Condensation-reduced microscopy system and method

Abstract

A system for sample imaging includes a control unit for delivering conditioned air and a sample chamber that receives the conditioned air from the control unit. The sample chamber includes a chamber housing having an upper surface, a lower surface opposite the upper surface and configured to face an imaging lens, and a wall extending vertically between the upper and lower surfaces. The wall defines an interior volume of the sample chamber. The sample chamber includes a pneumatic actuator unit configured to direct the conditioned air to a target location along the lower surface to prevent or at least reduce condensation buildup on the imaging lens.

Description

[Technical Field]

[0001] FIELD OF THE DISCLOSURE The present disclosure relates to the field of microscopy, and more particularly to the field of components for providing local environmental conditions. [Background technology]

[0002] To date, those skilled in the art have encountered difficulties in designing on-stage incubation chambers that are compatible with a variety of sample containers (e.g., vessel plates) while also creating an airtight seal with various sample containers. This tends to be problematic because the gap between the bottom edge of the vessel plate and the incubation chamber can allow humid gases to escape. The microscope objective lens is often located directly below the portion of the stage on which the incubation chamber sits and is typically at a much lower temperature than the escaping humid gases. In such situations, humid air that escapes from the incubation chamber (e.g., through the gap between the vessel plate and the chamber) and comes into contact with the objective lens tends to condense on the lens, which can cause imaging problems.

[0003] One option for combating condensation is to use a heating jacket to warm the objective lens, raising its dew point above the condensation point. However, such an approach is primarily used with water-immersion optics to prevent the objective lens from cooling the sample; therefore, this solution is not useful for other types of microscopy. Heating jackets also require physical access to the microscope's objective lens area and do not work with microscopes with rotating objective lens turrets. Other options for dealing with fogging include stopping the experiment to dry the objective lens and / or using anti-fogging agents, but the effectiveness of these methods is limited. Therefore, there is a long-felt need in the art for an improved system for incubation chambers that can be used with minimal or no fogging of the objective lens used to observe samples in the incubation chamber. Summary of the Invention

[0004] In fulfilling the stated needs, the present disclosure provides a system for sample imaging, comprising: a sample chamber configured to contain a local environment therein; and a pneumatic actuator, the sample chamber configured such that air urged by the actuator is directed between the sample chamber and an objective lens, the air being directed to reduce or eliminate condensation on the objective lens.

[0005] In a specific aspect, a sample chamber for use with a sample imaging device includes a chamber housing having a first surface and a second surface facing each other along a first direction, the first surface configured to face an imaging lens, the second surface configured to receive a lid having a window, and the first surface spaced apart from the second surface in a direction facing the lens along the first direction. The chamber housing includes a first end wall and a second end wall facing each other along a second direction substantially perpendicular to the first direction. The chamber housing also includes a first side wall and a second side wall facing each other along a third direction substantially perpendicular to the first and second directions, such that the first and second end walls and the first and second side walls substantially enclose an internal volume in the first and second directions. The internal volume is configured to contain a local environment therein. The chamber housing includes an air actuator unit configured to direct conditioned air to a target location along the first surface and spaced from the first surface in a direction facing the lens, the conditioned air being configured to prevent or at least reduce condensation buildup on the imaging lens.

[0006] In certain aspects, a system for sample imaging includes a control unit for delivering conditioned air and a sample chamber that receives the conditioned air from the control unit. The sample chamber includes a chamber housing having an upper surface, a lower surface opposite the upper surface and configured to face the imaging lens, and a wall extending vertically between the upper and lower surfaces. The wall defines an interior volume of the sample chamber. The sample chamber includes a pneumatic actuator unit configured to direct the conditioned air to a target location along the lower surface to prevent or at least reduce condensation buildup on the imaging lens.

[0007] In certain aspects, the sample chamber comprises features for supporting a sample vessel disposed within the sample chamber. In some embodiments, the sample chamber comprises a lower region for engaging the sample vessel. In some embodiments, the lower region comprises a first heating element. In some embodiments, the first heating element is configured to heat the sample chamber to a temperature in the range of about 30°C to about 40°C. In some embodiments, the heating element further heats air urged by the pneumatic actuator. In some embodiments, the sample chamber comprises a lid. In some embodiments, the lid comprises a second heating element. In some embodiments, the second heating element is configured to heat the sample chamber to a temperature in the range of about 30°C to about 40°C.

[0008] In certain aspects, the chamber comprises a manifold and an outlet, the manifold configured to direct air urged by the actuator to the outlet. In some embodiments, the outlet is aligned with and proximate to the objective lens such that air exiting the outlet impinges on the objective lens. In some embodiments, the outlet is movable. In some embodiments, the outlet is slidable, rotatable, or both. In some embodiments, the manifold further comprises one or more sensors. In some embodiments, the one or more sensors include a temperature sensor.

[0009] In certain aspects, the system further comprises a control unit that delivers conditioned air to the sample chamber. In some embodiments, the control unit comprises a gas mixing manifold. In some embodiments, the gas mixture in the gas mixing manifold comprises one or more gases selected from oxygen, carbon dioxide, nitrogen, and standard air, where the gases are balanced to a mixture of selected concentrations. In some embodiments, the control unit comprises a pump configured to urge the conditioned air to the fluid inlet of the sample chamber. In some embodiments, the pump is located internal to the control unit. In some embodiments, the conditioned air comprises humidified air or heated humidified air. In some embodiments, the conditioned air comprises dry air or heated dry air. In some embodiments, the humidified air comprises a humidity of about 50% to about 90% humidity. In some embodiments, the mixture of selected concentrations comprises about 5% to about 12% carbon dioxide. In some embodiments, the mixture of selected concentrations comprises up to about 21% oxygen. In some embodiments, the mixture of selected concentrations comprises about 67% to about 95% nitrogen.

[0010] In certain aspects, the systems of the present disclosure as described above are configured to operate such that the lowest temperature among all positions of the sample containers located within the sample chamber is within about 15% of the highest temperature among all positions of the sample containers.

[0011] Also provided is a method that includes operating a system for improved imaging as disclosed herein to (1) reduce or eliminate condensation on the objective lens, (2) maintain a first environment within the sample chamber that differs from an ambient environment outside the sample chamber in one or more of temperature, humidity, and gas mixture composition, or do both (1) and (2).

[0012] In some embodiments, the method further includes operating a system as disclosed herein to maintain a second environment within the sample chamber that differs from an ambient environment outside the sample chamber in one or more of temperature, humidity, and gas mixture composition, the second environment being different from the first environment. In some embodiments, the operating is performed such that during operation, the minimum temperature of all locations of the sample container located within the sample chamber is within about 15% of the maximum temperature of all locations of the sample container.

[0013] In the drawings, which are not necessarily drawn to scale, the same numerals may represent similar components in different views. Similar numerals with different letter suffixes may represent different instances of similar components. The drawings generally illustrate, by way of example, but not by way of limitation, various aspects discussed in this document. For example, the accompanying drawings illustrate certain features of the invention, but these drawings should not be considered limiting or exhaustive because the disclosed technology can vary from what is contained in the drawings. [Brief explanation of the drawings]

[0014] [Figure 1] 1 depicts an exemplary incubator system including a control unit and an incubated sample chamber, according to one embodiment of the present disclosure. [Figure 2] 2 depicts an exploded perspective view of the exemplary control unit shown in FIG. 1, according to one embodiment of the present disclosure. [Figure 3] 2 depicts a rear view of the exemplary control unit shown in FIG. 1, showing the interface panel containing the gas inlet, filter for the internal air pump, and connections for the electronics. [Figure 4A] 2 depicts a partially exploded perspective view of the exemplary sample chamber shown in FIG. 1 showing the chamber housing and the removable lid having an insulating film, according to an embodiment of the present disclosure. [Figure 4B]4B depicts an exploded perspective view of selected components of the exemplary chamber housing shown in FIG. 4A, including a pneumatic actuation unit, according to one embodiment of the present disclosure. [Figure 5A] 4B depicts an enlarged perspective view of a portion of the exemplary sample chamber shown in FIG. 4A, with a portion of the chamber housing removed for illustrative purposes to show a portion of the exemplary pneumatic actuation unit. [Figure 5B] 5B depicts a perspective view of the exemplary air actuation unit shown in FIG. 5A. [Figure 5C] 4B depicts an enlarged perspective view of a portion of the exemplary sample chamber shown in FIG. 4A, showing an air filter unit of the exemplary air actuation unit. [Figure 5D] 4B depicts a perspective view of the exemplary air actuation unit shown in FIG. 4A. [Figure 5E] 5D depicts a perspective cross-sectional view of the air actuation unit shown in FIG. 5D. [Figure 5F] 4B depicts a cross-sectional end view of the air actuation unit disposed relative to adjacent components of the chamber housing shown in FIG. 4A. [Figure 5G] FIG. 5E is a top view of the air actuation unit shown in FIG. 5D. [Figure 6A] 5D depicts a perspective view of a fan for use with the air actuation unit shown in FIG. 5D. [Figure 6B] 5D depicts a perspective view of a fan for use with the air actuation unit shown in FIG. 5D. [Figure 7] 4B depicts a bottom perspective view of the exemplary sample chamber shown in FIG. 4A. [Figure 8A] Photographs showing exemplary demonstrations of problematic condensation buildup on objective lenses in microscopes, which is resolved by embodiments of the systems disclosed herein. [Figure 8B] Photographs showing exemplary demonstrations of problematic condensation buildup on objective lenses in microscopes, which is resolved by embodiments of the systems disclosed herein. [Figure 9] 1 illustrates an exemplary method contemplated by the present invention. [Figure 10] FIG. 1 is a schematic diagram illustrating heated air being directed across an objective lens to remove and / or prevent condensation on the lens, according to one embodiment of the present disclosure. DETAILED DESCRIPTION OF THE INVENTION

[0015] The present disclosure may be understood more readily by reference to the following detailed description of the preferred embodiments and examples included therein.

[0016] Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art. In case of conflict, the present document, including definitions, shall control. Preferred methods and materials are described below; however, methods and materials similar or equivalent to those described herein can be used in practice or testing. The materials, methods, and examples disclosed herein are illustrative only and are not intended to be limiting.

[0017] The singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0018] As used in the specification and claims, the term "comprising" can include the embodiments "consisting of" and "consisting essentially of." As used herein, the terms "comprise(s)," "include(s)," "having," "has," "can," "contain(s)," and variations thereof, are intended to be open-ended, transitional phrases, terms, or words that require the presence of the specified components / steps and permit the presence of others. However, such descriptions should be construed as describing compositions or processes as "consisting of" and "consisting essentially of" the listed components / steps, which permit the presence of only the specified components / steps, along with any possible resulting contaminants, and which exclude other components / steps.

[0019] As used herein, the terms "about" and "approximately" mean that the quantity or value in question may be approximately or nearly the same as another value designated. As used herein, it is generally understood to be a nominal value, indicating a variation of ±10%, unless otherwise indicated or inferred. This term is intended to convey that similar values ​​promote the same results or effects as those described in the claims. That is, it is understood that amounts, sizes, formulations, parameters, and other quantities and characteristics need not be exact, but can be approximated and / or larger or smaller as necessary, reflecting tolerances, conversion factors, rounding, measurement errors, etc., and other factors known to those skilled in the art. In general, a quantity, size, formulation, parameter, or other quantity or characteristic is "about" or "approximate," regardless of whether it is explicitly stated as such. When "about" is used before a quantitative value, it is understood that the parameter also includes the specific quantitative value itself, unless otherwise specified.

[0020] Additionally, unless indicated to the contrary, numerical values ​​should be understood to include numerical values ​​that are the same when reduced to the same number of significant digits, and numerical values ​​that differ from the stated value by no more than the experimental error of conventional measuring techniques of the type described in this application to determine the value.

[0021] All ranges disclosed herein are inclusive of and independent of the recited endpoints. The endpoints of ranges and any values ​​disclosed herein are not limited to the exact ranges or values, but are sufficiently imprecise to include values ​​that approximate those ranges and / or values.

[0022] As used herein, approximation terms may be applied to modify any quantitative expression that can be varied without resulting in a change in the underlying function involved. Thus, values ​​modified by terms such as "about" and "substantially" may not be limited to the precise value specified. In at least some instances, approximation terms may correspond to the precision of an instrument for measuring a value. The modifier "about" should also be considered to disclose a range defined by the absolute values ​​of the two endpoints. For example, the phrase "about 2 to about 4" also discloses the range "2 to 4." The term "about" can refer to plus or minus 10% of the indicated number. For example, "about 10%" can indicate a range of 9% to 11%, and "about 1" can mean 0.9 to 1.1. Other meanings of "about" may be apparent from the context, such as rounding; for example, "about 1" can also mean 0.5 to 1.4. Furthermore, while the term "comprising" should be understood to have the open-ended meaning of "including," this term also includes the closed meaning of the term "consisting of." For example, a composition comprising components A and B can be a composition comprising A, B, and other components, but can also be a composition consisting only of A and B. All documents cited herein are incorporated by reference in their entirety for all purposes.

[0023] 1, an incubator system 100 according to the present disclosure includes a control unit 102 and a sample incubation chamber 106 (also referred to herein as a "sample chamber" 106) connected to the control unit 102. The incubator system 100 may also be referred to herein as an "incubator" 100. The control unit 102 is configured to control and regulate a local environment contained within the sample chamber 106. As shown, the control unit 102 may be connected to at least one inlet 107 of the sample chamber 106 via at least one connector 104.

[0024] Referring now to FIG. 2, the control unit 102 can include a gas mixing manifold 110 that can enable mixing of gases received from one or more sources. In some embodiments, the one or more sources can include one or more external sources, i.e., external to the manifold 110, including, for example, one or more gas tanks, gas generators, or other external gas supplies. In some embodiments, the one or more sources can include one or more sources internal to the manifold 110. The manifold 110 can include multiple gas regulation valves and can be connected to external gas supplies, allowing external gases to be mixed within the manifold to the required experimental conditions. The gas supplies can include, but are not limited to, nitrogen (N), carbon dioxide (CO), oxygen (O), air, etc., including one or more combinations thereof. Valves connected to the external gas supplies can be actuated to allow or stop the passage of external gases to the manifold 110. The manifold 110 can include one or more sensors configured to detect one or more gas levels. In this manner, a user can connect the desired gases to manifold 110 and then, by actuation of valves, blend the gases to achieve a mixture within manifold 110 that results in the required experimental conditions. For example, a user can set the O2 level to a value within the range of about 0% to about 21%, including all intermediate values. Similarly, a user can set the CO2 level to a value within the range of about 0% to about 20%, including all intermediate values.

[0025] The composition of the gas mixture in the manifold 110 can be varied over time, i.e., from a first blend at a first time to a second blend at a second time, which can further allow a user to expose a sample in fluid communication with the gas manifold 110 to different gas conditions at different times. A pump 108, such as an air pump 108, can be used to draw air from outside the control unit and / or gas from the gas mixing manifold 110 of the control unit. The control unit 102 can include a water reservoir 114, which can be located within the control unit. In some cases, the lower portion of the reservoir can be aluminum or another thermally conductive material, and the upper portion can be polycarbonate or other plastic. As a non-limiting example, the lower portion of the reservoir can be a material that efficiently conducts heat, and the upper portion of the reservoir can be transparent to allow a user to view the water level therein. A heater, including, for example, a heater plate 116, can be used to heat the water reservoir 114, which can further produce humidified air. The heater plate 116 may be, by way of non-limiting example, a 60-watt (W) heater and may include a digital temperature sensor. Conditioned air, which may include humidified air obtained from heating at least a portion of the contents of the gas mixing manifold 110 and / or the water reservoir 114, can be delivered to the sample chamber 106 from the control unit 102 via the connector 104 (see FIG. 1 ). For example, humidity within the sample chamber 106 is controlled by one or more valves on the gas mixing manifold 110. A valve is actuated to allow the gas mixture to flow over the water reservoir 114, thereby humidifying the gas mixture. When less humidity is preferred, a valve is at least partially closed and a second valve opens, allowing a certain amount of the gas mixture to bypass the heated water reservoir 114. In some embodiments, the connector 104 may also include a heating element that heats the air and allows a user to deliver air to the sample chamber 106 at a desired temperature.A pump 108 can be located within the control unit 102 and can deliver conditioned air (or unconditioned air) from the control unit 102 to the sample chamber 106 via the connector 104. In some embodiments, the humidified air provides an environment in which the sample medium does not evaporate, or evaporates at a reduced rate, allowing users to perform longer experiments while maintaining sample viability and more accurate sample medium volume levels without evaporation.

[0026] As shown in FIG. 2 , the control unit 102 can include one or more of a variety of sensors, such as, by way of non-limiting example, an oxygen sensor 118 and a carbon dioxide (CO2) sensor 112. The control unit 102 can also include one or more gas filter / regulators 120. The gas filter / regulator 120 can be used, for example, to regulate the delivery of gas from an external gas source to the gas mixing manifold 110 and to remove particulates from the circulating air via filtration. In some embodiments, the filter / regulator removes particulates larger than about 5 μm from the circulating air. The filter / regulator operates to control the pressure of the gas to the manifold 110. In some embodiments, the circulating gas is regulated to a pressure of about 35 psi. The control unit 102 can include one or more sensors that report one or more of the temperature, humidity, and gas content of the conditioned air delivered from the control unit 102 via the connector 104. The sensors can be mounted on, adjacent to, or in fluid communication with the manifold 110.

[0027] FIG. 3 provides an external view illustrating a user interface panel 126 that may be located on the rear of the control unit 102. As shown, the user interface panel 126 of the control unit 102 may include an air filter 126a that filters air drawn in by the air pump 108 shown in FIG. 2. The control unit 102 may include one or more gas inlets 126b, which may be connected to an external gas supply, e.g., a supply of O, N, CO gas, and / or air. The control unit 102 may also include a power supply connection 126c, a data and / or power connection 126d to the sample chamber 106 (see FIG. 1), and a data port 126e, e.g., a USB port, that allows connection between the control unit 102 and an external computer or other control device.

[0028] Referring now to FIG. 4A, a partially exploded top perspective view of the sample chamber 106 is shown in accordance with an exemplary embodiment. As shown, the sample chamber 106 may include a chamber housing 150 defining an interior volume 135 for holding one or more sample vessels having sample media therein. The interior volume 135 may also be referred to herein as the "interior" 135 of the sample chamber 106. The sample chamber 106 also preferably includes a cover or "lid" 122 that couples with the chamber housing 150 and encloses a portion of the interior volume 135. The lid 122 preferably includes a window 124 for viewing the sample media located within the interior volume 135 of the sample chamber 106. The lid 122 is described in more detail below. The chamber housing 150 includes at least one inlet 107, which can receive conditioned air from the control unit 102 via at least one connector 104 (see FIG. 1) for delivery to the interior volume 135.

[0029] The chamber housing 150 has a first surface 152 and a second surface 154 opposite the first surface 152 along the first direction Z. The first surface 152 is configured to face an imaging lens, such as the objective lens 142, along the first direction Z. The second surface 154 is configured to be attached to a lid 122. The lid 122 can be fixably attached to the second surface 154. Alternatively, the lid 122 can be a lift-off lid that is not otherwise fixed to the chamber housing 150.

[0030] The first surface 152 is spaced apart from a second surface 154 of the chamber housing 150 in a direction Z1 facing the lens along a first direction Z, and the second surface 154 is spaced apart from the first surface 152 in a direction Z2 facing away from the lens, opposite the direction Z1 facing the lens. It should be understood that the direction Z1 facing the lens and the direction Z2 facing away from the lens are each unidirectional components of the first direction Z, which is bidirectional. The chamber housing 150 also includes a first end wall 156 and a second end wall 158 that face each other along a second direction X that is substantially perpendicular to the first direction Z. The chamber housing 150 further includes a first side wall 160 and a second side wall 162 that face each other along a third direction Y that is substantially perpendicular to the first and second directions. First and second end walls 156, 158 and first and second side walls 160, 162 substantially enclose an internal volume 135 in a second X-direction and a third Y-direction. The internal volume 135 is configured to contain a local environment therein, preferably an incubated local environment for receiving one or more sample vessels. The sample vessels 106 are configured such that a user can remove the lid 122 from the chamber housing 150, place one or more sample vessels within the internal volume 135, and reposition the lid 122 on the second surface 154, thereby enclosing the internal volume 135 in a direction Z2 away from the lens.

[0031] In the illustrated embodiment, the sample chamber 106 is configured to be positioned on a stage (e.g., a movable x, y stage) of a microscope imaging system. Thus, during use, when the sample chamber 106 is positioned in such an orientation, the first direction Z is a vertical direction, and the second and third directions X, Y are each horizontal directions. In such an embodiment, the direction Z1 facing the lens can be characterized as the “downward” direction Z1, and the direction Z2 facing away from the lens can be characterized as the “upward” direction Z2. When used herein with reference to the illustrated embodiment (e.g., when referring to spatial relationships between various features), it should be understood that directional terms can be used to indicate spatial relationships between various features of the sample chamber 106. For example, the terms “downward,” “below,” “bottom,” “directly below,” and derivatives thereof refer to the downward direction Z1, and the terms “upward,” “top,” “above,” “top,” and “top,” and derivatives thereof refer to the upward direction Z2. As some specific, non-limiting examples, when referring to the illustrated embodiments herein, the first surface 152 of the housing body 150 may also be referred to as the “lower” surface 152, and the second surface 154 of the housing body 150 may also be referred to as the “upper” surface 154. Similar such directional terminology is also used herein to describe other features of the illustrated embodiments. However, in other embodiments, the sample chamber 106 may be adapted such that the first direction Z is offset from vertical during use (and thus one or both of the second and third directions may be offset from horizontal). Unless otherwise stated herein, it should be understood that the foregoing spatial relationships of the various described features also refer to the spatial relationships between the various features in embodiments in which the first direction Z is offset from vertical. As two such examples, the sample chamber 106 may be adapted for use with a microscope imaging system in which the imaging lens faces downward rather than upward, or alternatively faces horizontally rather than vertically.In such alternative configurations, the reader will understand that it is the "bottom" 152 of the chamber housing 150 that faces the imaging lens, even if the lower surface 152 is positioned above the upper surface 154 (in the case of a downward-facing lens), or if the lower surface 152 and upper surface 154 are spaced apart horizontally rather than vertically (in the case of a horizontally facing lens). In other words, the directional terms used herein indicate spatial relationships between various features, and unless otherwise stated herein, those spatial relationships apply regardless of the particular orientation in which the sample chamber 106 is oriented in three-dimensional space.

[0032] 4B, the chamber housing 150 can include a first or "lower" housing body 150a and a second or "upper" housing body 150b that are connectable to one another. In the illustrated embodiment, the lower housing body 150a defines a lower surface 152, and the upper housing body 150b defines an upper surface 154, with the lower and upper housing bodies 150a,b being connectable to one another in a sandwich-like configuration. As shown, the lower and upper housing bodies 150a,b can each include respective first and second body end walls 156a,b, 158a,b and first and second body side walls 160a,b, 162a,b, which combine to form portions of the first and second end walls 156, 158 and first and second side walls 160, 160 of the chamber housing 150 when the lower and upper housing bodies 150a,b are coupled together. The lower and upper housing bodies 150a,b also preferably define central apertures 155a,b, respectively, extending therethrough along the first direction Z. The central aperture 155a of the lower housing body 150a provides an open, unobstructed space between the sample chamber 106 and the imaging lens for obtaining a clear image of the sample medium disposed within the sample chamber 106. The central aperture 155b of the upper housing body 150b provides an opening through which a user can place one or more sample containers into the interior volume 135 of the sample chamber 106 while the lid 122 is removed.

[0033] The lower housing portion 150a also preferably has an interior support surface 153, which may extend rim-like around the inner peripheries of the end walls 156a, 158a and side walls 160a, 162a. The interior support surface 157 may be configured to support various features of the sample chamber 106, as described in more detail below. The lower housing portion 150a also preferably has a platform surface 157, which may be located on the side of the first end wall 156a opposite the central aperture 155a along the second direction X. The platform surface 157 may be configured to support various structural features of the chamber housing 150, such as circuitry (e.g., one or more printed circuit boards (PCBs)) and air delivery components, as described in more detail below. The upper housing portion 150b preferably has a canopy portion 159, which may overlap at least a portion of the platform surface 157 and may be configured to cover some or all of the various structural features supported by the platform surface 157.

[0034] The chamber housing 150 may also include a seat member 150c disposed between the lower and upper housing bodies 150a,b. The seat member 150c has an upper end 164 and a lower end 166 spaced apart from one another along the first direction Z. The seat member 150c also includes first and second member end walls 156c, 158d and first and second member side walls 160c, 162c, which form portions of the first and second end walls 156c, 158d and first and second side walls 160c, 162c of the chamber housing 150 when the seat member 150c is coupled together with the lower and upper housing bodies 150a,b. Interior surfaces 170 of the member end walls 156c, 158c and member side walls 160c, 162c define respective portions of the interior volume 135. The first member end wall 156c can define an aperture 172 for passage of one or more vent holes 128 into the interior volume 135, as described in more detail below. The seat member 150c also includes a sample support surface 168 positioned vertically between the upper end 164 and the lower end 166 and facing upward (i.e., along the upward direction Z2). The sample support surface 168 is configured to hold one or more sample containers 175 disposed within the interior volume 135 of the sample chamber 106 (see FIG. 5F). As shown, the sample support surface 168 can extend around the entire inner circumference of the seat member 150c in a rim-like manner and is spaced inward from the member end walls 156c, 158c and the member side walls 160c, 162c. The seat member 150c defines a central aperture 155c extending through the seat member along the first direction Z. Similar to lower housing body 150a, central aperture 155c of seat member 150c provides an open, unobstructed space between sample chamber 106 and imaging lens 168. Additionally, similar to upper housing body 150b, central aperture 155c of seat member 150c also provides an upper opening through which a user places one or more sample containers within interior volume 135 and onto sample support surface 168. Seat member 150c can be made of a material that provides favorable thermal conductivity and insulation for incubating interior volume 135, including, but not limited to, aluminum, steel, and titanium.In such embodiments, seat member 150c may also be referred to as a "heat spreader" 150c. It should be understood that seat member 150c may optionally be made of virtually any machinable, physically stable, and thermally conductive material.

[0035] The sample chamber 106 includes at least one inlet 107, which can communicate with a chamber manifold 130 configured to distribute air delivered to the inlet 107 to one or more vents 128 of the sample chamber 106. In the illustrated embodiment, the chamber manifold 130 defines the inlet 107 at one end thereof and an internal vent 128 at an opposite end thereof. The internal vent 128 extends through, or at least resides within, an aperture 172 in the first end wall 156c of the seat member 150c and is configured to communicate air received from the inlet 107 with the interior volume 135 of the sample chamber 106.

[0036] Continuing with reference to FIG. 4B, the sample chamber 106 includes various thermal regulation features that can be used to heat the environment within the internal volume 135 to a desired temperature, e.g., to incubate the sample medium disposed therein. For example, the thermal regulation features can be configured to heat the environment within the internal volume 135 to a temperature in the range of about 30°C to about 40°C, more specifically in the range of about 35°C to about 40°C, and more preferably about 37°C. The thermal regulation features can also be used to maintain and / or adjust the temperature within the internal volume 135 as needed. Examples of such thermal regulation features are described below. For example, the sample chamber 106 can include a first heater 138 that can extend around the periphery of the internal volume 135. The first heater 138 can have a base heater portion 144 that extends around the periphery of the internal volume 135 and defines a central aperture 155d. The base heater portion 144 can be configured to underlie and heat the lower end 166 of the seat member 150c as needed to maintain the desired temperature within the internal volume 135. The first heater 138 also preferably has one or more wall heater portions 146 extending upwardly along respective side or end walls of the chamber housing 150. In the illustrated embodiment, the first heater 138 includes a pair of wall heater portions 146 extending upwardly from the base portion 138 along first and second side walls 160 a-c, 162 a-c. The pair of wall heater portions 146 may be configured to heat the side walls 160 c, 162 c of the seat member 150 c to further adjust the temperature of the seat member 150 c as needed to maintain a desired temperature within the interior volume 135.

[0037] The sample chamber 106 can also include an inlet heater 148 for heating air delivered through the chamber manifold 130. The inlet heater 148 can include lower and upper panels 174, 176 that contact the upper and lower portions of the chamber manifold 130. In this manner, the inlet heater 148 can operate together with (or provide redundancy for) the first heater 138 as needed to maintain a desired temperature within the interior volume 135. The first heater 138 and the inlet heater 148 also include circuit features such as flex circuits, tracers, connection pins, headers, etc. for electronically connecting the heaters 138, 148 to the control unit 102, which can include, among other things, a processor that executes computer-readable instructions stored in computer memory to control the operation of the heaters 138, 148. Alternatively, the heaters 138, 148 can be electronically connected to a separate electronic control unit that can be located on-board the sample chamber 106.

[0038] It should be understood that the lid 122 (see FIG. 4A ) may also include one or more thermal regulator features, such as, by way of non-limiting example, an insulating film and / or a conductive heater. The insulating film and / or conductive heater may overlay some or all of the window 124 of the lid 122. Thus, the window 124 may also be characterized as a heater or an insulator. The conductive heater and / or insulator of the lid 122 may be used alone or in conjunction with the first heater 138 and / or the inlet heater 148 to heat the environment within the sample chamber 106 to a desired temperature. The insulating film of the lid 122 may include, by way of non-limiting example, one or more suitable insulating films, such as an indium tin oxide resistive film. The sample chamber 106 also preferably includes one or more sensors that report one or more of the temperature, humidity, and gas content of the environment within the interior volume 135 to the control unit 102, as described in more detail below.

[0039] 4B , the sample chamber 106 includes an air circulator or “actuator” unit 132 for directing air to a target location between the internal volume 135 of the sample chamber 106 and the imaging lens relative to the first direction Z. Such target location is preferably beside and below the lower surface 152 of the housing body 150, which location may be referred to as “below” the sample chamber 106. Preferably, the air actuator unit 132 is configured to direct conditioned air, i.e., air having a conditioned (e.g., heated) temperature, to the target location between the internal volume 135 and the imaging lens. By heating such air and directing it to the target location, the sample chamber 106 can effectively warm or heat the imaging lens so as to prevent or at least reduce the formation of condensation thereon, thereby improving image quality. Additionally or alternatively, the heated air can be directed toward or adjacent to the imaging lens to dry or remove any existing condensation that may have accumulated on the imaging lens, thereby improving image quality. The pneumatic actuator unit 132 includes a pneumatic actuator 180, which may be a fan 180 or other mechanism that acts to draw in air, such as ambient air adjacent the sample chamber 106, and direct the drawn air through the pneumatic actuator unit 132. The pneumatic actuator unit 132 also preferably includes a duct member 182 that directs the drawn air from the pneumatic actuator 180 along one or more channels 184 to one or more exit vents or ports 140 facing the target location. The pneumatic actuator unit 132 may include a pneumatic actuator filter 134, which acts to filter the air drawn into the duct member 182 by the pneumatic actuator 180. As shown, the pneumatic actuator filter 134 may be carried by a filter support 137, which may be coupleable to and decoupleable from the duct member 182. The air actuation unit 132 can reside within a compartment 188 defined by portions of the lower and upper housing bodies 150a,b, such as along the first side walls 160a,b thereof, as in the illustrated embodiment.It should be understood that the pneumatic actuation unit 132 may be removable and / or replaceable from the sample chamber 106, as described in more detail below.

[0040] 5A, a partial view of the sample chamber 106 is shown with the upper housing body 150b removed for visualization purposes to show the pneumatic actuator unit 132. As shown, the pneumatic actuator 180 can draw or otherwise direct air (e.g., external ambient air, indicated by arrow 145) into the duct member 182, particularly through an opening 181 defined in the outer surface 183 of the duct member 182. The housing body 150 preferably defines a vent port 190 adjacent the pneumatic actuator 180 to allow external air to enter the pneumatic actuator unit 132. As shown in FIG. 5B, once drawn into the duct member 182 via the pneumatic actuator 180, the directed air is then directed, as indicated by the arrows, through the duct member 182 toward one or more outlet ports 140, which are preferably located directly below sample containers present within the interior volume 135 of the sample chamber 106, as described in more detail below. 5C, the air actuator filter 134 and filter support 137 can reside within a filter receptacle 192 defined by the chamber housing 150, for example, by both its lower and upper housing bodies 150a,b. The filter receptacle 192 and filter support 137 can be configured to facilitate filter replacement by pulling the filter support 137 upwardly from the receptacle 192, as shown.

[0041] 5D-5E , an inner surface 185 of the duct member 182 is shown, the inner surface 185 facing toward the interior volume 135 of the sample chamber 106. The duct member 182 defines one or more channels 184 that direct the induced air flow to one or more outlet vents or ports 140. In the illustrated embodiment, the duct member 182 defines a single channel 184 extending therethrough, defining a flow path leading toward the one or more outlet ports 140. As shown, the air actuation unit 132 can include an outlet member 194 connectable adjacent the duct member 182, and in particular adjacent a lower surface 196 of the duct member 182. The channel 184 can be characterized as having multiple portions, such as a first or opening portion 184a that can be aligned with the air actuator 180 along the third direction Y, a second or main portion 184b that can extend along one of the wall heater portions 146 of the first heater 138 along the second direction X (see FIG. 4B ), and a third or redirecting portion 184c that redirects the induced air flow behind and below the lower surface 152 of the housing body 150. For illustrative purposes, several exemplary air flow paths are shown in FIG. 5E . The second main portion 184b of the channel 184 can be defined between an inner surface 198 of the inner wall 200 of the duct member 182 and an associated wall heater portion 146 (see FIG. 4B ). The associated wall heater portion 146 can be positioned adjacent to or in contact with the inner surface 185 of the duct member 182. In this manner, air guided within channel 184 is directed along the associated wall heater portion 146 in the second direction X, thereby heating the guided air to a temperature sufficient to prevent or at least reduce the formation of condensation on the imaging lens. In redirecting portion 184c of channel 184, the guided air is directed generally in a third direction Y around a bend located downstream of end surface 202 of inner wall 200. Within redirecting portion 184c downstream of the bend, the guided air is further redirected downward (in direction Z1) by the inner surface 204 of outer wall 206 of duct member 182.The downwardly redirected air passes through a bottom opening 208 (shown in FIG. 5E) of the duct member 182 and an associated bottom opening 210 (shown in FIG. 5F) of the lower housing body 150a and enters an outlet passage 212 defined at least in part by the outlet member 194.

[0042] 5F , the outlet passage 212 may be partially defined by the outlet base surface 214 of the outlet member 194 and further partially defined by the underside 152 of the lower housing body 150a. In the illustrated embodiment, the outlet passage 212 is defined perpendicularly between the outlet base surface 214 of the outlet member 194 and the underside 152 of the lower housing body 150a. As shown, the outlet base surface 214 has a curved portion that redirects induced air passing through the bottom openings 208, 210 substantially along the third direction Y. The outlet base surface 214 also has a flat portion that directs the induced air in the third direction Y along the outlet passage 212 toward a target location below the underside 152 of the chamber housing 150. As shown, the flat portion of the outlet base surface 214 is preferably substantially parallel to the underside 152 of the chamber housing 150. This causes the induced air to exit the exit member 194 in respective airflow directions substantially parallel to the underside 152. Additionally, the exit member 194 preferably includes guide members, such as baffles or fins 216, that protrude into the exit passage 212 and are configured to diffuse the induced air exiting the exit port 140 along respective directions having a directional component along the second direction X, thereby expanding the target position to extend along a larger portion of the underside 152 of the chamber housing 150. This provides a more uniform region of conditioned (e.g., heated) air between the sample chamber 106 and the imaging lens, thereby beneficially enhancing the prevention (or at least reduction) of condensation formation on the imaging lens. As shown in FIG. 5G, the fins 216 are elongated along respective directions D1-D4 that diverge from each other in the downstream direction of the airflow D0, thereby spreading the induced air outward as it exits the exit port 140. As shown in FIG. 5F, the fins 216 can extend upward to contact or at least be proximate to the underside 152 of the chamber housing 150, thereby providing multiple outlet ports 140 between the fins 216 along the second direction X.

[0043] 6A and 6B, exemplary embodiments of a pneumatic actuator 180 for use with the pneumatic actuation unit 132 are shown. In these embodiments, the pneumatic actuator 180 comprises a fan 180 having a front surface 230 (FIG. 6A) configured to be positioned adjacent to the outer surface 183 of the duct member 182. The fan 180 also has a rear surface 232 (FIG. 6B) configured to be adjacent to the inner surface 198 of the inner wall 200 of the duct member 182. It should be appreciated that the fan 180 offers numerous advantages for use with the pneumatic actuation unit 132. One advantage discovered by the inventors is that the fan 180 is effective in preventing or at least reducing condensation buildup and / or fogging on the objective lens when used with sample containers of various sizes and configurations. Another such advantage is that the fan 180 also helps maintain a uniform temperature for the sample containers (and the sample media therein), thus improving the incubation environment for the sample media within the sample chamber 106.

[0044] Thus, according to the above-described embodiments, air, such as ambient air, can be directed to the air actuator unit 132, heated by the heater 138, and the heated air can then be directed out one or more exit vents or ports 140 toward, along, or near an imaging lens (e.g., objective lens 142) located below the sample chamber 106. This heated air can then prevent or at least reduce the buildup of fogging and / or condensation on the imaging lens. It should be understood that the air actuator unit 132 can be configured to direct the heated air directly toward the imaging lens or to a region between the imaging lens and the sample chamber 106.

[0045] 7 , the sample chamber 106 preferably includes at least one sensor, such as a temperature / humidity sensor 136, that monitors the environment of the target location directly below the sample chamber 106, such as the temperature and / or humidity of the target location. The output of the temperature / humidity sensor 136 can be used to control the activity of the pneumatic actuator unit 132, heaters 138, 148, and / or other features of the sample chamber 106 disclosed herein. In this manner, the disclosed incubator 100 can (1) create a locally regulated environment, including, for example, a specified temperature and humidity, inside the sample chamber 106, while (2) reducing or eliminating fogging of an imaging lens, such as the objective lens 142, that is imaging a sample or samples disposed within the sample chamber 106.

[0046] 8A and 8B, photographs are shown of an objective lens 142 having condensation thereon as a result of the temperature difference between the objective lens 142 and a prior art sample container. As shown in these photographs, the objective lens 142 has accumulated a significant amount of condensation thereon, which can significantly interfere with images that can be acquired by the objective lens 142 and can require time and effort to remove according to prior art techniques.

[0047] Referring now to FIG. 9 , a process flow of an exemplary method according to the present disclosure is shown. As shown, in step 1, a user can set a first set of desired conditions for the interior 135 of the sample chamber 106, e.g., temperature, humidity, and gas levels to which the sample in the sample chamber 106 will be exposed. The user can then, for example, in step 2, operate the gas manifold 110, pump 108, and other features of the disclosed incubator 100 in an automated manner to achieve the set of desired conditions. The user can then, in step 3, image the sample located in the interior 135 of the sample chamber 106 after exposure to the set of conditions for a desired period of time. The user can then, in step 4, set a next set of desired conditions, such as environmental conditions, for the interior of the sample chamber 106. For example, the environmental conditions may include a particular temperature, humidity, and / or gas mixture level or composition to which the sample in the sample chamber 106 will be exposed, and then perform the previously described steps as needed.

[0048] 10 , a schematic diagram is shown depicting an exemplary air flow provided by air actuation unit 132, according to another embodiment of the present disclosure. In particular, in this embodiment, ambient air 99 a is induced into air actuation unit 132, passes through the air actuation unit, and is directed out one or more exit ports 140 as exhaust air 99 b in a direction intersecting a microscope objective lens 142 located below sample chamber 106. As with the above embodiment, the induced ambient air 99 a in this embodiment may be heated by heater 138 (see FIG. 4B ) as the air travels through air actuation unit 132 and then directed toward, across, or adjacent to objective lens 142 as exhaust air 99 b to dry or remove any condensation that may have accumulated and / or to prevent the accumulation of any condensation on objective lens 142, thereby improving image quality.

[0049] Advantages of the disclosed design of sample chamber 106 include improved uniformity of heating across an incubated sample located within the chamber. Table 1 provides test data showing an array of temperature measurements taken at various locations within a 96-well sample plate incubated within sample chamber 106 as described herein.

[0050] [Table 1]

[0051] The test results shown in Table 1 demonstrate that relatively uniform heating can be achieved and maintained within a sample plate incubated within the sample chamber 106 of the present disclosure. The highest temperature measured within the sample plate was 37.1°C and the lowest temperature measured was 35.6°C, a difference of 1.5°C. The greater uniformity of heating achieved within the sample chamber 106 of the present disclosure provides for improved experimental designs and experimental results obtained from samples contained within the sample chamber 106, as disclosed herein.

[0052] It should be understood that the sample chamber 106 may be adapted to receive sample vessels of various sizes and shapes as desired.

[0053] It should also be understood that in additional embodiments, the incubator system 100 can be provided in a kit that includes the sample chamber 106 and multiple sample containers (e.g., vessel holders or plates) of different sizes and shapes that can be interchangeable within the sample chamber 106.

[0054] It should be further appreciated that in additional embodiments, the air actuation unit can use compressed air as an alternative to fan 180 to prevent or at least reduce condensation buildup and / or fogging on the imaging lens. In such embodiments, the air actuation unit can include a reservoir of compressed air and a nozzle through which the compressed air can be emitted and directed toward, across, or adjacent to the imaging lens. In this manner, the compressed air can create a pressure differential within a target location between the sample chamber and the imaging lens. Such a pressure differential can be used to lower the dew point temperature at the target location, thereby preventing or at least reducing condensation buildup on the imaging lens and / or fogging of the imaging lens.

[0055] It should be appreciated that, according to further additional embodiments herein, air actuation unit 132 can be said to include means for adjusting air conditions at a target location across, at, or adjacent to the imaging lens. In such embodiments, the means for adjusting air conditions at the target location can include fan 180, compressed air, or other features and techniques. Air actuation unit 132 according to such embodiments can also include means for directing the conditioned air to the target location, which can include a duct member such as duct member 182 described above, and can also include an outlet member such as outlet member 194 described above. The means for directing the conditioned air to the target location can also include fins or baffles, such as fins 216 described above.

[0056] Furthermore, when a numerical preposition (e.g., "first," "second," "third") is used herein in reference to an element, component, dimension, or feature thereof (e.g., a "first" sensor, a "second" sensor), it should be understood that such numerical preposition is used to distinguish that element, component, dimension, and / or feature from another such element, component, dimension, and / or feature and should not be limited to the particular numerical preposition used in that instance. For example, a "first" sensor could also be referred to as a "second" sensor in a different context without departing from the scope of the present disclosure, so long as that element, component, dimension, and / or feature remains appropriately distinguished in the context in which the numerical preposition is used.

[0057] Although the present disclosure has been described in detail, it should be understood that various changes, substitutions, and alterations can be made therein without departing from the spirit and scope of the present invention, as defined by the appended claims. Moreover, the scope of the present disclosure is not intended to be limited to the particular embodiments described herein. In particular, one or more features from the foregoing embodiments can be used in other embodiments herein. As one skilled in the art will readily appreciate, any now-existing or later-developed process, machine, manufacture, composition of matter, means, method, or step that performs substantially the same function or achieves substantially the same result as the corresponding embodiment described herein can be utilized in accordance with the present disclosure.

[0058] Aspects The following aspects are illustrative only and do not limit the scope of the disclosure or the appended claims. Any part or parts of any one or more aspects may be combined with any part or parts of any one or more other aspects.

[0059] Aspect 1. A system for improved imaging, comprising: a sample chamber configured to contain a local environment therein; a pneumatic actuator; The sample chamber is configured such that air urged by the actuator is directed between the sample chamber and an objective lens configured to image the sample in the sample chamber, the air being directed to reduce or eliminate condensation on the objective lens, the system.

[0060] Embodiment 2. The system of embodiment 1, wherein the sample chamber comprises a feature for supporting a sample vessel disposed within the sample chamber.

[0061] Embodiment 3. The system of embodiment 1, wherein the sample chamber comprises a lower region for engaging a sample container.

[0062] Embodiment 4. The system of embodiment 3, wherein the lower region comprises a heating element.

[0063] Embodiment 5. The system of embodiment 4, wherein the heating element is configured to heat the sample chamber to about 37°C.

[0064] Embodiment 6. The system of embodiment 4, wherein the heating element further heats the air urged by the air actuator.

[0065] Embodiment 7. The system of embodiment 1, wherein the sample chamber comprises a lid.

[0066] Embodiment 8. The system of embodiment 7, wherein the lid comprises a heating element.

[0067] Embodiment 9. The system of embodiment 8, wherein the heating element is configured to heat the sample chamber to 37°C.

[0068] Aspect 10. The system of aspect 1, wherein the chamber comprises a manifold and an outlet, the manifold configured to direct air urged by the actuator to the outlet.

[0069] Embodiment 11. The system of embodiment 10, wherein the outlet is aligned with the objective lens.

[0070] Embodiment 12. The system of embodiment 11, wherein the outlet is movable.

[0071] Embodiment 13. The system of embodiment 12, wherein the outlet is slidable, rotatable, or both.

[0072] Embodiment 14. The system of embodiment 10, wherein the manifold further comprises one or more sensors.

[0073] Embodiment 15. The system of embodiment 14, wherein the one or more sensors include a temperature sensor.

[0074] Embodiment 16. The system of embodiment 1, further comprising a control unit that delivers conditioned air to the sample chamber.

[0075] Embodiment 17. The system of embodiment 16, wherein the control unit comprises a gas mixing manifold.

[0076] Aspect 18. The system of aspect 17, wherein the gas mixture in the gas mixing manifold comprises one or more gases selected from oxygen, carbon dioxide, nitrogen, and standard air, and the gases are balanced to a mixture of selected concentrations.

[0077] Embodiment 19. The system of embodiment 16, wherein the control unit comprises a pump configured to urge conditioned air to the fluid inlet of the sample chamber.

[0078] Aspect 20. The system of aspect 19, wherein the pump is positioned within the control unit.

[0079] Aspect 21. The system of Aspect 16, wherein the conditioned air comprises humidified air or heated humidified air.

[0080] Aspect 22. The system of aspect 16, wherein the conditioned air comprises dry air or heated dry air.

[0081] Embodiment 23. The system of embodiment 21, wherein the humidified air comprises a humidity of about 50% to about 90% humidity.

[0082] Embodiment 24. The system of embodiment 18, wherein the mixture of selected concentration comprises about 5% to about 12% carbon dioxide.

[0083] Embodiment 25. The system of embodiment 18, wherein the mixture of selected concentrations comprises up to about 21% oxygen.

[0084] Embodiment 26. The system of embodiment 18, wherein the mixture of selected concentrations comprises between about 67% and about 95% nitrogen.

[0085] Embodiment 27. The system of any one of embodiments 1 to 26, wherein the system is configured to operate such that the minimum temperature among all positions of the sample container located within the sample chamber is within about 15% of the maximum temperature among all positions of the sample container.

[0086] Aspect 28. A method comprising operating a system described in any one of aspects 1 to 27 to: (1) reduce or eliminate condensation on an objective lens; (2) maintain a first environment within the sample chamber that differs in one or more of temperature, humidity, and gas mixture relative to an ambient environment outside the sample chamber; or do both (1) and (2).

[0087] Aspect 29. The method of aspect 28, further comprising operating the system of any one of aspects 1 to 27 to maintain a second environment within the sample chamber that differs in one or more of temperature, humidity, and gas mixture relative to an ambient environment outside the sample chamber, wherein the second environment is different from the first environment.

[0088] Embodiment 30. A method according to any one of embodiments 28 to 29, wherein the operating is performed such that during operation, the minimum temperature at all positions of the sample container located within the sample chamber is within about 15% of the maximum temperature at all positions of the sample container.

[0089] Aspect 31. An incubator comprising: a sample chamber having an interior and (i) a fluid inlet, (ii) a surface for receiving a sample container, and (iii) one or more heating elements; and a control unit configured to engage the sample chamber, wherein the fluid inlet is configured to receive conditioned air from the control unit, and the sample chamber is configured to deliver the conditioned air to an interior of the sample chamber.

[0090] The sample chamber may be placed on a microscope stage and / or a microscope slide. A gasket may be used to seal the sample chamber against the microscope stage and / or slide.

[0091]

[0033] Aspect 32. The incubator of Aspect 31, wherein the control unit includes a gas mixing chamber, also referred to in some cases as a gas mixing manifold. The gas mixing chamber can include any one or more of valves, sensors, filters, and / or regulators to generate the gas mixture for delivery to the fluid inlet. The one or more sensors can be positioned to monitor any one or more of gas content, temperature, and humidity of the contents of the gas mixing manifold.

[0092] Aspect 33. The incubator of Aspect 32, wherein the gas mixture in the gas mixing chamber comprises one or more gases selected from oxygen, carbon dioxide, nitrogen, and standard air, the gases being balanced to a mixture of selected concentrations. One or more such gases can be received from a gas source external to the gas mixing chamber, e.g., a tank of gas. By adjusting the introduction of different gases into the gas mixing chamber, a user can achieve mixing within the gas mixing chamber according to set specifications for gas levels.

[0093] Aspect 34. The incubator of Aspect 31, wherein the conditioned air comprises humidified air or heated humidified air. A user can, for example, set a desired humidity level, which can be achieved by humidifying the air drawn into the control unit, by mixing humidified air generated within the control unit with the air drawn into the control unit, or both. It should be understood that the contents of the gas mixing manifold can be mixed with humidified air and / or non-humidified air.

[0094] Embodiment 35. The incubator of embodiment 31, wherein the conditioned air comprises dry air or heated dry air. The incubator can include a dehumidifier, e.g., a dehumidifier configured to dehumidify the air delivered to the inlet of the sample chamber via the connector.

[0095] Embodiment 36. The incubator of embodiment 31, further comprising a lid.

[0096] Embodiment 37. The incubator of embodiment 36, wherein the lid comprises (i) a thermal insulating film coating, (ii) a conductive heater, or both. The lid can be connected to a power supply, which further acts to heat the lid. The lid can also comprise a power source, such as a battery, which is used to energize the lid heater.

[0097] Embodiment 38. The incubator of embodiment 31, wherein the control unit comprises a pump configured to urge conditioned air to the fluid inlet.

[0098] Aspect 39. An incubator according to aspect 38, wherein the pump is positioned within the control unit.

[0099] Embodiment 40. The incubator of embodiment 34, wherein the humidified air is humidified to a humidity of about 50% to about 90%, preferably about 80%. Humidity levels of 50% to 90%, 55% to 85%, 60% to 80%, or even 70% are all suitable.

[0100] Embodiment 41. The incubator of embodiment 33, wherein the selected mixture comprises between about 5% and about 12% carbon dioxide, preferably about 5% carbon dioxide. Carbon dioxide levels of between about 5% and about 12%, between about 6% and about 11%, between about 7% and about 10%, or even between about 8% and about 9% are all believed to be suitable.

[0101] Embodiment 42. The incubator of embodiment 33, wherein the selected mixture comprises up to about 21% oxygen. The mixture can have less than 21% oxygen, e.g., between about 0.5% and about 20%, between about 1% and about 19%, between about 2% and about 18%, between about 3% and about 17%, between about 4% and about 16%, between about 5% and about 15%, between about 6% and about 14%, between about 7% and about 13%, between about 8% and about 12%, between about 9% and about 11%, or even about 10%.

[0102] Embodiment 43. The incubator of embodiment 33, wherein the selected mixture comprises about 67% to about 95% nitrogen, preferably about 75% nitrogen. The mixture can have about 67% to about 95% nitrogen, or about 70% to about 90% nitrogen, or about 75% to about 85% nitrogen, or even about 80% nitrogen.

[0103] Embodiment 44. The incubator of embodiment 34 or 35, wherein the conditioned air is heated to about 37° C. However, this is not required, as the conditioned air can be heated to, for example, about 20° C., about 25° C., about 30° C., or even about 35° C.

[0104] Embodiment 35. The incubator of embodiment 31, wherein the one or more heating elements are configured to heat the specimen sample container to approximately 37°C.

[0105] Embodiment 46. The incubator of embodiment 31, further comprising a manifold configured to receive conditioned air.

[0106] Embodiment 47. The incubator of embodiment 36, wherein the manifold is configured to distribute conditioned air to the interior of the sample chamber. The manifold can extend around a portion of, or even the entire perimeter (inside or outside) of, the interior of the sample chamber.

[0107] Embodiment 48. The incubator of any one of embodiments 46-47, further comprising at least one vent in fluid communication with the manifold, the at least one vent configured to direct conditioned air received from the control unit to the sample vessels. The conditioned air directed to the sample vessels can be heated by one or more heating elements in the sample chamber.

[0108] Aspect 49. The incubator of Aspect 31, further comprising an air circulation device embodied within the sample chamber, the air circulation device configured to encourage air below the sample chamber. Such air can be used to reduce or even eliminate fogging of an objective lens positioned below the sample chamber.

[0109] Embodiment 50. The incubator of embodiment 49, wherein the air actuator comprises a fan.

[0110] Embodiment 51. The incubator of any one of embodiments 31 to 50, further comprising at least one sensor configured to detect humidity, gas levels, or both.

[0111] Aspect 52. An incubator described in any one of aspects 31 to 51, further comprising a water reservoir, the water reservoir being in fluid communication with the control unit and / or being embodied within the control unit, the water reservoir being in fluid communication with the fluid inlet.

[0112] Aspect 53. An incubator according to any one of aspects 31 to 53, wherein the incubator is configured to include in the conditioned air: (i) a gas mixture, (ii) humidified air, or both (i) and (ii).

[0113] Aspect 54. An incubator according to any one of aspects 31 to 53, wherein the incubator adjusts the atmosphere inside the sample chamber to at least one specified level of any one or more conditions selected from oxygen level, carbon dioxide level, nitrogen level, humidity percentage, or temperature.

[0114] Embodiment 55. The incubator of embodiment 31, wherein the gas mixing chamber mixes gases received from outside the incubator.

[0115] Embodiment 56. An incubator according to embodiment 31, wherein the incubator is configured to be mounted on a microscope stage.

[0116] Embodiment 57. An incubator according to embodiment 31, wherein the chamber comprises one or more ports configured to introduce one or more materials into the interior of the sample chamber or remove one or more materials from the interior of the sample chamber.

[0117] Embodiment 58. A method for imaging a sample in a controlled environment, the method comprising: positioning the sample in a sample chamber of an incubator according to any one of embodiments 31 to 57; and acquiring one or more images of the sample in the sample chamber. The image acquisition can be performed, for example, via an inverted microscope.

[0118] Embodiment 59. The method of embodiment 58, further comprising exposing the sample positioned within the sample chamber to a first set of environmental conditions in an atmosphere within the sample chamber for a first time interval.

[0119] Aspect 60. The method of aspect 59, wherein the first set of environmental conditions includes a first specified level of one or more of oxygen, carbon dioxide, nitrogen, humidity, temperature, or one or more combinations thereof.

[0120] Embodiment 61. The method of embodiment 59, wherein the first set of environmental conditions includes conditions that are different from corresponding conditions of the atmosphere outside the sample chamber.

[0121] Embodiment 62. The method of any one of Embodiments 58 to 61, wherein the first time interval ranges from about 1 minute to 72 hours or more. For example, intervals of about 1 minute to 72 hours or more, about 1 minute to 48 hours, about 5 minutes to 24 hours, about 10 minutes to 20 hours, about 30 minutes to 10 hours, and 1 hour to 5 hours are all considered suitable.

[0122] Embodiment 63. The method of any one of embodiments 58-62, further comprising exposing the sample to a second set of environmental conditions inside the sample chamber for a second time interval.

[0123] Aspect 64. The method of aspect 63, wherein the second set of environmental conditions includes conditions that are different from corresponding conditions in the first set of environmental conditions, e.g., including temperature, humidity, and / or gas mixture. The second time interval can be different from the first time interval.

[0124] Embodiment 65. The method of any one of embodiments 58 to 64, further comprising introducing one or more reagents into the sample chamber.

[0125] Embodiment 66. The method of any one of embodiments 58 to 65, further comprising extracting one or more samples from the sample chamber.

Claims

1. 1. A sample chamber for use with a sample imaging device, comprising: A chamber housing, a first surface and a second surface facing each other along a first direction, the first surface being configured to face an imaging lens, the second surface being configured to mount a lid having a window, and the first surface being spaced apart from the second surface in a direction facing the lens along the first direction; a first end wall and a second end wall opposed to each other along a second direction substantially perpendicular to the first direction; a first sidewall and a second sidewall facing each other along a third direction substantially perpendicular to the first direction and the second direction; a chamber housing, the first and second end walls and the first and second side walls substantially enclosing an interior volume in the first and second directions, the interior volume configured to contain a local environment therein; an air actuator unit configured to direct conditioned air to a target location along the first surface and spaced from the first surface in a direction facing the lens to prevent or at least reduce condensation buildup on the imaging lens.

2. 2. The sample chamber of claim 1, wherein the pneumatic actuator unit comprises a pneumatic actuator, a duct member, and an outlet port, the pneumatic actuator configured to induce air from outside the sample chamber and direct the induced air into a channel defined by the duct member, the channel configured to direct the induced air to the outlet port such that the induced air exits the outlet port and travels outside of and along the second surface of the chamber housing.

3. The sample chamber of claim 2 , wherein the pneumatic actuator comprises a fan.

4. 4. A sample chamber according to claim 2 or 3, wherein the pneumatic actuator unit comprises an outlet member in communication with the duct member, the outlet member at least partially defining an outlet passage terminating in the outlet port.

5. The sample chamber of claim 4 , wherein the outlet member comprises fins projecting into the outlet passage and configured to direct the flow of air exiting the outlet port.

6. 6. The sample chamber of claim 5, wherein the fins are elongated along respective directions that diverge from one another in a downstream direction, thereby spreading the induced air outward as it exits the outlet port.

7. 6. The sample chamber of claim 4, wherein the outlet member has an outlet base surface that at least partially defines the outlet passage, the fins extending from the outlet base surface along a direction opposite to the direction facing the lens, and the outlet base surface being substantially parallel to the first surface of the chamber housing, thereby causing the induced air to exit the outlet member in a respective air flow direction that is substantially parallel to the first surface of the chamber housing.

8. 8. The sample chamber of claim 7, wherein the outlet base surface faces a portion of the first side of the chamber housing, such that the portion of the first side partially defines the outlet passage.

9. A sample chamber according to any preceding claim, further comprising a heating element configured to heat the internal volume of the sample chamber.

10. The sample chamber of claim 9, wherein the heating element is configured to heat the interior volume of the sample chamber to a temperature in the range of about 30°C to about 40°C.

11. 11. A sample chamber as described in claim 9 or 10, wherein at least a portion of the heating element is adjacent to the channel, whereby the induced air is directed along the at least a portion of the heating element and the induced air is heated by the heating element.

12. A sample chamber according to any one of claims 1 to 11, further comprising at least one temperature sensor configured to measure the temperature at the target location.

13. 13. The sample chamber of claim 1, further comprising a lid configured to be repeatedly attached to and detached from the second surface of the chamber housing, the lid comprising at least one of a heating element or a thermal insulating layer.

14. 14. The sample chamber of claim 13, wherein the lid comprises a lid heating element, the lid heating element configured to heat the sample chamber to a temperature in the range of about 30°C to about 40°C.

15. 1. A system for sample imaging, comprising: a control unit for delivering conditioned air; a sample chamber, the sample chamber comprising: a chamber housing having an upper surface, a lower surface opposite the upper surface, and a wall extending vertically between the upper surface and the lower surface, the walls defining an interior volume of the sample chamber, the lower surface configured to face an imaging lens; an air actuator unit configured to direct conditioned air to a target location along the underside to prevent or at least reduce condensation buildup on the imaging lens.

16. 16. The system of claim 15, wherein the pneumatic actuator unit comprises a pneumatic actuator, a duct member, and an outlet port, the pneumatic actuator configured to induce air from outside the sample chamber and direct the induced air into a channel defined by the duct member, the channel configured to direct the induced air to the outlet port such that the induced air exits the outlet port and travels outside of and along the second surface of the chamber housing.

17. 17. The system of claim 16, wherein the air actuator comprises a fan and the sample chamber comprises at least one heater located along the channel, the at least one heater configured to heat the induced air before it exits the outlet port.

18. 18. The system of claim 16 or 17, wherein the outlet ports include fins that are elongated along respective directions that diverge from one another in the downstream direction, thereby spreading the induced air outward as it exits the outlet ports.

19. 19. The system of any one of claims 15 to 18, further comprising a lid configured to be repeatedly attached to and detached from the upper surface of the chamber housing, the lid comprising at least one of a heating element or an insulating layer.

20. 20. The system of claim 19, wherein the lid comprises a lid heating element, the lid heating element configured to heat the sample chamber to approximately 37°C.

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