered surface, and to the return inlet, The notch is formed on the tapered portion, and The recess is adapted to capture liquid from the lens, draw the liquid into the recess, direct the liquid along the outer surface of the tapered portion on the side of the lens where the recess is located, and allow the liquid to enter the return inlet located on the same side of the lens as the recess.

2. The objective lens assembly according to claim 1, wherein: The delivery outlet surrounds the lens.

3. The objective lens assembly according to claim 2, wherein: The housing includes a housing opening exposing the lens, the device opening surrounds the housing opening, and the delivery outlet is an annular space defined between the housing opening and the lens opening.

4. The objective lens assembly according to claim 1, wherein: The liquid treatment device includes a delivery channel communicating with the delivery outlet.

5. The objective lens assembly according to claim 4, wherein: The delivery channel includes an annular tapered space.

6. The objective lens assembly according to claim 5, wherein: The tapered portion of the liquid handling device is a first tapered portion, the objective lens includes a second tapered portion, and the annular tapered space is defined between the first tapered portion and the second tapered portion.

7. The objective lens assembly according to claim 1, wherein: The liquid handling device includes a return passage extending from the recess toward the return inlet.

8. The objective lens assembly according to claim 7, wherein: The return passage is disposed on the tapered surface.

9. The objective lens assembly according to claim 8, wherein: The tapered surface is an outer surface outside the tapered portion.

10. The objective lens assembly according to claim 8, wherein: The tapered surface is an inner surface inside the tapered portion.

11. The objective lens assembly according to claim 7, wherein: The return passage has a length between the notch and the return inlet, and a width transverse to the length, and the width increases along the return passage in a direction away from the notch.

12. The objective lens assembly according to claim 7, wherein: The liquid handling device includes a basin in communication with the return inlet, and the return passage extends from the recess toward the basin.

13. The objective lens assembly according to claim 12, wherein: The return passage is disposed on the tapered surface, and the basin is positioned adjacent to the tapered surface.

14. The objective lens assembly according to claim 1, wherein: The delivery outlet includes a port, and the port and the recess are located at different angular positions relative to the center of the lens.

15. The objective lens assembly according to claim 1, wherein: The notch of the liquid return path is a return notch, the rim further comprises a delivery notch, and the liquid delivery path comprises a primary flow path through the delivery notch to the device opening and a secondary flow path through the delivery outlet to the device opening.

16. The objective lens assembly according to claim 15, comprising a delivery passage, the delivery passage communicating with the delivery recess, wherein: The main flow path of the liquid delivery path extends from the delivery passage to the delivery recess.

17. The objective lens assembly according to claim 16, wherein: The delivery passage is provided on or in the inner surface of the tapered portion.

18. The objective lens assembly according to claim 16, wherein: The liquid handling device includes an annular tapered space, and the secondary flow path of the liquid delivery path extends to the device opening through the annular tapered space.

19. The objective lens assembly according to claim 1, comprising a liquid leakage sensor, wherein the liquid leakage sensor is arranged on the liquid handling device.

20. The objective lens assembly according to claim 19, wherein: The liquid leakage sensor is disposed at a portion of the liquid handling device that has a larger radial distance from the longitudinal axis than a radial distance from the tapered surface to the longitudinal axis.

21. The objective lens assembly according to claim 19, wherein: The liquid handling device includes an outer side, and the liquid leakage sensor includes an electrode disposed on the outer side.

22. A method of immersing a lens of a microscope objective with a liquid, the method comprising: Positioning the microscope objective below a sample support such that the lens and the sample support are separated by a gap, wherein the lens is surrounded by a rim and a tapered portion, the rim defining an opening exposing the lens, and the rim including a notch, and the tapered portion including a tapered surface communicating with the notch; flowing liquid to the lens to form and maintain a liquid mass in the gap and contact the liquid mass with the lens and the sample support; and while maintaining the liquid mass, removing the liquid by flowing from the lens into the recess and along a conical surface above the microscope objective, The notch is formed on the tapered portion, and The recess is adapted to capture liquid from the lens, draw the liquid into the recess, direct the liquid along the outer surface of the tapered portion on the side of the lens where the recess is located, and allow the liquid to enter a return inlet located on the same side of the lens as the recess.

23. The method according to claim 22, wherein: Removing liquid also includes flowing the liquid from the recess into a return passage positioned on the tapered surface.

24. The method according to claim 23, wherein: Removing liquid further includes flowing the liquid from the return passage into a basin adjacent the tapered surface.

25. The method according to claim 22, wherein: The sample support includes a first sample container and a second sample container, and the liquid bolus is in contact with the first sample container, and the method further includes: moving at least one of the lens and the sample support relative to the other to position the lens below the second sample container; and While moving, continue to flow or maintain the liquid to keep the lens wet.

26. The method according to claim 25, wherein: Flowing liquid to the lens to form and maintain the liquid mass is performed at a first flow rate, and the method further comprises: Prior to or while moving, the flow of the liquid is adjusted to a second flow rate that is less than the first flow rate.

27. The method according to claim 25, wherein: The liquid bolus in contact with the first sample container is a first liquid bolus, and the method further comprises: After the movement, a second liquid mass is formed in contact with the lens and the second sample container.

28. The method of claim 22, comprising: The liquid is caused to flow from the tapered surface to the return inlet, and suction is applied to the return inlet.

29. The method according to claim 28, comprising: The residual liquid is removed from the bottom of the sample support by draining it into the return inlet via the suction.

30. The method of claim 22, wherein: Flowing the liquid to the lens includes flowing the liquid through a delivery outlet surrounding or adjacent to the lens.

31. The method according to claim 30, comprising: After the liquid mass is formed, the flow of liquid to the lens is stopped and suction is applied to the delivery outlet to remove liquid from the lens.

32. The method of claim 30, comprising: After the liquid bolus is formed, the flow of liquid to the lens is stopped, and suction is applied to the delivery outlet to remove residual liquid from the bottom of the sample support.

33. The method of claim 22, wherein: The tapered surface is part of a liquid handling device, and the method further includes operating a liquid leak sensor to determine whether liquid is present on a portion of the liquid handling device that has a greater radial distance to the lens than the tapered surface has to the lens.

34. An objective lens assembly comprising: An objective lens, comprising a housing and a lens mounted on the housing, wherein the lens comprises a lens outer surface; as well as A liquid handling device, which is installed at the objective lens and is configured to immerse the outer surface of the lens in liquid, the liquid handling device comprising: a rim surrounding an outer surface of the lens and defining a device opening through which the lens is exposed; a tapered portion comprising a tapered surface; a delivery outlet communicating with the device opening; a return inlet in communication with the tapered surface; and an annular V-shaped passage communicating with the return inlet, in: The liquid handling device defines a liquid delivery path from the delivery outlet to the device opening; and The liquid handling device defines a liquid return path from the device opening, along the tapered surface, through the annular V-shaped passage, and to the return inlet; A notch is formed on the tapered portion, and The recess is adapted to capture liquid from the lens, draw the liquid into the recess, direct the liquid along the outer surface of the tapered portion on the side of the lens where the recess is located, and allow the liquid to enter the return inlet located on the same side of the lens as the recess.

35. Objective lens assembly according to claim 34, wherein: The delivery outlet surrounds the lens.

36. Objective lens assembly according to claim 35, wherein: The housing includes a housing opening exposing the lens, the device opening surrounds the housing opening, and the delivery outlet is an annular space defined between the housing opening and the lens opening.

37. The objective lens assembly according to claim 34, wherein: The liquid treatment device includes a delivery channel communicating with the delivery outlet.

38. Objective lens assembly according to claim 37, wherein: The delivery channel includes an annular tapered space.

39. Objective lens assembly according to claim 38, wherein: The liquid handling device includes a first tapered portion, the objective lens includes a second tapered portion, and the annular tapered space is defined between the first tapered portion and the second tapered portion.

40. The objective lens assembly according to claim 34, wherein: The annular V-shaped passage includes an annular inner surface and an annular outer surface defining a V-shaped profile, and the annular inner surface and the annular outer surface are spaced apart from each other at an apex of the annular V-shaped passage to define an annular opening corresponding to the return inlet.

41. The objective lens assembly according to claim 34, wherein: The annular V-shaped passage includes an annular inner surface and an annular outer surface defining a V-shaped profile, and an angle between the annular inner surface and the annular outer surface is in a range of 60 degrees to 100 degrees.

42. The objective lens assembly according to claim 34, comprising a liquid leakage sensor, wherein the liquid leakage sensor is arranged on the liquid handling device.

43. An objective lens assembly according to claim 42, wherein: The liquid leakage sensor is disposed at a portion of the liquid handling device that has a larger radial distance from the longitudinal axis than a radial distance from the annular V-shaped passage to the longitudinal axis.

44. The objective lens assembly according to claim 42, wherein: The liquid handling device includes an outer side, and the liquid leakage sensor includes an electrode disposed on the outer side.

45. The objective lens assembly of claim 34, comprising an electrowetting device configured to apply an electrowetting voltage potential to a surface of the liquid handling device to effectively increase the hydrophilicity of the surface.

46. ​​Objective lens assembly according to claim 45, wherein: The annular V-shaped passage includes an annular inner surface and an annular outer surface defining a V-shaped profile, and the electrowetting device includes a voltage source configured to apply a voltage between the annular inner surface and the annular outer surface.

47. A method of immersing a lens of a microscope objective with a liquid, the method comprising: positioning the microscope objective below a sample support such that the lens and the sample support are separated by a gap; flowing liquid to the lens to form and maintain a liquid mass in the gap and contact the liquid mass with the lens and the sample support; as well as while maintaining the liquid mass, removing liquid by flowing from the lens into an annular V-shaped passageway surrounding the lens; A notch is formed on the tapered portion, and The recess is adapted to capture liquid from the lens, draw the liquid into the recess, direct the liquid along the outer surface of the tapered portion on the side of the lens where the recess is located, and allow the liquid to enter a return inlet located on the same side of the lens as the recess.

48. The method of claim 47, wherein: The sample support includes a first sample container and a second sample container, and the liquid bolus is in contact with the first sample container, and the method further includes: moving at least one of the lens and the sample support relative to the other to position the lens below the second sample container; and While moving, liquid continues to flow to and from the lens to maintain the lens wet.

49. The method of claim 48, wherein: Flowing liquid to the lens to form and maintain the liquid mass is performed at a first flow rate, and the method further comprises: Prior to or while moving, the flow of the liquid is adjusted to a second flow rate that is less than the first flow rate.

50. The method of claim 48, wherein: The liquid bolus in contact with the first sample container is a first liquid bolus, and the method further comprises: After the movement, a second liquid mass is formed in contact with the lens and the second sample container.

51. The method of claim 47, comprising: Suction is applied to the annular V-shaped passage.

52. The method of claim 51, comprising: The residual liquid is removed from the bottom of the sample support by draining it into the annular V-shaped passage via the suction.

53. The method of claim 47, wherein: Flowing the liquid to the lens includes flowing the liquid through an annular delivery outlet surrounding the lens.

54. The method of claim 53, comprising: After the liquid bolus is formed, the flow of liquid to the lens is stopped and suction is applied to the annular delivery outlet to remove liquid from the lens.

55. The method of claim 53, comprising: After the liquid bolus is formed, the flow of liquid to the lens is stopped and suction is applied to the annular delivery outlet to remove residual liquid from the bottom of the sample support.

56. The method of claim 47, wherein: The annular V-shaped channel is part of a liquid handling device, and the method further includes operating a liquid leak sensor to determine whether liquid is present on a surface of the liquid handling device that has a greater radial distance to the lens than the radial distance from the annular V-shaped channel to the lens.

57. The method of claim 47, wherein: The annular V-shaped passage is part of a liquid handling device, and the method further comprises applying an electrowetting voltage potential to a surface of the liquid handling device to effectively render the surface hydrophilic by electrowetting.

58. The method of claim 57, wherein: The electrowetting voltage potential is applied to the annular V-shaped passage.

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