System and method for reduced coacervation microscopy

Abstract

A system for sample imaging includes a control unit for delivering conditioned air and a sample chamber receiving the conditioned air from the control unit. The sample chamber includes a chamber housing having an upper face, a lower face opposite the upper face and configured to face an imaging lens, and a wall extending vertically between the upper face and the lower face. The wall defines an interior volume of the sample chamber. The sample chamber includes an air actuator unit configured to direct conditioned air to a target location alongside the lower face to suppress or at least reduce aggregate build-up on the imaging lens.

Description

Technical Field

[0001] The present disclosure relates to the field of microscopy, and in particular to the field of components for influencing local environmental conditions. Background Art

[0002] To date, those skilled in the art have encountered difficulties in designing an incubation chamber on an object stage that is compatible with various sample containers (e.g., a vessel plate) and also forms an airtight seal with various sample containers. This often presents a problem because the gap between the bottom edge of the vessel plate and the incubation chamber may allow humid gases to escape. The microscope objective lens is usually directly below the portion of the object stage where the incubation chamber is located and is usually much cooler in temperature than the escaping humid gases. In such instances, humid air that escapes from the incubation chamber (e.g., through the gap between the vessel plate and the incubation chamber) and contacts the objective lens tends to condense on the lens, which may cause imaging problems.

[0003] One option to prevent condensation is to use a heating jacket that heats the objective lens to raise its dew point above the condensation point. However, this method is mainly used with water immersion optics to prevent the sample from being cooled by the objective lens, and therefore this solution is not suitable for other types of microscopy. The heating jacket may also require physical access to the objective lens area of ​​the microscope and will not work on a microscope with a rotating objective turret. Other options for addressing fogging include stopping the experiment to allow the objective lens to dry and / or using anti-fog agents, but these methods have limited effectiveness. Therefore, there has long been a need in the art for improved systems for incubation chambers that can be used with minimal or even no fogging of the objective lens used to observe samples in the incubation chamber. Summary of the Invention

[0004] To address the described needs, the present disclosure provides a system for imaging a sample, the system comprising: a sample chamber configured to contain a local environment therein; and an air actuator, the sample chamber configured such that air encouraged by the actuator is directed between the sample chamber and an objective lens, the air being directed so as to reduce or eliminate condensation on the objective lens.

[0005] In certain aspects, a sample chamber for use with a sample imager includes a chamber housing having a first face and a second face opposing each other along a first direction, wherein the first face is configured to face an imaging lens, the second face is configured to receive a cover having a window, and the first face and the second face are spaced apart in a lens-facing direction along the first direction. The chamber housing includes a first end wall and a second end wall opposing 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 opposing 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 interior volume relative to the first and second directions. The interior 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 located adjacent to the first face and spaced apart from the first face in the lens-facing direction. The conditioned air is configured to inhibit or at least reduce condensation accumulation on the imaging lens.

[0006] In certain aspects, a system for imaging a sample 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 face, a lower face opposite the upper face and configured to face an imaging lens, and walls extending vertically between the upper face and the lower face. The walls define an interior volume of the sample chamber. The sample chamber includes an air actuator unit configured to direct the conditioned air to a target location located adjacent to the lower face to inhibit or at least reduce condensation accumulation on the imaging lens.

[0007] In some aspects, the sample chamber includes features for supporting a sample container disposed within the sample chamber. In some embodiments, the sample chamber includes a lower region for engaging with a sample container. In some embodiments, the lower region includes a first heating element. In some embodiments, the first heating element is configured to heat the sample chamber to a range of about 30°C to about 40°C. In some embodiments, the heating element further heats the air excited by the air actuator. In some embodiments, the sample chamber includes a lid. In some embodiments, the lid includes a second heating element. In some embodiments, the second heating element is configured to heat the sample chamber to a range of about 30°C to about 40°C.

[0008] In some aspects, the chamber includes a manifold and an outlet, wherein the manifold is configured to direct air excited by the actuator to the outlet. In some embodiments, the outlet is aligned with and proximate to the objective lens so 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 includes 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, and these gases are balanced to a selected concentration mixture. In some embodiments, the control unit comprises a pump that is configured to excite the fluid inlet of the conditioned air to the sample chamber. In some embodiments, the pump is positioned inside 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 has a humidity of about 50% to about 90% humidity. In some embodiments, the selected concentration mixture comprises about 5% to about 12% carbon dioxide. In some embodiments, the selected concentration mixture comprises up to about 21% oxygen. In some embodiments, the selected concentration mixture comprises about 67% to about 95% nitrogen.

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

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

[0012] In some embodiments, the method further comprises operating a system disclosed herein to maintain a second environment within the sample chamber, the second environment being different 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 operation is performed such that during the operation, a minimum temperature of all positions of the sample container within the sample chamber is within about 15% of a maximum temperature of all positions of the sample container. BRIEF DESCRIPTION OF THE DRAWINGS

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

[0014] In the attached figure: Figure 1 An exemplary incubator system including a control unit and an incubation sample chamber according to embodiments of the present disclosure is depicted.

[0015] Figure 2 Depicted is an embodiment of the present disclosure Figure 1 An exploded perspective view of an exemplary control unit is illustrated.

[0016] Figure 3 Depicts Figure 1 A rear view of an exemplary control unit is illustrated showing the interface panel including the gas inlet, filter for the internal air pump, and connections for the electronics.

[0017] Figure 4A Depicted is an embodiment of the present disclosure Figure 1 A partially exploded perspective view of an exemplary sample chamber is illustrated showing the chamber housing and removable cover with thermal insulation membrane.

[0018] Figure 4B Depicted is an embodiment of the present disclosure Figure 4A An exploded perspective view of selected components of an exemplary chamber housing, including an air actuation unit, is illustrated.

[0019] Figure 5A Depicts Figure 4A An enlarged perspective view of a portion of an exemplary sample chamber is illustrated with a portion of the chamber housing removed for illustrative purposes to show a portion of an exemplary air actuation unit.

[0020] Figure 5B Depicts Figure 5A A perspective view of an exemplary air actuation unit is illustrated.

[0021] Figure 5C Depicts Figure 4A An enlarged perspective view of a portion of an exemplary sample chamber is illustrated showing an air filter unit of an exemplary air actuation unit.

[0022] Figure 5D Depicts Figure 4A A perspective view of an exemplary air actuation unit is illustrated.

[0023] Figure 5E Depicts Figure 5D A perspective cutaway view of an illustrated air actuation unit.

[0024] Figure 5F Depicts relative to Figure 4A End cross-sectional view of an air actuation unit shown with adjacent components of a chamber housing.

[0025] Figure 5G yes Figure 5D Top view of an illustrated air actuation unit.

[0026] Figure 6A and Figure 6B Depicts the use of Figure 5D A perspective view of a fan for use with the illustrated air actuation unit.

[0027] Figure 7 Depicts Figure 4A A bottom perspective view of an exemplary sample chamber is illustrated.

[0028] Figure 8A and Figure 8B is a photograph showing an exemplary demonstration of problematic condensate accumulation on an objective lens in a microscope, which is addressed by embodiments of the system disclosed herein.

[0029] Figure 9 Exemplary methods contemplated by the present invention are depicted.

[0030] Figure 10 Depicted is a schematic diagram showing heated air directed through an objective lens to remove and / or prevent condensation on the lens, in accordance with an embodiment of the present disclosure. DETAILED DESCRIPTION

[0031] The present disclosure may be understood more readily by reference to the following detailed description of desired embodiments and the Examples included therein.

[0032] 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 the event of a conflict, the present document (including definitions) will prevail. Although methods and materials similar or equivalent to those described herein can be used in practice or testing, preferred methods and materials are described below. The materials, methods, and examples disclosed herein are illustrative only and not restrictive.

[0033] The singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise.

[0034] As used in the specification and claims, the term "comprising" may include the embodiments of "consisting of" and "consisting essentially of. As used herein, the terms "comprising," "including," "having," "may," "containing," and variations thereof are intended to be open-ended transitional phrases, terms, or words that require the presence of a specified ingredient / step and permit the presence of other ingredients / steps. However, such descriptions should be understood as also describing a composition or method as "consisting of" and "consisting essentially of the enumerated ingredients / steps," permitting the presence of only the recited ingredients / steps, along with any impurities that may result therefrom, and excluding other ingredients / steps.

[0035] As used herein, the terms "about" and "equal to or approximately" mean that the amount or value in question can be a value specified as being approximately or roughly the same as some other value. As used herein, it is generally understood to be a ±10% variation of the specified nominal value, unless otherwise stated or inferred. The terms are intended to convey that similar values ​​promote the equivalent results or effects listed in the claims. That is, it should be understood that amounts, sizes, formulas, parameters and other quantities and characteristics are not precise and do not need to be precise, but can be approximate and / or larger or smaller, reflecting tolerances, conversion factors, rounding, measurement errors, etc., as needed, and other factors known to those skilled in the art. Typically, amounts, sizes, formulas, parameters or other quantities or characteristics are "about" or "approximately", whether or not explicitly stated. It should be understood that, unless explicitly stated otherwise, when "about" is used before a quantitative value, the parameter also includes the specific quantitative value itself.

[0036] Unless otherwise indicated, numerical values ​​should be understood to include the same value when reduced to the same number of significant figures and to include values ​​that differ from the stated value by less than the experimental error of ordinary measurement techniques of the type described in this application for determining the value.

[0037] All ranges disclosed herein include the endpoints listed and are independent of the endpoints. The ranges disclosed herein and the endpoints of any value are not limited to the exact ranges or values. They are not precise enough and may include values ​​that approximate these ranges and / or values.

[0038] As used herein, approximate language may be applied to modify any quantitative representation that can change without causing a change in the basic function to which it is associated. Therefore, in some cases, a value modified by one or more terms such as "about" and "substantially" may not be limited to the specified exact value. In at least some instances, approximate language may correspond to the precision of the instrument used to measure the value. The modifier "about" should also be considered to disclose a range defined by the absolute values ​​of the two end values. For example, the expression "about 2 to about 4" also discloses a range of "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 from 0.9-1.1. Other meanings of "about" can be understood from the context, such as rounding, so for example, "about 1" can also mean from 0.5 to 1.4. In addition, the term "comprising" should be understood to have the open meaning of "including", but the term also includes the closed meaning of the term "consisting of..." For example, a composition comprising component A and component B may be a composition comprising A, B, and other components, but may also be a composition made only of A and B. Any document cited herein is incorporated by reference in its entirety for any and all purposes.

[0039] Now refer to Figure 1 According to the present disclosure, an incubator system 100 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 adjust the local environment contained within the sample chamber 106. As shown in the figure, the control unit 102 can be connected to at least one inlet 107 of the sample chamber 106 via at least one connector 104.

[0040] Now refer to Figure 2The control unit 102 may include a gas mixing manifold 110 that allows for the mixing of gases received from one or more sources. In some embodiments, the one or more sources may include one or more external sources (i.e., located outside the manifold 110), such as one or more gas tanks, gas generators, or other external gas supplies. In some embodiments, the one or more sources may include one or more sources located within the manifold 110. The manifold 110 may include multiple gas regulating valves and may be connected to external gas supplies, which may be mixed within the manifold to achieve the desired experimental conditions. Gas supplies may include, but are not limited to, nitrogen (N2), carbon dioxide (CO2), oxygen (O2), air, and the like, including one or more combinations thereof. Valves connected to the external gas supplies may be actuated to allow and block the entry of external gases into the manifold 110. The manifold 110 may also include one or more sensors configured to detect the levels of one or more gases. Thus, a user may connect the manifold 110 to the desired gases and then actuate the valves to blend the gases to achieve a mixture within the manifold 110 that meets the desired experimental conditions. For example, a user may set the O2 level to a value within the range of approximately 0% to approximately 21%, including any and all intermediate values. Similarly, a user may set the CO2 level to a value within the range of approximately 0% to approximately 20%, including any and all intermediate values.

[0041] The composition of the gas mixture within manifold 110 can change over time, i.e., from a first blend at a first time to a second blend at a second time. This, in turn, allows a user to expose a sample in fluid communication with gas manifold 110 to different gas conditions at different times. A pump 108 (such as air pump 108) can be used to draw air from outside the control unit and / or draw gas from the gas mixing manifold 110 within the control unit. Control unit 102 can include a water reservoir 114 that can be located within the control unit. In some examples, the lower portion of the reservoir can be made of aluminum or another thermally conductive material, and the upper portion can be made of polycarbonate or other plastic. By way of non-limiting example, the lower portion of the reservoir can be made of a material that effectively conducts heat, and the upper portion of the reservoir can be transparent to allow a user to see the water level therein. A heater, such as heater plate 116, can be used to heat water reservoir 114, which in turn can produce humidified air. By way of non-limiting example, heater plate 116 can be a 60-watt (W) heater and can include a digital temperature sensor. Conditioned air, which may include at least some of the contents of the gas mixing manifold 110 and / or heated humidified air from the water reservoir 114, may be supplied via the connector 104 (see Figure 1) is delivered from the control unit 102 to the sample chamber 106. For example, the humidity in the sample chamber 106 is controlled by one or more valves on the gas mixing manifold 110. A valve can be actuated to allow the gas mixture to flow through the water reservoir 114, thereby humidifying the gas mixture. If lower humidity is preferred, the valve is at least partially closed, and a second valve is opened, 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 the user to deliver air at a desired temperature to the sample chamber 106. The pump 108 may be located within the control unit 102 and can deliver conditioned (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, thereby allowing the user to perform longer experiments while maintaining sample viability and more accurate sample medium volume levels in the absence of evaporation.

[0042] like Figure 2 As shown, by way of non-limiting example, the control unit 102 may include one or more of various sensors, such as an oxygen sensor 118 and a carbon dioxide (CO2) sensor 112. The control unit 102 may also include one or more gas filters / regulators 120. The gas filters / regulators 120 may be used, for example, to condition the gas delivered from an external gas source to the gas mixing manifold 110 and remove particles from the circulating air through filtration. In some embodiments, the filters / regulators remove particles larger than approximately 5 µm from the circulating air. The filters / regulators operate to control the pressure of the gas reaching the manifold 110. In some embodiments, the circulating gas is regulated to a pressure of approximately 35 psi. The control unit 102 may 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 may be mounted on or adjacent to the manifold 110, or in fluid communication with the manifold 110.

[0043] Figure 3 An external view is provided showing 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 is operative to filter air from the air source. Figure 2The air pump 108 shown is used to filter the air drawn in. The control unit 102 may include one or more gas inlets 126b that may be connected to an external gas supply source, such as a supply source of O2, N2, CO2 gas and / or air. The control unit 102 may also include a power supply connection 126c, a power supply connection 126d to the sample chamber 106 (see Figure 1 ) data and / or power connection 126d, and a data port 126e (eg, a USB port) that allows connection between the control unit 102 and an external computer or other control device.

[0044] Now refer to Figure 4A , shows a partially exploded top perspective view of a sample chamber 106 according to an exemplary embodiment. As shown, the sample chamber 106 may include a chamber housing 150 that defines an interior volume 135 for holding one or more sample containers 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 lid or "cover" 122 for coupling with the chamber housing 150 and enclosing a portion of the interior volume 135. The cover 122 preferably includes a window 124 for viewing the sample media located in the interior volume 135 of the sample chamber 106. The cover 122 will be discussed in more detail below. The chamber housing 150 includes at least one inlet 107 that is accessible via at least one connector 104 (see Figure 1 ) receives conditioned air from the control unit 102 to deliver it to the interior volume 135.

[0045] The chamber housing 150 has a first face 152 and a second face 154 that is opposite the first face 152 along a first direction Z. The first face 152 is configured to face an imaging lens, such as the objective lens 142, along the first direction Z. The second face 154 is configured to be mounted together with the lid 122. The lid 122 can be fixedly mounted to the second face 154. Alternatively, the lid 122 can be a peel-off lid that is not otherwise fixed to the chamber housing 150.

[0046] The first face 152 is spaced apart from the second face 154 of the chamber housing 150 along a first direction Z in a lens-facing direction Z1, while the second face 154 is spaced apart from the first face 152 in a lens-away direction Z2, which is opposite the lens-facing direction Z1. It should be understood that the lens-facing direction Z1 and the lens-away direction Z2 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 oppose each other along a second direction X that is substantially perpendicular to the first direction Z. The chamber housing 150 also includes a first side wall 160 and a second side wall 162 that oppose each other along a third direction Y that is substantially perpendicular to the first and second directions. The first and second end walls 156 and 158, as well as the first and second side walls 160 and 162, substantially enclose the interior volume 135 relative to the second and third directions X and Y. The interior volume 135 is configured to contain a local environment therein, preferably an incubation local environment for receiving one or more sample containers. The sample container 106 is configured such that a user can remove the lid 122 from the chamber housing 150 , place one or more sample containers within the interior volume 135 , and replace the lid 122 atop the second face 154 , thereby enclosing the interior volume 135 relative to the away-from-lens direction Z2 .

[0047] In the illustrated embodiment, sample chamber 106 is configured to be placed atop a stage (e.g., a movable x-stage, y-stage) of a microscopy imaging system. Thus, during use, when sample chamber 106 is positioned in this orientation, first direction Z is vertical, and second direction X and third direction Y are each horizontal. In such an embodiment, direction Z1 toward the lens can be characterized as "downward" direction Z1, and direction Z2 away from the lens can be characterized as "upward" direction Z2. It should be understood that, as used herein with reference to the illustrated embodiment (e.g., when referring to the spatial relationships between various features), directional terminology can be used to indicate the spatial relationships between various features of sample chamber 106. For example, the terms "downward," "lower," "beneath," "bottom," "under," and their derivatives refer to the downward direction Z1; and the terms "upward," "upper," "above," "top," "atop," and their derivatives refer to the upward direction Z2. By way of some specific, non-limiting examples, when referring to the embodiments illustrated herein, the first face 152 of the housing body 150 may also be referred to as the "lower" face 152; and the second face 154 of the housing body 150 may also be referred to as the "upper" face 154. Similar such directional terms are also used herein to describe other features of the illustrated embodiments. However, in other embodiments, the sample chamber 106 may be adapted such that, during use, the first direction Z is offset relative to the vertical direction (and, by extension, one or both of the second and third directions may be offset relative to the horizontal direction). It should be understood that, unless otherwise indicated herein, the aforementioned 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 relative to the vertical direction. By way of two such examples, the sample chamber 106 may be adapted for use with a microscopy imaging system in which the imaging lens faces downward rather than upward, or alternatively faces horizontally rather than vertically. The reader will understand that in such alternative configurations, the "bottom face" 152 of the chamber housing 150 faces the imaging lens even if the lower face 152 is positioned above the upper face 154 (in the case of a downwardly facing lens), or even if the lower face 152 and upper face 154 are horizontally spaced apart from each other rather than vertically (in the case of a horizontally facing lens). Summarizing in a different way, the directional terms used herein indicate spatial relationships between various features, and unless otherwise indicated herein, those spatial relationships will apply regardless of the particular orientation in which the sample chamber 106 is oriented in three-dimensional space.

[0048] Now refer to Figure 4B, chamber housing 150 can include a first or "lower" housing body 150a and a second or "upper" housing body 150b that can be connected to each other. In the illustrated embodiment, lower housing body 150a defines a lower face 152, upper housing body 150b defines an upper face 154, and lower housing body 150a and upper housing body 150b can be connected to each other in a sandwich-like manner. As shown, the lower housing body 150a and the upper housing body 150b can each include respective first and second body end walls 156a, 156b, 158a, 158b and first and second body side walls 160a, 160b, 162a, 162b, which, when coupled together, form portions of the first and second end walls 156, 158 and the first and second side walls 160, 160 of the chamber housing 150. The lower housing body 150a and the upper housing body 150b also preferably each define a central opening 155a, 155b extending therethrough along a first direction Z. The central opening 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 a sample medium placed in the sample chamber 106. The central opening 155b of the upper housing body 150b provides an opening through which a user may place one or more sample containers into the interior volume 135 of the sample chamber 106 when the cover 122 is removed.

[0049] Lower housing portion 150a preferably also includes an interior support surface 153, which may extend in a rim-like manner around the inner perimeter of end walls 156a, 158a and side walls 160a, 162a. Interior support surface 157 may be configured to support various features of sample chamber 106, as described in more detail below. Lower housing portion 150a also preferably includes a platform surface 157, which may be located on a side of first end wall 156a opposite central opening 155a along second direction X. Platform surface 157 may be configured to support various features of chamber housing 150, such as circuitry (e.g., one or more printed circuit boards (PCBs)) and air handling components, as described in more detail below. Upper housing portion 150b preferably includes a cover portion 159, which may cover at least a portion of platform surface 157 and may be configured to cover some or all of the various features supported by platform surface 157.

[0050] The chamber housing 150 may further include a support member 150c disposed between the lower housing body 150a and the upper housing body 150b. The support member 150c has a top end 164 and a bottom end 166 spaced apart from each other along a first direction Z. The support member 150c also includes first and second member end walls 156c, 158d and first and second member side walls 160c, 162c. When the support member 150c is coupled to the lower housing body 150a and the upper housing body 150b, the first and second member end walls and the first and second member side walls form portions of the first and second end walls 156, 158 and the first and second side walls 160, 160 of the chamber housing 150. Inner surfaces 170 of the member end walls 156c, 158c and the member side walls 160c, 162c define respective portions of the interior volume 135. The first member end wall 156c can define an opening 172 for allowing one or more vents 128 to enter the interior volume 135, as described in more detail below. The support member 150c also includes a sample support surface 168 vertically located between the top end 164 and the bottom end 166 and facing upward (i.e., in the upward direction Z2). The sample support surface 168 is configured to hold one or more sample containers 175 (see FIG. 1 ) placed in the interior volume 135 of the sample chamber 106. Figure 5F ). As shown, the sample support surface 168 can extend around the entire inner periphery of the support member 150c in a rim-like manner and be spaced inwardly from the member end walls 156c, 158c and the member side walls 160c, 162c. The support member 150c defines a central opening 155c extending therethrough along the first direction Z. Similar to the lower housing body 150a, the central opening 155c of the support member provides an open, unobstructed space between the sample chamber 106 and the imaging lens. In addition, similar to the upper housing body 150b, the central opening 155c of the support member 150c also provides an upper opening through which a user places one or more sample containers into the interior volume 135 and atop the sample support surface 168. The support member 150c can be made of a material that provides advantageous thermal conductivity and insulation for incubating the interior volume 135, including, by way of non-limiting example, aluminum, steel, and titanium. In such embodiments, the standoff member 150c may also be referred to as a "heat sink" 150c. It should be understood that the standoff member 150c may alternatively be made from virtually any material that is capable of being machined, physically stable, and thermally conductive.

[0051] The sample chamber 106 includes at least one inlet 107 that is communicable with a chamber manifold 130 that is 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 further defines internal vents 128 at an opposite end thereof. The internal vents 128 are configured to extend through or at least reside within openings 172 in the first end wall 156c of the support member 150c and to deliver air received from the inlet 107 to the interior volume 135 of the sample chamber 106.

[0052] Continue to refer Figure 4B , the sample chamber 106 includes various thermal regulation features that can be used to heat the environment within the interior volume 135 to a desired temperature, such as for incubating a sample medium placed therein. For example, the thermal regulation features can be configured to heat the environment within the interior volume 135 to a temperature within a range of about 30°C to about 40°C, and more specifically to a temperature within a range of about 35°C to about 40°C, and more preferably to about 37°C. The thermal regulation features can also be used to maintain and / or adjust the temperature within the interior volume 135 as desired. Examples of such thermal regulation features will now be described. For example, the sample chamber 106 can include a first heater 138 that can extend around the periphery of the interior volume 135. The first heater 138 can have a base heater portion 144 that extends around the periphery of the interior volume 135 and defines a central opening 155d. The base heater portion 144 can be configured to be positioned below and heat the bottom end 166 of the support member 150c as needed to maintain a desired temperature within the interior volume 135. The first heater 138 preferably also has one or more wall heater portions 146 that extend upwardly along corresponding side walls or end walls of the chamber housing 150. In the illustrated embodiment, the first heater 138 includes a pair of wall heater portions 146 that extend upwardly from the base portion 138 along the first side walls 160a to 160c and the second side walls 162a to 162c. The pair of wall heater portions 146 can be configured to heat the side walls 160c, 162c of the support member to further adjust the temperature of the support member 150c as needed to maintain a desired temperature within the interior volume 135.

[0053] The sample chamber 106 may also include an inlet heater 148 for heating air delivered through the chamber manifold 130. The inlet heater 148 may include a lower panel 174 and an upper panel 176 that contact upper and lower portions of the chamber manifold 130. In this manner, the inlet heater 148 may operate in conjunction 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 may also include circuitry features (such as flex circuits, tracers, connecting pins, headers, etc.) for electrically connecting the heaters 138, 148 to the control unit 102, which may include a processor executing computer-readable instructions stored in computer memory for controlling the operation of the heaters 138, 148, among other things. Alternatively, the heaters 138, 148 may be electrically connected to a separate electronic control unit, which may be located within the sample chamber 106.

[0054] It should be understood that, by way of non-limiting example, the cover 122 (see Figure 4A ) may also include one or more thermal regulator features, such as a thermal insulation film and / or a conductive heater. The thermal insulation film and / or the conductive heater may be superimposed on a portion or all of the window 124 of the cover 122. Therefore, the window 124 may also be characterized as a heater or a thermal insulator. The conductive heater and / or the thermal insulator of the cover 122 may be used alone or in combination with the first heater 138 and / or the inlet heater 148 to heat the environment within the sample chamber 106 to a desired temperature. By way of non-limiting example, the thermal insulation film of the cover 122 may include one or more suitable thermal insulation 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 internal volume 135 to the control unit 102, as described in more detail below.

[0055] Continue to refer Figure 4BSample chamber 106 includes an air circulator or "actuator" unit 132 for directing air to a target location located between the interior volume 135 of sample chamber 106 and the imaging lens relative to a first direction Z. This target location is preferably located beside and below a lower face 152 of housing body 150, which may be referred to as a location "below" sample chamber 106. Preferably, air actuator unit 132 is configured to direct conditioned air (i.e., air having a regulated (e.g., heated) temperature) to the target location between interior volume 135 and the imaging lens. By heating this air and directing it to the target location, sample chamber 106 can effectively heat or warm the imaging lens in a manner that inhibits or at least reduces 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 out or remove any existing condensation that may have accumulated on the imaging lens, thereby improving image quality. The air actuator unit 132 includes an air actuator 180, which can be a fan 180 or other mechanism for drawing in air (such as ambient air adjacent to the sample chamber 106) and directing the drawn-in air through the air actuator unit 132. The air actuator unit 132 preferably also includes a duct member 182 that directs the air drawn in from the air actuator 180 along one or more channels 184 to one or more outlet vents or ports 140 facing a target location. The air actuator unit 132 can include an air actuator filter 134 for filtering the air drawn into the duct member 182 by the air actuator 180. As shown, the air actuator filter 134 can be carried by a filter support 137 that can be coupled to and detached from the duct member 182. The air actuation unit 132 can reside in a compartment 188 defined by portions of the lower housing body 150a and the upper housing body 150b, such as along the first sidewalls 160a, 160b thereof, as in the illustrated embodiment. It should be understood that the air actuation unit 132 can be removable and / or replaceable from the sample chamber 106, as described in more detail below.

[0056] Now refer to Figure 5A, shows a partial view of the sample chamber 106, wherein the upper housing body 150b has been removed for visualization purposes to show the air actuator unit 132. As shown, the air actuator 180 can draw or otherwise introduce air (e.g., external ambient air, as indicated by arrows 145) into the conduit member 182, particularly through an opening 181 defined in an exterior face 183 of the conduit member 182. The housing 150 preferably defines a vent port 190 adjacent to the air actuator 180 to allow external air to enter the air actuator unit 132. Figure 5B As shown, once drawn into conduit member 182 via air actuator 180, the introduced air is directed through conduit member 182 as indicated by the arrows, directed toward one or more outlet ports 140, which are preferably located below the sample container present within the interior volume 135 of sample chamber 106, as described in more detail below. Figure 5C As shown, the air actuator filter 134 and the filter support 137 can reside within a filter receiver 192 defined by the chamber housing 150 (such as defined by both the lower housing body 150a and the upper housing body 150b of the chamber housing). The filter receiver 192 and the filter support 137 can be configured to facilitate filter replacement by pulling the filter support 137 upward from the receiver 192, as shown.

[0057] Now refer to Figures 5D to 5E , shows an interior face 185 of the duct member 182 that faces the interior volume 135 of the sample chamber 106. The duct member 182 defines one or more channels 184 that direct the incoming air flow to one or more outlet vents or ports 140. In the illustrated embodiment, the duct member 182 defines a single channel 184 that extends therethrough and defines a flow path to the one or more outlet ports 140. As shown, the air actuation unit 132 may include an outlet member 194 that is connectable adjacent to the duct member 182, particularly adjacent to the underside 196 of the duct member 182. The channel 184 may be characterized as having multiple portions, such as: a first or open portion 184a that may be aligned with the air actuator 180 along a third direction Y; a second or main portion 184b that may extend along a second direction X and alongside one of the wall heater portions 146 of the first heater 138 (see Figure 4B ); and a third or redirecting portion 184c that redirects the incoming air flow behind and below the lower face 152 of the housing body 150. For illustrative purposes, Figure 5ESome exemplary air flow paths are shown in FIG. The second main portion 184b of the channel 184 can be defined between the interior face 198 of the inner wall 200 of the duct member 182 and the associated wall heater portion 146 (see FIG. Figure 4B ). The associated wall heater portion 146 can be positioned adjacent to or in contact with the interior face 185 of the duct member 182. In this way, the air introduced into the channel 184 is guided along the second direction X beside the associated wall heater portion 146, which can heat the introduced air to a temperature sufficient to inhibit or at least reduce the formation of condensate at the imaging lens. At the redirecting portion 184c of the channel 184, the introduced air is guided generally in the third direction Y around the bend located downstream of the end surface 202 of the inner wall 200. Within the redirecting portion 184c downstream of the bend, the introduced air is further redirected downward (direction Z1) through the interior face 204 of the outer wall 206 of the duct member 182. The downwardly redirected air passes through the bottom opening 208 of the duct member 182 (at Figure 5E ) and the associated bottom opening 210 of the lower housing body 150a (shown in Figure 5F ), and enters an outlet passage 212 at least partially defined by outlet member 194.

[0058] Now refer to Figure 5FThe outlet passage 212 is partially defined by the outlet base surface 214 of the outlet member 194 and may be further partially defined by the lower face 152 of the lower housing body 150a. In the illustrated embodiment, the outlet passage 212 is vertically defined between the outlet base surface 214 of the outlet member 194 and the lower face 152 of the lower housing body 150a. As shown, the outlet base surface 214 has a curved portion that redirects the incoming air passing through the bottom openings 208, 210 generally along the third direction Y. The outlet base surface 214 also has a flat portion that guides the incoming air along the outlet passage 212 in the third direction Y and toward a target location below the lower face 152 of the chamber housing 150. As shown, the flat portion of the outlet base surface 214 is preferably substantially parallel to the lower face 152 of the chamber housing 150. This causes the introduced air to exit the outlet member 194 in respective air flow directions that are substantially parallel to the lower face 152. Additionally, the outlet member 194 preferably includes guide members (such as baffles or fins 216) that project within the outlet passage 212 and are configured to diffuse the introduced air exiting the outlet port 140 in respective directions having a directional component along the second direction X, thereby expanding the target location to extend alongside a larger portion of the lower face 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 advantageously enhancing the suppression (or at least reduction) of condensate formation on the imaging lens. Figure 5G As shown, the fins 216 are elongated along respective directions D1 to D4 that diverge from one another in the downstream direction of the airflow D0, thereby causing the introduced air to diffuse outwardly when leaving the outlet port 140. Figure 5F As shown, the fins 216 may extend upwardly and contact or at least be proximate to the lower face 152 of the chamber housing 150 , thereby providing a plurality of outlet ports 140 along the second direction X between the fins 216 .

[0059] Now refer to Figure 6A and Figure 6B , illustrate exemplary embodiments of an air actuator 180 for use with the air actuator unit 132. In these embodiments, the air actuator 180 includes a fan 180 having a front face 230 ( Figure 6A The fan 180 also has a back surface 232 ( Figure 6B), which back side is configured to be adjacent to the interior face 198 of the inner wall 200 of the conduit member 182. It will be appreciated that the fan 180 provides numerous benefits for the air actuation unit 132. As the inventors have discovered, one benefit is that, when used with sample containers of various sizes and configurations, the fan 180 effectively inhibits or at least reduces condensation accumulation and / or fogging of the objective lens. Another such benefit is that the fan 180 also helps maintain the sample container (and the sample medium therein) at a uniform temperature, thereby enhancing the incubation environment for the sample medium within the sample chamber 106.

[0060] Thus, according to the above-described embodiments, air (such as ambient air) can be introduced into the air actuator unit 132 and heated by the heater 138, and the heated air can then be directed out of one or more outlet vents or ports 140 and against, along, or near an imaging lens (e.g., objective lens 142) present below the sample chamber 106. This heated air can, in turn, inhibit or at least reduce fogging and / or condensation accumulation 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 a region between the imaging lens and the sample chamber 106.

[0061] Now refer to Figure 7 The sample chamber 106 preferably includes at least one sensor, such as a temperature / humidity sensor 136, which monitors the environment of a target location 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 air actuator unit 132, the heaters 138, 148, and / or other features of the sample chamber 106 disclosed herein. In this way, the disclosed incubator 100 can (1) generate a locally modulated environment within the interior of the sample chamber 106, wherein the locally modulated environment includes, for example, a specified temperature, humidity, and (2) reduce or eliminate fogging of an imaging lens (such as the objective lens 142) that images one or more samples disposed within the sample chamber 106.

[0062] Now refer to Figure 8A and Figure 8B , which show photographs of the objective lens 142 having condensation thereon due to a temperature difference between the objective lens 142 and a sample container of the prior art. As shown in these photographs, a large amount of condensation has accumulated on the objective lens 142, which may significantly obscure the image that can be obtained by the objective lens 142 and may require time and effort to remove based on prior art techniques.

[0063] Now refer to Figure 9, showing a process flow according to an exemplary method of the present disclosure. As shown in the figure, at step 1, the user can set a first set of desired conditions for the interior 135 of the sample chamber 106, for example, the temperature, humidity, and gas level to which the sample in the sample chamber 106 will be exposed. The user can then, for example, automatically actuate the gas manifold 110, the pump 108, and other features of the disclosed incubator 100 at step 2 to achieve a set of desired conditions. Then, at step 3, the user can image the sample located in the interior 135 of the sample chamber 106 after being exposed to the set of conditions for a desired period of time. Then, at step 4, the user can set the next set of desired conditions, such as the environmental conditions inside the sample chamber 106. For example, the environmental conditions include the specified temperature, humidity, and / or gas mixture level or gas mixture composition to which the sample in the sample chamber 106 is exposed, and then perform the aforementioned steps as needed.

[0064] Now refer to Figure 10 , a schematic diagram depicting an exemplary air flow provided by an air actuation unit 132 according to another embodiment of the present disclosure is shown. Specifically, in this embodiment, ambient air 99a is introduced into and passes through the air actuation unit 132, and is directed out of one or more outlet ports 140 as exhaust air 99b in a direction intersecting an objective lens 142 of a microscope located below the sample chamber 106. As in the above-described embodiments, the introduced ambient air 99a of this embodiment may be heated by a heater 138 (see FIG. 1 ) as the air travels through the air actuation unit 132. Figure 4B ) is heated and can then be directed as exhaust air 99b toward, through, or adjacent to the objective lens 142 to dry or remove any condensation that may have accumulated and / or prevent any condensation from accumulating on the objective lens 142, thereby improving image quality.

[0065] Advantages of the disclosed sample chamber 106 design include improved heating uniformity of incubated samples located therein. Table 1 provides test data showing an array of temperature measurements taken at various locations within a 96-well sample plate incubated within the sample chamber 106 described herein.

[0066] Table 1 - Array of temperature measurements from individual wells of a sample plate incubated in a chamber contemplated by the present disclosure .

[0067] The test results shown in Table 1 demonstrate that relatively uniform heating was achieved and maintained within the sample plate incubated within the sample chamber 106 of the present disclosure. The highest temperature measured in 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 with the sample chamber 106 of the present disclosure provides improvements in experimental design and experimental results obtained from samples contained within the sample chamber 106 disclosed herein.

[0068] It should be understood that the sample chamber 106 can be adapted to receive sample containers of various sizes and shapes as desired.

[0069] It should also be understood that in additional embodiments, the incubator system 100 may 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, and these sample containers are interchangeable with each other in the sample chamber 106.

[0070] It should also be understood that in additional embodiments, the air actuation unit can use compressed air as an alternative to fan 180 to inhibit or at least reduce the accumulation of condensation on the imaging lens and / or fogging. In such embodiments, the air actuation unit can include a reservoir of compressed air and a nozzle through which the compressed air can be released and directed toward, through, or adjacent to the imaging lens. In this way, the compressed air can create a pressure differential at a target location between the sample chamber and the imaging lens. This pressure differential can be used to lower the dew point temperature at the target location, thereby inhibiting or at least reducing the accumulation of condensation on the imaging lens and / or fogging of the imaging lens.

[0071] It should be understood that according to further additional embodiments herein, the air actuation unit 132 may be considered to include means for adjusting air conditions at a target location on, at, or adjacent to the imaging lens. In such embodiments, the means for adjusting air conditions in the target location may include a fan 180, compressed air, or other features and techniques. The air actuation unit 132 according to such embodiments may also include means for directing conditioned air to the target location, which may include a duct member, such as the duct member 182 described above, and may also include an outlet member, such as the outlet member 194 described above. The means for directing conditioned air to the target location may also include fins or baffles, such as the fins 216 described above.

[0072] It should also be understood that when numerical prepositions (e.g., "first," "second," "third") are used herein with reference to an element, component, dimension, or feature thereof (e.g., a "first" sensor, a "second" sensor), such numerical prepositions are used to distinguish the element, component, dimension, and / or feature from another such element, component, dimension, and / or feature, and are not limited to the specific numerical prepositions 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, as long as the element, component, dimension, and / or feature remains appropriately distinguished in the context in which the numerical prepositions are used.

[0073] Although the present disclosure has been described in detail, it should be understood that various changes, substitutions and modifications may be made without departing from the spirit and scope of the present invention as defined by the appended claims. In addition, the scope of the present disclosure is not intended to be limited to the specific embodiments described in the specification. Specifically, one or more features from the features of the aforementioned embodiments may be adopted in other embodiments herein. As will be readily appreciated by those of ordinary skill in the art, according to the present disclosure, currently existing or later to be developed processes, machines, products, material compositions, components, methods or steps that perform substantially the same functions as the corresponding embodiments described herein or achieve substantially the same results as the corresponding embodiments may be utilized.

[0074] aspect The following aspects are merely illustrative and do not limit the scope of the present disclosure or the appended claims.Any one or more portions of any one or more aspects may be combined with any one or more portions of any one or more other aspects.

[0075] Aspect 1. A system for improved imaging, the system comprising: a sample chamber configured to contain the local environment therein; and Air actuator, The sample chamber is configured such that air excited by the actuator is directed between the sample chamber and an objective lens configured for imaging a sample within the sample chamber, the air being directed so as to reduce or eliminate condensation on the objective lens.

[0076] Aspect 2. The system of aspect 1, wherein the sample chamber includes features for supporting a sample container disposed within the sample chamber.

[0077] Aspect 3. The system of aspect 1, wherein the sample chamber comprises a lower region for engaging with a sample container.

[0078] Aspect 4. The system of aspect 3, wherein the lower zone comprises a heating element.

[0079] Aspect 5. The system of aspect 4, wherein the heating element is configured to heat the sample chamber to approximately 37°C.

[0080] Aspect 6. The system of aspect 4, wherein the heating element further heats the air excited by the air actuator.

[0081] Aspect 7. The system of aspect 1, wherein the sample chamber comprises a lid.

[0082] Aspect 8. The system of aspect 7, wherein the cover comprises a heating element.

[0083] Aspect 9. The system of aspect 8, wherein the heating element is configured to heat the sample chamber to 37°C.

[0084] Aspect 10. The system of aspect 1, wherein the chamber comprises a manifold and an outlet, wherein the manifold is configured to direct air energized by the actuator to the outlet.

[0085] Aspect 11. The system of aspect 10, wherein the outlet is aligned with the objective lens.

[0086] Aspect 12. The system of aspect 11, wherein the outlet is movable.

[0087] Aspect 13. The system of aspect 12, wherein the outlet is slidable, rotatable, or both.

[0088] Aspect 14. The system of aspect 10, wherein the manifold further comprises one or more sensors.

[0089] Aspect 15. The system of aspect 14, wherein the one or more sensors include a temperature sensor.

[0090] Aspect 16. The system according to aspect 1, further comprising a control unit that delivers conditioned air to the sample chamber.

[0091] Aspect 17. The system of aspect 16, wherein the control unit comprises a gas mixing manifold.

[0092] Aspect 18. The system according to 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 selected concentration mixture.

[0093] Aspect 19. The system according to aspect 16, wherein the control unit comprises a pump configured to excite the conditioned air to the fluid inlet of the sample chamber.

[0094] Aspect 20. The system of aspect 19, wherein the pump is positioned inside the control unit.

[0095] Aspect 21. The system of aspect 16, wherein the conditioned air comprises humidified air or heated humidified air.

[0096] Aspect 22. The system of aspect 16, wherein the conditioned air comprises dry air or heated dry air.

[0097] Aspect 23. The system of aspect 21, wherein the humidified air has a humidity of about 50% to about 90% humidity.

[0098] Aspect 24. The system of aspect 18, wherein the selected concentration mixture comprises about 5% to about 12% carbon dioxide.

[0099] Aspect 25. The system of aspect 18, wherein the selected concentration mixture comprises at most about 21% oxygen.

[0100] Aspect 26. The system of aspect 18, wherein the selected concentration mixture comprises about 67% to about 95% nitrogen.

[0101] Aspect 27. A system according to any one of Aspects 1 to 26, wherein the system is configured to operate so that the lowest temperature of all positions of the sample container located within the sample chamber is within about 15% of the highest temperature of all positions of the sample container.

[0102] Aspect 28. A method comprising: operating a system according to any one of Aspects 1 to 27 to (1) reduce or eliminate condensation on the objective lens, (2) maintain a first environment within a sample chamber that is different from the ambient environment outside the sample chamber in one or more of temperature, humidity, and gas mixture, or both (1) and (2).

[0103] Aspect 29. A method according to Aspect 28, further comprising operating a system according to any one of Aspects 1 to 27 so as to maintain a second environment within the sample chamber, the second environment being different from the first environment in one or more of temperature, humidity, and gas mixture relative to the ambient environment outside the sample chamber.

[0104] Aspect 30. The method according to any one of aspects 28 to 29, wherein the operation is performed so that during the operation, the lowest temperature of all positions of the sample container located within the sample chamber is within about 15% of the highest temperature of all positions of the sample container.

[0105] Aspect 31. An incubator comprising: a sample chamber comprising an interior and (i) a fluid inlet, (ii) a surface for receiving a sample specimen container, and (iii) one or more heating elements; and a control unit configured to engage with 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 the interior of the sample chamber.

[0106] The sample chamber can be placed on a microscope stage and / or a microscope slide. A gasket can be used to seal the sample chamber to the microscope stage and / or slide.

[0107] Aspect 32. The incubator of Aspect 31, wherein the control unit comprises a gas mixing chamber, which in some instances is also referred to as a gas mixing manifold. The gas mixing chamber may comprise any one or more of a valve, a sensor, a filter, and / or a regulator for generating a gas mixture for delivery to the fluid inlet. One or more sensors may be positioned to monitor any one or more of the gas content, temperature, and humidity of the contents of the gas mixing manifold.

[0108] Aspect 33. The incubator of Aspect 32, wherein the gas mixture within the gas mixing chamber comprises one or more gases selected from oxygen, carbon dioxide, nitrogen, and standard air, the gases being balanced to a selected concentration mixture. One or more of these gases may be received from a gas source (e.g., a gas tank) external to the gas mixing chamber. By regulating the introduction of different gases into the gas mixing chamber, a user may achieve a mixture within the gas mixing chamber that meets set specifications for gas levels.

[0109] Aspect 34. The incubator of Aspect 31, wherein the conditioned air comprises humidified air or heated humidified air. For example, a user may set a desired humidity level, which may be achieved by humidifying the air drawn into the control unit, by mixing humidified air formed within the control unit with air drawn into the control unit, or both. It will be appreciated that the contents of the gas mixing manifold may be mixed with humidified air and / or non-humidified air.

[0110] Aspect 35. The incubator according to aspect 31, wherein the conditioned air comprises dry air or heated dry air. The incubator may include a dehumidifier, for example, a dehumidifier configured to dehumidify the air delivered to the inlet of the sample chamber via a connector.

[0111] Aspect 36. The incubator according to aspect 31, further comprising a lid.

[0112] Aspect 37. The incubator of Aspect 36, wherein the lid comprises (i) a thermally insulating film coating, (ii) a conductive heater, or both. The lid can be connected to a power source, which in turn is used to heat the lid. The lid can also include a power source, such as a battery, for powering the lid heater.

[0113] Aspect 38. The incubator of aspect 31, wherein the control unit comprises a pump configured to excite the conditioned air into the fluid inlet.

[0114] Aspect 39. The incubator of aspect 38, wherein the pump is positioned inside the control unit.

[0115] Aspect 40. The incubator according to aspect 34, wherein the humidified air is humidified to a humidity of about 50% to about 90%, preferably to a humidity of about 80%. Humidity levels of 50% to 90%, 55% to 85%, 60% to 80%, or even 70% are all suitable.

[0116] Aspect 41. The incubator of aspect 33, wherein the selected mixture comprises about 5% to about 12% carbon dioxide, preferably about 5% carbon dioxide. Carbon dioxide levels of about 5% to about 12%, about 6% to about 11%, about 7% to about 10%, or even about 8% to about 9% are all considered suitable.

[0117] Aspect 42. The incubator of aspect 33, wherein the selected mixture comprises up to about 21% oxygen. The mixture may have less than 21% oxygen, for example, about 0.5% to about 20%, about 1% to about 19%, about 2% to about 18%, about 3% to about 17%, about 4% to about 16%, about 5% to about 15%, about 6% to about 14%, about 7% to about 13%, about 8% to about 12%, about 9% to about 11%, or even about 10% oxygen.

[0118] Aspect 43. The incubator of aspect 33, wherein the selected mixture comprises about 67% to about 95% nitrogen, preferably about 75% nitrogen. The mixture may comprise 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.

[0119] Aspect 44. The incubator according to aspect 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.

[0120] Aspect 35. The incubator of aspect 31, wherein the one or more heating elements are configured to heat the sample specimen container to approximately 37°C.

[0121] Aspect 46. The incubator of aspect 31, further comprising a manifold configured to receive the conditioned air.

[0122] Aspect 47. The incubator of aspect 36, wherein the manifold is configured to distribute the conditioned air within the interior of the sample chamber. The manifold may extend around a portion of the perimeter (inner perimeter or outer perimeter) of the interior of the sample chamber, or even around the entire perimeter (inner perimeter or outer perimeter) of the interior of the sample chamber.

[0123] Aspect 48. The incubator according to any one of aspects 46 to 47, further comprising at least one vent in fluid communication with the manifold, the at least one vent being configured to direct conditioned air received from the control unit to the sample container. The conditioned air directed to the sample container may be heated by the one or more heating elements of the sample chamber.

[0124] Aspect 49. The incubator according to aspect 31, further comprising an air circulator embodied in the sample chamber, the air circulator being configured to energize air below the sample chamber. Such air can be used to reduce or even eliminate fogging of an objective lens located below the sample chamber.

[0125] Aspect 50. The incubator of aspect 49, wherein the air actuator comprises a fan.

[0126] Aspect 51. The incubator according to any one of aspects 31 to 50, further comprising at least one sensor configured to detect humidity, gas levels, or both.

[0127] Aspect 52. The incubator according to 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 embodied within the control unit, the water reservoir being in fluid communication with the fluid inlet.

[0128] Aspect 53. The incubator of any one of aspects 31 to 53, wherein the incubator is configured to include (i) the gas mixture, (ii) humidified air, or both (i) and (ii) in the conditioned air.

[0129] Aspect 54. An incubator according to any one of Aspects 31 to 53, wherein the incubator regulates the atmosphere in the interior of the sample chamber to at least one specified level of any one or more conditions selected from the group consisting of: oxygen level, carbon dioxide level, nitrogen level, humidity percentage, or temperature.

[0130] Aspect 55. The incubator according to aspect 31, wherein the gas mixing chamber mixes gas received from outside the incubator.

[0131] Aspect 56. The incubator of Aspect 31, wherein the incubator is configured to be mounted on a microscope stage.

[0132] Aspect 57. An incubator according to Aspect 31, wherein the chamber comprises one or more ports configured for introducing one or more materials into the interior of the sample chamber or removing one or more materials from the interior of the sample chamber.

[0133] Aspect 58. A method for imaging a sample in a controlled environment, the method comprising: positioning a sample in a sample chamber of an incubator according to any one of aspects 31 to 57; and acquiring one or more images of the sample in the sample chamber. Image acquisition can be performed, for example, via an inverted microscope.

[0134] Aspect 59. The method of aspect 58, further comprising exposing the sample positioned within the sample chamber to a first set of environmental conditions in the atmosphere of the interior of the sample chamber for a first time interval.

[0135] Aspect 60. The method of aspect 59, wherein the first set of environmental conditions comprises a first specified level of one or more of: oxygen, carbon dioxide, nitrogen, humidity, temperature, or one or more combinations thereof.

[0136] Aspect 61. The method of aspect 59, wherein the first set of environmental conditions includes conditions that are different from corresponding conditions of the atmosphere outside the sample chamber.

[0137] Aspect 62. The method of any one of aspects 58 to 61, wherein the first time interval is in the range of about 1 minute to 72 hours or longer. For example, intervals of about 1 minute to 72 hours or longer, 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.

[0138] Aspect 63. The method of any one of aspects 58 to 62, further comprising exposing the sample to a second set of environmental conditions in the interior of the sample chamber for a second time interval.

[0139] Aspect 64. The method according to aspect 63, wherein the second set of environmental conditions includes conditions different from corresponding conditions of the first set of environmental conditions, including, for example, temperature, humidity, and / or gas mixture. The second time interval may be different from the first time interval.

[0140] Aspect 65. The method according to any one of aspects 58 to 64, further comprising introducing one or more reagents into the sample chamber.

[0141] Aspect 66. The method according to any one of aspects 58 to 65, further comprising extracting one or more samples from the sample chamber.

Claims

1. A sample chamber for use with a sample imager, the sample chamber comprising: A chamber housing, comprising: a first surface and a second surface, the first surface and the second surface being opposite to each other along a first direction, wherein the first surface is configured to face the imaging lens, the second surface is configured to be mounted with a cover having a window, and the first surface and the second surface are spaced apart in a lens-facing direction along the first direction; a first end wall and a second end wall, the first end wall and the second end wall facing each other along a second direction substantially perpendicular to the first direction; and a first side wall and a second side wall, the first side wall and the second side wall being opposite to each other along a third direction substantially perpendicular to the first direction and the second direction, wherein the first and second end walls and the first and second side walls substantially enclose an interior volume relative to the first and second directions, and the interior volume is configured to contain a local environment therein; and An air actuator unit is configured to direct conditioned air to a target position located beside the first face and spaced apart from the first face in the lens-facing direction for suppressing or at least reducing condensation accumulation on the imaging lens.

2. The sample chamber of claim 1 , wherein the air actuator unit comprises an air actuator, a conduit member, and an outlet port, wherein the air actuator is configured to introduce air from outside the sample chamber and guide the introduced air into a channel defined by the conduit member, wherein the channel is configured to guide the introduced air to the outlet port such that the introduced air exits the outlet port and travels outside and beside the second face of the chamber housing. The sample chamber of claim 2 , wherein the air actuator comprises a fan.

4. A sample chamber according to claim 2 or claim 3, wherein the air actuator unit comprises an outlet member in communication with the conduit, the outlet member at least partially defining an outlet passage terminating at the outlet port. 5 . The sample chamber of claim 4 , wherein the outlet member comprises a fin protruding within the outlet passage and configured to direct the flow of air exiting the outlet port. 6 . The sample chamber of claim 5 , wherein the fins are elongated in respective directions diverging from each other in a downstream direction, thereby causing the introduced air to diffuse outwardly as the introduced air exits the outlet port.

7. A sample chamber according to claim 4 or claim 5, wherein the outlet member has an outlet base surface that at least partially defines the outlet passage, the fin extends from the outlet base surface in a direction opposite to the lens-facing direction, and the outlet base surface is substantially parallel to the first face of the chamber housing, so that the introduced air exits the outlet member in a corresponding air flow direction that is substantially parallel to the first face of the chamber housing.

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

9. The sample chamber according to any one of claims 1 to 8, further comprising a heating element configured to heat the interior 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. The sample chamber of claim 9 or claim 10, wherein at least a portion of the heating element is adjacent to the channel such that the introduced air is directed along the at least a portion of the heating element, wherein the introduced air is heated by the heating element. 12 . The sample chamber according to claim 1 , further comprising at least one temperature sensor configured to measure a temperature at the target location.

13. The sample chamber according to any one of claims 1 to 12, further comprising the lid, wherein the lid is configured to be repeatedly attached to and detached from the second side of the chamber housing, and the lid comprises at least one of a heating element or an insulating layer.

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

15. A system for imaging a sample, the system comprising: a control unit for delivering conditioned air; and A sample chamber, comprising: a chamber housing having an upper face, a lower face opposite the upper face, and a wall extending vertically between the upper face and the lower face, wherein the wall defines an interior volume of the sample chamber, and the lower face is configured to face an imaging lens; and An air actuator unit is configured to direct conditioned air to a target location alongside the lower face to inhibit or at least reduce condensation accumulation on the imaging lens.

16. The system of claim 15 , wherein the air actuator unit comprises an air actuator, a conduit member, and an outlet port, wherein the air actuator is configured to introduce air from outside the sample chamber and direct the introduced air into a channel defined by the conduit member, wherein the channel is configured to direct the introduced air to the outlet port such that the introduced air exits the outlet port and travels outside and beside the second side of the chamber housing.

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

18. The system of claim 16 or claim 17, wherein the outlet port comprises fins elongated in respective directions diverging from one another in a downstream direction, thereby causing the incoming air to diffuse outwardly as it exits the outlet port.

19. The system of any one of claims 15 to 18, further comprising a cover, wherein the cover is configured for repeated attachment to and detachment from the upper face of the chamber housing, and the cover comprises at least one of a heating element or an insulating layer.

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

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