Thermal management of imaging system

Abstract

Managing thermal changes is important for obtaining quality data from optical systems. An optical system can include a light source (110), an objective lens (170) focused at a sample stage (185), and a transmission subassembly. The optical system can include a variety of lenses and mirrors configured to transmit light to the sample from there to an image forming device (195). Each of the transmission lenses, relay lenses, and relay objectives should be arranged in a predetermined alignment so that a multimode imaging module can function properly. A temperature measurement system (115) and an alignment control system (152) can be configured to control the alignment of these optical elements based on the measured temperature and calibrate for offsets, loss of focus, and other undesirable phenomenon that are otherwise introduced by temperature change.

Description

[0001] THERMAL MANAGEMENT OF IMAGING SYSTEM

[0002] CROSS-REFERENCE TO RELATED APPLICATIONS

[0003] The contents of the related applications filed on even date herewith entitled “Imager with Image Rotation” and “Automated Imaging in Content Screening” are hereby incorporated in their entirety by reference. Furthermore, the priority filings corresponding to these two applications, US 63 / 614,714 and US 63 / 614,743, are incorporated by reference herein in their entirety. This application claims the priority benefit of US 63 / 614,740, the contents of which are expressly incorporated herein by reference in their entirety.

[0004] BACKGROUND

[0005] Confocal fluorescence microscopy is a laser-based technique where radiation of one wavelength excites a fluorescent response in a sample that is detected at a second wavelength or range of wavelengths. Extensive libraries of fluorescent dyes have been developed to target different functional and structural elements of biological materials, for example cells, tissues, and organisms. Fluorescence microscopy enables researchers and clinicians to create, visualize and analyze micrographs of a sample where each color represents the distribution of specific target structures within the biological material. Various fluorescence microscopy techniques are described, for example in Renz, “Fluorescence Microscopy — A historical and Technical Perspective,” Cytometry Part A, Vol 83, pp. 767-779 (2013) and Sanderson et al., “Fluorescence Microscopy,” Cold Spring Harb Protoc. 2014(10): pdb.top071795. doi:10.1101 / pdb.top071795.

[0006] Complex microscopy and optical imaging systems may be affected by changes in temperature which may cause thermal drift in the optical paths of the various optical components of the imaging or microscopy systems. Thermal drift is challenging in light sheet systems, also referred to herein as selective plane illumination microscopy (SPIM), also referred to herein as oblique plane microscopy (OPM), because such systems require a number of lenses, mirrors, objectives, and tight alignments of multiple optical subassemblies in order to produce high contrast, high resolution images. SUMMARY

[0007] In one aspect, the technology relates to a thermal management device. The thermal management device includes a light source configured to produce an illumination light beam, a sample stage for receiving a sample, and an image acquisition device. An optical assembly is positioned along the optical path between the light source, the sample, and the image acquisition device. The optical assembly includes optical elements arranged in a predetermined alignment to direct the illumination light beam to the sample stage, and to direct a received light beam from the sample to the image acquisition device. A temperature measurement system is also included. An alignment control system can adjust an alignment of at least one of the optical elements in response to a change in temperature measured by the temperature measurement system.

[0008] The optical assembly can include an objective lens focused at the sample stage. The optical assembly can further include a transmission subassembly between the light source and the objective lens. The transmission subassembly can include at least one of a transmission lens and a mirror, and can direct the illumination light beam received from the light source to the sample and the received light beam from the sample to the image acquisition device. An imaging module can receive the light beam from the sample via the transmission assembly and transmit that received light beam to the image acquisition device via an ocular lens. The imaging module comprising a first alignment mechanism connected with a relay lens in a way that allows the first alignment mechanism to move the relay lens in response to a change in temperature. The imaging module also includes a second alignment mechanism coupled to a relay objective in a way that allows it to move the relay objective in response to the change in temperature.

[0009] The imaging module can be a multimode imaging module. There may also be a temperature control device functionally coupled to the temperature measurement system and configured to control a temperature within the optical assembly. The temperature control device can include a cooling element, a heating element, and a controller configured to control operation of the cooling element and the heating element to control the temperature within the optical assembly based on a difference between a benchmark temperature and a temperature measured by the temperature measurement system. The temperature measurement system can include one or more temperature measurement devices arranged to detect a temperature at one or more of the plurality of optical elements.

[0010] The alignment control system can include one or more alignment mechanisms coupled to each of the optical elements, a standard pattern arranged on the sample stage, the standard pattern comprising known features, and a processing device configured to determine a quality of an image of the standard pattern formed on the image acquisition device, and to adjust the alignment of at least one of the optical elements based on the determined quality of the image of the standard pattern. One or more alignment mechanisms are configured to translate, tilt, center, and focus at least one of the optical elements.

[0011] According to another aspect, a thermally managed imaging method of an optical assembly that comprises a plurality of optical elements and an image acquisition device, the plurality of optical elements being aligned in a predetermined alignment at a benchmark temperature. The method can include measuring a temperature of the optical assembly. When a difference between the benchmark temperature and the measured temperature of the optical assembly is equal to or above a first threshold, the method can include determining an adjustment to an alignment of at least one of the plurality of optical elements.

[0012] The method can include forming an image of a standard pattern formed on the sample stage at the image acquisition device in response to the difference being equal to or above the first threshold, and estimating a quality of the imaged standard pattern to determine the adjustment of the alignment of the at least one of the optical elements based on the estimated quality of the formed image. Based on these data, an adjustment can be both determined and performed either manually or automatically by the system.

[0013] The optical elements can include at least an objective lens, one or more transmission lenses, one or more relay lenses, one or more relay objectives, and an ocular lens. Performing the determined adjustment can include aligning at least one of the plurality of optical elements in an adjusted alignment that is different from the predetermined alignment. The adjustment that is performed can be one of translating, tilting, centering, and focusing at least one of the plurality of optical elements. For example, a temperature threshold can be in a range of 2° C to 10° C. Adjusting the alignment can include adjusting the alignment when the estimated quality of the imaged standard pattern is below a quality threshold so as to increase the quality of the formed image.

[0014] Estimating the quality of the imaged standard pattern can include accessing one optical characteristic from the group consisting of a benchmark resolution, a benchmark contrast, a signal level, an illumination position, and a benchmark focus of the standard pattern, and determining at least one optical characteristic from the group consisting of a resolution, a contrast, a signal level, an illumination position, and a focus of the imaged standard pattern, and comparing one of the determined optical characteristics to a corresponding one of the accessed optical characteristics.

[0015] The estimated quality of the imaged standard pattern can be below the quality threshold when the difference between the compared determined optical characteristic and the accessed optical characteristics is greater than a second threshold. The second threshold can be about 10%. Estimating the quality of the imaged standard pattern can include comparing features of the imaged standard pattern to a known set of features thereof. The estimated quality of the imaged standard pattern is below the quality threshold when a difference between sizes of the features of the imaged standard pattern and sizes of the known features is equal to or above an acceptable tolerance, which can be about 10%. Adjusting the alignment can include automatically adjusting the alignment of at least one of the plurality of optical elements based on the measured temperature of the optical subassembly.

[0016] The method can also include forming another image of the standard pattern at the image acquisition device after adjusting the alignment, and estimating a quality of the formed other image of the standard pattern. The method can include forming an image of a sample at the image acquisition device, by generating an illumination light beam at a light source, transmitting the illumination light beam to the objective lens via a transmission subassembly that includes the transmission lenses, transmitting, via at least one of the objective lens and the one or more transmission lenses, a received light beam to a multimode imaging module, the multimode imaging module comprising the one or more relay lenses and the one or more relay objectives, and transmitting the received light beam to the image acquisition device to form the image of the sample.

[0017] According to another aspect, an optical system includes the thermal management device as well as a communication device. One or more controllers can be operatively coupled to the imaging device and to the communication device. A memory can be coupled to the at least one controller, the memory storing instructions that, when executed by the controller, performs a set of operations. These operations can include measuring, via the temperature measurement system, a temperature of the optical assembly, and when a difference between a benchmark temperature and the measured temperature of the optical assembly is equal to or above a first threshold, determining an adjustment to an alignment of at least one of the plurality of optical elements.

[0018] The optical assembly can be configured to form an image of the standard pattern formed on the sample stage at the image acquisition device in response to the difference being equal to or above the first threshold, and estimate a quality of the imaged standard pattern to determine the adjustment of the alignment of the at least one of the plurality of optical elements based on the estimated quality of the formed image. The controller can be configured to determine a quality of the imaged standard pattern and adjust, via the alignment control system, an alignment of at least one of the plurality of optical elements based on the estimated quality of the formed image. The controller can be configured to perform the determined adjustment. The set of operations can include adjusting the alignment by aligning at least one of the plurality of optical elements in an adjusted alignment that is different from the predetermined alignment. The set of operations can also include adjusting the alignment by one of translating, tilting, centering, and focusing at least one of the optical elements.

[0019] The temperature measurement system can include one or more temperature measurement devices arranged to detect a temperature at one or more of the light source, the sample stage, and the plurality of optical elements. The set of operations can also include adjusting, via the alignment control system, an alignment of at least one of the plurality of optical elements based on the estimated quality of the formed image so as to increase the quality of the formed image when the estimated quality of the imaged standard pattern is below a quality threshold. The set of operations can include adjusting the alignment by automatically adjusting the alignment of at least one of the plurality of optical elements in an adjusted alignment that is different from the predetermined alignment.

[0020] BRIEF DESCRIPTION OF THE DRAWINGS

[0021] FIGS. 1A-E are schematic views of a multimode imaging system including SPIM, according to examples of this disclosure. FIG. 2A is a schematic view of a multimode imaging module, according to various examples of this disclosure.

[0022] FIG. 2B depicts an alignment subsystem including a plurality of alignment mechanisms, according to various examples of this disclosure.

[0023] FIG. 3 is a flow chart illustrating a method of thermally managing an optical imaging system, in accordance with various examples of the disclosure.

[0024] FIG. 4 depicts a block diagram of a computing device.

[0025] DETAILED DESCRIPTION

[0026] Complex microscopy and optical imaging systems may be affected by changes in temperature which may cause thermal drift in the optical paths of the various optical components of the imaging or microscopy systems. Thermal drift is challenging in optical systems, including for example light sheet systems, also referred to herein as selective plane illumination microscopy (SPIM) (also referred to herein as oblique plane microscopy (OPM)). SPIM / OPM systems require a significant number of (and in many cases moveable) lenses, mirrors, and objectives, along with tight alignments of multiple optical subassemblies in order to produce high contrast, high resolution images. SPIM is a technique for obtaining optically sectioned images of a sample. A SPIM optical system typically has two objective lenses, separated by an angle such as, e.g., an angle of 90°, relative to one another. Both objective lenses are used to view the same sample. For example, one lens may be used to illuminate only a thin “sheet” within the sample, and the second lens may be used to produce a diffraction-limited image of the same sheet.

[0027] Thermal drift may be expressed by slight changes in the position of any of the optical elements discussed above. The slight changes in the position of the optical elements may result in a deterioration of the quality of the image formed at the image acquisition device by, e.g., a deterioration of the focus or of the resolution thereof. Accordingly, thermal drift may be the cause of the deterioration of image quality at the image acquisition device. In order to remedy the image quality deterioration, one or more of the above optical elements may be repositioned / re-aligned, or their alignment may be adjusted. For SPIM, at a predetermined temperature, or within a predetermined temperature range, the optical components are arranged in a predetermined configuration (where the configuration comprises separation between elements, angle, tilt, orientation, etc.) to ensure alignment between the optical axes of the illuminated and detected planes. This minimizes blur generated by the rest of the sample, and by doing so achieves an optimized resolution, contrast etc. in a resulting image.

[0028] Due to thermal drift caused by increased temperature within a SPIM or other types of optical systems, a number of drawbacks constitute a hurdle to the accurate operation thereof. One of these drawbacks includes temperature drift, also referred to herein as thermal drift, which can cause a substantial misalignment of the SPIM or other type of optical system. Without temperature management, users may not obtain a sufficient performance from their SPIM or other type of optical system. This may be particularly true in settings where ambient temperature around the optical system fluctuates due to environmental or other conditions. In SPIM, the two objective lenses are typically required to remain properly aligned relative to each other, and this requirement may present a disadvantage at least because the alignment of the two objective lenses is typically difficult to achieve and to maintain, and even more so when thermal drift occurs. Another disadvantage is that it is typically difficult to arrange for the two objectives to be placed close enough to one another so that a high numerical aperture lens can be used to collect the light while still being able to produce a thin sheet of illumination, and as such a special sample holder may be needed to be used. Accordingly, thermal drift may present various challenges in obtaining a sufficient image quality, e.g., a sufficient image focus, contrast and / or resolution.

[0029] Examples of this disclosure are related to optical imaging systems. In particular, examples of this disclosure are related to managing thermal drift in optical imaging systems that include one or more methods and systems to characterize, monitor, and react to thermal changes, the thermal changes being internal to the optical system or external to the optical system, via an automated workflow in order to provide consistent imaging results to high content screening users. An apparatus according to various examples of the current disclosure may include a fully enclosed imaging system with cooling fans, heating elements, temperature sensors, control firmware or software, motorized optomechanical adjustment mechanisms, and software packages. These elements may be relied on to characterize, monitor, and react to thermal changes that may affect the optical alignment of optical elements within the system. A method according to examples of the disclosure may include characterizing or utilizing embedded calibration targets to measure the performance of the SPIM system at benchmark temperatures, and aligning the motorized adjustment positions (described below) to optimize performance at a range of expected temperatures. Such a method may also include monitoring the temperature of the SPIM system via temperature sensors to monitor any temperature changes between benchmark temperature(s) and current temperatures, whether internal temperatures or ambient temperatures. The method may also include reacting by, e.g., urging an automated workflow to analyze the measured temperature and to allow for user-selectable control methods. Such control methods include, but are not limited to, waiting for a steady state, heating or cooling the SPIM system, adjusting one or more optical mechanisms, and re-calibrating the optical alignment of one or more optical components of the SPIM system. In the case of SPIM, for example, the methods provide for the real-time dynamic reconfiguration of the various optical elements to maintain optimal alignment of the optical axes of the illuminated and detected planes.

[0030] FIGS. 1A-1E are schematic views of an imaging system 100, according to various examples of this disclosure. The imaging system 100 illustrated in FIGS 1A-1B are multimode imaging systems which include an optical arrangement that may be operated in one or more of a SPIM imaging mode 100 A, 100B (depicted in FIGS. 1A- B), a widefield imaging mode, a confocal imaging mode, or a brightfield mode. The imaging systems depicted in FIGS 1C-E are schematic views of single mode imaging systems, a widefield imaging mode 100C (depicted in FIG. 1C), a confocal imaging mode 100D (depicted in FIG. ID, with optional spinning disk 155), a brightfield mode 100E (depicted in FIG. IE). It will be appreciated that the multimode imaging systems of 100A, 100B may be configured to operate in the widefield imaging mode, a confocal imaging mode, or a brightfield mode, as illustrated in FIGS 1C-E. Similarly, it will be appreciated that an imaging system may be configured to operate in a SPIM-only imaging mode. No matter whether a single or multi-mode imaging system, each incorporates the teachings of this disclosure, managing thermal drift in optical imaging systems by providing one or more methods and systems to characterize, monitor, and react to thermal changes (the thermal changes being internal to the optical system or external to the optical system), via an automated workflow in order to provide consistent imaging results to high content screening users.

[0031] FIGS. 1A-1E are described concurrently and not every component described is depicted in every figure. FIGS. 1A-1E depict a light source 110 configured to generate an incident light “IL.” The light source 110 may be a laser, a light emitting diode (LED), an incandescent lamp, a halogen lamp, an arc lamp, and the like. FIGS. 1A-1E also depict a fiber selector 120 configured to direct the illumination light IL to a given or desired light path, filters 130 and 135, an ocular lens 190, and an image acquisition device 195 such as, e.g., at the camera. The fiber selector 120 may include a selectable first optical fiber 122 and a selectable second optical fiber 124, each selectable optical fiber 122 or 124 being configured to emit the illumination light IL therethrough. In examples, either or both of the first optical fiber 122 and the second optical fiber 124 may be or include a fiber bundle, the bundle including a plurality of fibers.

[0032] FIG. 1A-1E also describe a first lens 140 and a second lens 150, a mirror 160, an objective lens 170. The objective lens 170 may face a sample 180 arranged on a movable stage 185 that is movable in a planar or XY direction central to the objective lens 170. Details regarding the above components are described further below, as relevant to the various modes (brightfield, widefield, confocal, and SPIM). The controller 102 is configured to switch between the various modes illustrated in FIGS. 1A-1B and, e.g., select the optical fibers 122 and 124 in view of a desired imaging mode. Operation of at least one of the light source 110 or the fiber selector 120 is controlled via controller 102 to transmit the illumination light IL in a light sheet light path, the illumination light IL being referred to in this case as a first light source signal, or in a widefield or confocal light path, the illumination light IL being referred to in this case as a second light source signal. The controller 102 in FIGS. 1C-1E is configured to operate the light source, that is the light source 110 or the brightfield source 112.

[0033] The temperature monitoring and compensating technologies described herein, however, are equally applicable to imaging systems that include fewer than all four of these types of imaging modes, or imaging systems that include other types of imaging modes. Given the particular complexity of multimode imaging systems, however, the multimode imaging system 100A will be described herein for consistency and illustrative purposes.

[0034] FIG. 1 A depicts a light source 110 such as, e.g., a laser or LED, configured to generate an illumination light or incident light “IL,” a fiber selector 120 configured to direct the illumination light IL to a given or desired light path, filters 130 and 135, an ocular lens 190, and an image acquisition device 195 such as, e.g., a camera. The fiber selector 120 may include a selectable first optical fiber 122 and a selectable second optical fiber 124, each selectable optical fiber 122, 124 being configured to emit the illumination light IL therethrough towards a given optical path. FIG. 1A also depicts a first lens 140 and a second lens 150, a mirror 160, and an objective lens 170. The objective lens 170 may face a sample 180 arranged on a movable stage 185, and the sample 180 is movable via the movable stage 185 in a planar or XY direction that is parallel to the objective lens 170. The sample stage 185 may include a standard pattern 181, which may be a pattern formed in the sample stage 185 prior to operation, and any features of the pattern are known. Specifically, the sizes and shapes of the features included in the standard pattern 181 may be known under predetermined alignment conditions, for example at predetermined temperature(s). Accordingly, the standard pattern may be formed or disposed on the sample stage 185 and may include one or more features of known dimensions. The features may be or include, e.g., a grid pattern, a geometric shape with known sizes and shapes, other features of known sizes and shapes.

[0035] The controller 102 is configured to switch between various modes including the SPIM mode, the brightfield mode, widefield mode and the confocal mode and, e.g., select the optical fibers or optical fiber bundles 122 and 124 in view of a desired imaging mode. For example, the optical fiber 122 may be selected in the SPIM mode (FIGS 1 A-B), and the optical fiber 124 may be selected for the widefield mode (providing the optical paths illustrated in FIG 1C) or the confocal mode (providing the optical paths illustrated in FIG ID). In the brightfield imaging mode (as illustrated in FIG IE), sample illumination is transmitted from one side thereof and observed at an opposite side thereof. The illumination light IL is generated by a brightfield source 112 directly onto the sample 180 located at the movable stage 185. The illumination light IL emitted from the brightfield source 112 thus travels through the sample 180 and reaches the objective lens 170. In the imaging mode 100C the controller 102 is configured to, inter alia, control operation of the brightfield source 112. Operation of at least one of the light source 110, brightfield source 112 or the fiber selector 120 is controlled via the controller 102 to transmit the illumination light IL in, e.g., a SPIM light path, the illumination light IL being referred to in this case as the illumination light or illumination signal. The controller 102 may also include an alignment control subsystem, as described in more detail below, and especially in the context of FIG. 2B. The selectable first optical fiber 122 of the fiber selector 120 may be configured to emit the illumination signal therethrough towards the light sheet light path, and the selectable second optical fiber 124 may be configured to emit another light source signal therethrough towards, e.g., a widefield or confocal light path (FIGS. 1C, ID). The controller 102 may be coupled to, incorporated in or include a computing system such as, e.g., the computer system 400 discussed below with respect to FIG. 4.

[0036] In operation during, e.g., a SPIM mode, also referred to herein as light sheet mode, the light source 110 generates the illumination light IL that is transmitted to the sample 180 via the objective lens 170, the dichroic mirror 125 and the reflective mirror 160. When the sample 180 is illuminated by the IL, the sample 180 may emit a signal (an emission light), also referred to herein as returned light “RL.” An optical subassembly as defined herein can include fiber selector 120, dichroic mirror 125, mirror 160, and objective 170, though in alternative embodiments there may be more, fewer or different elements as part of the optical subassembly that accomplish the same effect of directing the illumination light IL to the sample at an angle.

[0037] For example, the returned light RL may be a fluorescent emission. Fluorescent emissions are the emissions of light from a sample at one wavelength (or a range of wavelengths) due to excitation at another wavelength. Fluorescent excitation and emission processes are a form of inelastic scattering of incident light and can be used to characterize a sample by providing information about the types of fluorescent emissions (number of photons emitted, and wavelength of emitted photons) based on a particular intensity and spectrum of incident light. “Autofluorescence” refers to fluorescence that occurs naturally upon exposure of a sample to an excitation source, while fluorescence more broadly can refer either to autofluorescence or to exogenous fluorescence via the application / integration of external fluorophores like fluorescent dyes, fluorescent proteins, and fluorescent nanoparticles or other fluorescence treatments.

[0038] The returned light RL may then be transmitted to the imaging mode selection module 175 and then to the image acquisition device 195 or camera via one or more lenses and mirrors such as, e.g., the first lens 140, the second lens 150, the filters 130 and 135, and the ocular lens, 190.

[0039] In the light sheet imaging system 100A, the imaging mode selection module 175 may be movable in the Z direction, (that is, orthogonal to the XY plane in which the stage 185 is movable as illustrated in FIG. 1A), and may be configured to be placed in the path of the returned light RL in order to convert the returned light RL from an off-centered light beam to the parallel light beam 178 that then projects onto ocular lens 190. For example, when the fiber selector 120 directs the illumination light IL to the dichroic mirror 125 in a light sheet mode, then the imaging mode selection module 175 is moved along the Z direction to be placed on the light path of the returned light RL. On the other hand, when the fiber selector directs the illumination light IL to the filter 130 and through a path corresponding to the light path of another imaging mode such as, e.g., a widefield or confocal mode (FIGS. 1C, ID) through the center of the filter 130, then the imaging mode selection module 175 may be moved off the light path of the returned light RL. In another example, the light path described above between the sample 180, the lens 170, the mirror 160, the dichroic mirror 125, the second lens 150, the first lens 140, the filters 130 and 135, the imaging mode selection module 175 and the ocular lens 190 may be referred to herein, or included within, as a transmission subassembly.

[0040] As noted above, temperature change or fluctuations in or around the imaging system 100A may cause thermal drift of components therein, leading to misalignments of the components and resulting undesirable imaging results. As such, the imaging systems described herein utilize an alignment control subsystem that includes a number of components to move (around or along various axes by translation, rotation, tilting, or combinations of those or other movements) the various elements along the light paths within the system 100A.

[0041] For example, the first lens 140 and / or the second lens 150 may be aligned along their optical axis via a lens / filter alignment mechanism 152 so as to independently adjust the alignment of the first lens 140, of the second lens 150, either using an internal processor or via instructions from the controller 102. The alignment mechanism 152 may include various linkages, gears, lead screws, gimbals, drives, motors, couplings, encoders, limit switches, or other components, as well as one or more controllers to control the various motors to enable the various types of movements of the various components that are contemplated herein. Further, while only a single alignment mechanism 152 is depicted for illustrative purposes, a plurality of such devices may be utilized. In examples, a discrete alignment mechanism may be utilized for each moveable lens or filter. In other examples, the same types of components, e.g., both lenses or both filters, may be adjusted by a discrete lens or filter alignment mechanism, respectively. Further, multiple alignment mechanisms may be used for complex movements of one or more components (e.g., a first alignment mechanism to move linearly a lens, and a second alignment mechanism to move rotatably the same lens). Other configurations of alignment mechanisms are contemplated. The various movements may include, for example, adjusting a distance between the second lens 150 and the first lens 140 along an optical axis, rotating one of the second lens 150 and the first lens 140, and / or tilting one of the second lens 150 and the first lens 140. The adjustment of any one or more of the distance between lenses 140 and 150, the tilt of one or both of the lenses 140 and 150, or the rotation or both of one of the lenses 140 and 150 may improve the quality, e.g., the focus, contrast and / or resolution of the image formed on the image acquisition device 195.

[0042] Other alignment mechanisms are included in the alignment control subsystem. For example, the objective lens 170 and the stage 185 may be independently movable and aligned via alignment mechanisms 172 and 182, respectively, the alignment mechanisms 172 and 182 being configured to independently adjust the alignment of the objective lens 170 and the stage 185 by independently translating, tilting and / or rotating the objective lens 170 and / or the stage 185. The alignment mechanisms 172 and 182 may independently include various linkages, gears, lead screws, gimbals, drives, motors, couplings, encoders, limit switches, or other components, as well as one or more controllers to control the various motors to enable the various types of movements of the various components that are contemplated herein. Further, while only single alignment mechanisms 172 and 182 are respectively depicted for illustrative purposes, a plurality of such devices may independently be utilized.

[0043] FIG IB illustrates multimode imaging system similar to that illustrated in Figure 1 A, but including another alignment mechanism, beam shaping optics or a beam steering device 127, for example, an adjustable mirror which is utilized to steer the direction of the illumination light IL. The beam steering device 127 may also be or include a spatial light modulator instead of an adjustable mirror. A “spatial light modulator” is a device that provides position addressable control over the amplitude and / or optical phase of a light beam that is reflected off of it or transmitted through it. A spatial light modulator can comprise a 2D array of electronically addressable variable retarders, including liquid crystal variable retarders. Spatial light modulators can also include reflective devices such as liquid crystal on silicon (LCOS), and MEMS based devices like micro-mirror array devices.

[0044] The beam steering device 127 may also be referred to herein as beam steering mirror 127, or pupil position adjustment mirror 127, and is configured to reflect the illumination light IL received form the fiber selector 120 onto various portions of the dichroic mirror 125. The imaging system 100B may also include an adjustment mechanism 129, which may include at least one motorized actuator to enable a translational movement of the beam steering device 127. In another example, the adjustment mechanism 129 may be configured to rotate the beam steering device 127 by an angle around an axis thereof. In yet another example, both rotational and translational movements may be performed by the adjustment mechanism 129. The adjustment mechanism 129 may be configured to translate the beam steering device 127 along a propagation direction “X” of the illumination light IL, or a direction “X” that includes a component that is parallel to the direction of the illumination light IL (see FIG. IB). Although the adjustment mechanism 129 is discussed above and illustrated in the drawings as a single mechanism, the adjustment mechanism 129 may be or include an individual rotation mechanism or an individual translation mechanism. This adjustment mechanism 129 provides an additional or alternative mechanism for implementing alignment of the various optical components to provide to facilitate the desired imaging result.

[0045] In a further example, the repositioning / alignment mechanism may comprise an optical redirection module, a combination of one or more mirrors, or comprise a prism.

[0046] When the temperature of the imaging system 100A changes, there may be resultant thermal drift in the position, angle of rotation, and / or tilt of any one or more of the lenses 140 and 150, the dichroic mirror 125, the reflective mirror 160, the objective lens 170, among other components of the imaging system 100A. Accordingly, the various alignment mechanisms such as alignment mechanisms 152, 172 and 182 described above may be controlled to adjust the position or alignment of any one or more of the above optical elements. The various alignment mechanisms such as alignment mechanisms 152, 172 and 182 may be operatively coupled to corresponding optical handling components that are disposed along the optical path of the illumination light IL or the optical path of the returned light RL, or both. Adjusting the alignment of any one or more of the above optical elements may correct, mitigate, reduce or remove any deterioration of the quality of the image formed on the image acquisition device 195 by, e.g., refocusing one or more of the optical elements in response to thermal drift.

[0047] Accordingly, one or more of the sample 180, the sample stage 185, and the optical elements including the objective lens 170, the mirror 160, the dichroic mirror 125, the second lens 150, the first lens 140, the filters 130 and 135, the imaging mode selection module 175 and the ocular lens 190 may be aligned in a predetermined alignment configured to image the sample 180 located on the sample stage 185 at, e.g., a given benchmark temperature. For example, the standard pattern 181 that is formed on the sample stage 185 may be used to test the predetermined alignment of optical elements. The predetermined alignment of the above optical elements may be formed based on imaging the standard pattern 181 with a sufficient level of contrast, focus, and / or resolution. Due to changes in temperature of the multimode imaging system 100A and / or in an environment thereof, the quality of the images formed on the image acquisition device 195 may be deteriorated due to thermal drift of the various optical elements discussed above.

[0048] One or more of the sample 180, the sample stage 185, the objective lens 170, the mirror 160, the dichroic mirror 125, the second lens 150, the first lens 140, and the ocular lens 190 may be movably controllable via the plurality of alignment mechanisms 152, 172 and 182 which may be, e.g., motorized devices or motors, and which may be configured to individually control a rotation, a translation, and a tilt of any one or more of the above optical elements (referred to herein as “optical handling elements”). The multimode imaging system 100A may also include one or more temperature sensors, strain gauges or other position sensors or encoders 115 located at one or more locations of the multimode imaging system 100A, and configured to measure the temperature of the multimode imaging system 100A, and of the environment in which various optical elements are located.

[0049] The multimode imaging system 100A may also include one or more temperature control devices 118. For example, the one or more temperature control devices 118 may include cooling fans, heating elements, refrigerating systems such as, e.g., cold liquids or gases flowing in insulated vessels throughout various portions of the multimode imaging system 100A. The temperature control devices 118 may be coupled to the temperature sensors 115 and to a controller such as controller 102. FIG. 2A shows the returned light RL received at a mirror 281 that redirects the returned light RL with respect to the initial light path of the returned light RL onto a first lens 282, also referred to herein as first relay lens 282. The returned light RL is received in a radially off-centered portion of the relay lens 282 and onto another relay lens 283, also in a radially off-centered portion thereof. The lens 283 then directs the returned light RL to a radially off-centered portion of a mirror 284 which redirects the returned light RL at an angle with respect to the incident returned light RL and towards a first relay objective 286. Other optical arrangements are possible, but importantly the returned light RL is transmitted such that it is received off-center at the first relay objective 286. The mapping of the pupils of first and second relay objectives 286 and 287 results in spatially-resolved data that can be used as described herein. Movement of the upper stage 204 affects the spatial position of the incoming returned light RL beam and these lenses map that incoming position to a 1 : 1 spatially-resolved position on the objectives 286, 287, either centered or offset depending upon the incoming returned light RL.

[0050] The first relay objective 286 transmits that redirected returned light RL to a second relay objective 287, the second relay objective 287 converts the radially off- centered returned light RL to a parallel light beam onto a mirror 289. The mirror 289 then redirects the received parallel light beam of the returned light RL out of the movable focusing module 200 as a parallel light beam 288. As a result, the returned light RL, initially entering the movable focusing module 200 as a radially off-centered light signal or light source signal, is converted to a centered parallel light beam 288 that is usable to an image acquisition device such as, e.g., the camera 195 illustrated in FIGS. 1 A-1E. The relay objectives 286 and 287 may be aligned and fixed in space on, e.g., a platform, to preserve the alignment therebetween.

[0051] The movable focusing module 200 may include a first portion 204 and a second portion 208. The first portion 204, also referred to herein as movable portion 204, includes the mirrors 281 and 289. The second portion 208, also referred to herein as fixed portion, includes a plurality of relay lenses such as, e.g., relay lenses 282, 283, and a plurality of relay objectives such as, e.g., relay objectives 286 and 287, and the mirror 284. The first portion 204 is configured to receive the returned light RL along a first axis, e.g., the axis of the returned light RL. Moving mechanism 279 is mechanically coupled to first portion 204 to cause translation thereof relative to second portion 208. Movement mechanism 279 can be controlled by, e.g., the controller 102 illustrated in FIGS. 1A-1E. In other examples, both the portions 204 and 208 may be movable via, e.g., the moving mechanism 279. In alternative examples, the portion 204 is fixed and the portion 208 is movable via, e.g., the moving mechanism 279. Although only two relay lenses 282 and 283 are depicted and discussed herein, more than two relay lenses (or other optical handling components) may be part of the movable focusing module 200, or part of the second portion thereof 208. 204 and 208 are independent but optically aligned to each other. First portion 204 can select whether light is transmitted through second portion 208 or not, and returns the output back into the widefield path, creating a multimodal system for use in OPM.

[0052] Accordingly, in operation, when the SPIM or light sheet mode is activated, the first portion 204 is placed on the light path of the optical signal or returned light RL on the optical axis of the ocular lens 190, and directs the returned light RL from the first portion 204 into the second portion 208 before returning to the first portion 204 and out of mirror 289 as parallel light beam 288. In particular, the second portion 208 receives, from the first portion 204 along an axis of the Z direction, the returned light RL at the first relay lens 282. The second portion 208 transmits the returned light RL from the relay lens 282 to the second relay lens 283 and to a first relay objective 286, then from the first relay objective 286 to the second relay objective 287, and then transmits the returned light RL to the first portion 204 from the second relay objective 287. The first portion 204 then transmits the returned light RL received from the second relay objective 287 to an image acquisition device 195 such as, e.g., the camera illustrated in FIGS. 1A-1E. The second portion 208 may include a moving mechanism 280 that is configured to adjust, typically along the Z direction or axis, a position of at least one of the first relay lens 282 and the second relay lens 283 based upon an instruction from the controller 102. As shown in FIG. 2 A, a sufficiently large face of the mirror 281 can receive returned light RL at a variety of Z positions, and translation of first portion 204 in the Y axis can also reflect incoming returned light RL to different spatially-resolved positions within second portion 208. Whether in the Z direction or some other direction, a controller (e.g., 102) can be used to implement relative movement between first portion 204 and second portion 208. Although two relay lenses 282 and 283 are illustrated in FIG. 2A, other relay lenses may be present in the second portion 208 to relay the returned light RL from the first relay lens 282 to the first relay objective 286.

[0053] The multimode SPIM module 200 may include a first portion or mode selection module 204, and a second portion or OPM module 208. The first portion or mode selection module 204, referred to herein as a movable portion, includes the mirrors 281 and 289. The second portion or OPM module 208, also referred to herein as fixed portion, includes a plurality of relay lenses such as, e.g., relay lenses 282, 283, and a plurality of relay objectives such as, e.g., relay objectives 286 and 287, and the mirror 284. It should be understood that mode selection module 204 is not limited to use in SPIM, and could be used for SPIM or any number of optical modules that could be assembled on an instrument.

[0054] The mode selection module 204 is configured to receive the returned light returned light RL along a first axis, e.g., the axis of the returned light RL, and may be movable via a moving mechanism 279 controlled by, e.g., the controller 102 illustrated and described with respect to FIGS. 1A-1E. In other examples, both the modules or portions 204 and 208 may be movable via, e.g., the moving mechanism 279. In alternative examples, the mode selection module 204 is fixed and the OPM module 208 is movable via, e.g., the moving mechanism 279. Although only two relay lenses 282 and 283 are depicted and discussed herein, more than two relay lenses may be part of the multimode SPIM module 200, or part of the OPM module thereof 208. One or more of the relay lenses 282 and 283, the relay objectives 286 and 288, and the mirrors 281, 284 and 289 may be aligned via an alignment mechanism 280, and may be aligned according to a predetermined alignment in order to obtain an image of the sample 180, or of the standard pattern 181 form on the sample stage 185 (see FIGS. 1A-1E) of sufficiently high focus score, which is to say focus, contrast, and / or resolution that exceed some predetermined threshold.

[0055] The relay lenses 282 and 283 as well as the relay objectives 286 and 287, and the mirrors 281, 284 and 289 may be independently movable and aligned via, e.g., alignment mechanisms 271 and 273. For example, the alignment mechanisms 271 and 273 may be configured to independently adjust the alignment of the relay lenses 282 and 283 as well as the relay objectives 286 and 287, and the mirrors 281, 284 and 289 by independently translating, tilting or rotating any one of relay lenses 282 and 283 as well as the relay objectives 286 and 287, and the mirrors 281, 284 and 289. Accordingly, when the temperature of the multimode SPIM module 200 changes to the point of creating thermal drift in the position, angle of rotation, or tilt of any of the relay lenses 282 and 283, the relay objectives 286 and 287, and / or the mirrors 281, 284 and 289, the various alignment mechanisms such as alignment mechanisms 271 and 273 may be independently controlled to independently adjust the alignment thereof. Adjusting the alignment of any of the above optical elements may correct, mitigate, reduce or remove any deterioration of the quality of the image formed on the image acquisition device 195 illustrated in FIGS. 1A-1E by, e.g., refocusing one or more of the optical elements.

[0056] In operation, as changes in the temperature of the module 200 or the environment thereof, or the temperature of the imaging device 100A or the environment thereof, occur over time, one or more of optical elements may experience thermal drift. The optical elements discussed above include at least the relay lenses 282 and 283, the relay objectives 286 and 287, the mirrors 281, 284 and 289, the sample stage 185, the lens 170, the mirror 160, the dichroic mirror 125, the second lens 150, the first lens 140, the filters 130 and 135, the imaging mode selection module 175, and the ocular lens 190. The module 200 may also include one or more temperature sensors 215 located therein or adjacent thereto, and configured to measure the temperature of the module 200, and of the environment in which various optical elements are located.

[0057] FIG. 2B depicts an alignment subsystem including a plurality of alignment mechanisms, according to various examples of this disclosure. In FIG. 2B, the alignment subsystem 205 includes a processor 210. For example, the processor 210 may be similar to the processor 400 discussed below or the controller 102, and may be configured to control operation of a one or more of a plurality of alignment mechanisms such as the alignment mechanisms, 129, 152, 172, 182, 271, 273 and 280 discussed throughout the current disclosure. Each of the alignment mechanisms may be or include various linkages, gears, lead screws, gimbals, drives, motors, couplings, encoders, limit switches, or other components, as well as one or more controllers to control the various motors to enable the various types of movements of the various components that are contemplated herein.

[0058] In FIG. 2B, the alignment mechanism 152 may be configured to align, e.g., one or more lenses such as the lenses 140 and 150 illustrated in FIGS. 1A-1E. The alignment mechanism 172 may be configured to align, e.g., one or more objectives such as the objective 170 illustrated in FIGS. 1A-1E. The alignment mechanism 182 may be configured to align, e.g., the sample 180 illustrated in FIGS. 1A-1E. The alignment mechanism 271 may be configured to align, e.g., one or more lenses such as the lenses 282 and 283 illustrated in FIG. 2A. The alignment mechanism 273 may be configured to align, e.g., one or more objectives such as the objectives 286 and 287 illustrated in FIG. 2A. The alignment mechanism 280 may be configured to align, e.g., one or more mirrors such as the mirrors 281 and 289 illustrated in FIG. 2A. Although various alignment mechanisms are described as being configured to align specific optical elements, the alignment mechanisms may be configured to align other optical elements, and other similar alignment mechanisms may be configured to align the above-described optical elements. The controller 210 may also be coupled to the temperature sensor(s) 115 and to the temperature control device(s) 118. In operation, the temperature measured or sensed by one or more of the temperature sensor(s) 115 may be used to adjust the alignment of any of the above optical elements. Alternatively, or in addition, the temperature control device(s) 118 may be used to control the temperature of the imaging system 100A based on the temperature measured or sensed from the temperature sensor(s) 115. In a further alternative or additionally, the temperature measured or sensed by a temperature sensor may be used to trigger an automatic reconfiguration of the optical elements, or suggest recalibration. In some configurations, the suggestion may be conveyed to a user via a user interface, and the user be prompted to either authorize recalibration or defer recalibration at that time. In other configurations the system may be rendered inoperable, or the user may be prompted to wait until a temperature sensed by the one or more temperature sensor(s) 115 senses that the temperatures is within an acceptable range of the benchmark temperature. Any input provided by the user may be routed to the controller 102 to control the system accordingly.

[0059] FIG. 3 depicts an imaging method 300 that includes operation 310 which includes accessing a benchmark temperature of an optical assembly or imaging device via, e.g., a communication device or an input device. In some instances, the benchmark temperature may be a specific temperature range, for example within + / - 1 degree Celsius or + / - 2 degrees Celsius of the benchmark temperature. The optical assembly or imaging device may include a plurality of optical elements (that is, any combination of one or more objective lenses, transmission lenses, dichroic mirrors or other beam splitters or combiners, mirrors, filters, ocular lenses, relay lenses, relay objectives). The benchmark temperature may be a calibration temperature of the optical elements discussed above in a predetermined alignment. Accordingly, the image of a standard pattern such as, e.g., the standard pattern 181 illustrated in FIGS. 1A-1E, that is formed by the optical elements aligned in the predetermined alignment at the benchmark is deemed to be of sufficient quality, e.g., of sufficient focus, contrast, resolution, signal level, illumination position, and / or uniformity. Thus, during operation 310, the benchmark temperature for the optical assembly is accessed. In addition to accessing the benchmark temperature, operation 310 also includes accessing or determining a benchmark image quality such as, e.g., a one or more of a benchmark contrast, a benchmark resolution, and / or a benchmark focus. During operation 320, which may occur some amount of time after the optical elements have been aligned according to the predetermined alignment, the temperature of the optical assembly, referred to as 100 or 200 in FIGS. 1 and 2, may be measured. For example, the temperature may be measured via one or more temperature measurement devices. With reference to FIGS. 1A-1E, 2A, and 2B, the temperature measurement devices may be or include temperature sensors 115 and 215. Temperature monitoring may be continuous, for example 1Hz, and the operation 320 can be initiated by deviations in temperature as set by the user or defaulted to for example + / - 2 degrees Celsius.

[0060] During operation 330, a comparison is made between the benchmark temperature accessed during operation 310 and the measured temperature measured during operation 320. If the difference between the benchmark temperature and the measured temperature of the optical assembly is below a first threshold, then the method repeats operation 320, where the temperature of the optical assembly continues to be monitored and measured. If the difference between the benchmark temperature and the measured temperature of the optical assembly is equal to or above the first threshold, then the method continues to operation 340, where the standard pattern may be imaged. The first threshold may be in a range of about 2 °C to about 10 °C. The standard pattern may be the standard pattern 181 discussed above with reference to FIGS. 1A-1E and may be formed or disposed on, e.g., the sample stage 185, and may include one or more features of known dimensions. The features may be or include, e.g., a grid pattern, a geometric shape with known sizes and shapes, other features of known sizes and shapes. As such, the resulting image formed at the image acquisition device may have a reliable and expected focus, resolution, and / or contrast.

[0061] Operation 350 includes estimating the quality of the imaged standard pattern. Estimating the quality of the imaged standard pattern during operation 350 may include accessing the benchmark quality, and performing a comparison between the quality of the imaged standard pattern just obtained and the benchmark quality of the standard pattern under calibrated conditions. Accessing the benchmark quality of the imaged standard pattern during operation 350 may include, for example, accessing one of the benchmark resolution, the benchmark contrast, the benchmark focus of the standard pattern, signal level, illumination position, or uniformity. Comparing the benchmark quality may also include determining one of a resolution, a contrast, and a focus of the image of the standard pattern formed during operation 340, and comparing one of the determined resolution to the accessed benchmark resolution, the determined contrast to the accessed benchmark contrast, and the determined focus to the accessed benchmark focus. The estimated quality of the imaged standard pattern during operation 350 may be determined to be below the quality threshold when a difference between the determined and the benchmark resolution, contrast, and / or focus of the standard pattern is greater than an acceptable tolerance. Alternatively, estimating the quality of the imaged standard pattern may include comparing features of the imaged standard pattern to the known features. The estimated quality of the imaged standard pattern may be determined to be below the quality threshold when the difference between sizes of the features of the imaged standard pattern and sizes of the known features of the standard pattern is equal to or above the acceptable tolerance. The acceptable tolerance may be less than or equal to 10%.

[0062] Thus, as described above, estimating the quality of the imaged standard pattern can include accessing a chosen optical characteristic such as a benchmark resolution, a benchmark contrast, a signal level, an illumination position, and a benchmark focus of the standard pattern. The systems and methods can determine at least one optical characteristic such as a resolution, a contrast, a signal level, an illumination position, and a focus of the imaged standard pattern. The determined optical characteristic can be compared to the accessed optical characteristic.

[0063] Operation 360 includes comparing the quality of the imaged standard pattern to the benchmark quality. If during operation 360, the quality of the imaged standard pattern estimated during operation 350 is at or above a quality threshold, also referred to herein as second threshold, then the method continues to operation 370. During operation 370, the method may end, or the image of a sample may be formed without performing any further adjustment, re-positioning or re-alignment of any of the optical elements forming the optical assembly or imaging device. If during operation 360, the quality of the imaged standard pattern estimated during operation 350 is below the second threshold, then the method continues to operation 380.

[0064] During operation 380, the alignment of at least one of the optical elements discussed above is adjusted so as to increase the quality of the imaged standard pattern in order to correct for thermal drift of the alignment of the optical elements. In another example, adjusting the alignment may be performed automatically on at least one of the plurality of optical elements of the optical subassembly, and includes translating, tilting, rotating or focusing any one or more of the optical elements via a plurality of motorized devices under control of a processor such as, e.g., the processor 400 further discussed below. In an example of operation, when the difference between the ambient temperature and the benchmark temperature is greater than the first threshold, the standard pattern may be automatically imaged. When the quality of the imaged standard pattern is below the quality threshold or second threshold, one or more of the optical elements may automatically adjusted or aligned via the motorized devices under control of the processor. The motorized device may include, e.g., the alignment mechanisms 152, 172 and 182 discussed above with respect to FIGS. 1A-1E.

[0065] Accordingly, the alignment of one or more of the optical elements is adjusted during operation 380 so as to increase the quality of the formed image when that quality has been deteriorated by thermal drift and, as a result, the estimated quality of the imaged standard pattern is below the quality threshold, as determined during operation 360. The adjustment of the plurality of optical elements may include aligning at least one of the plurality of optical elements in an adjusted alignment, or realignment, that is different from the predetermined alignment. As indicated above, the automatic adjustment of the plurality of optical elements may be in the form of any one of an automatic translating, tilting, centering, and focusing one or more of the plurality of optical elements under control of a processor.

[0066] Automatic adjustment may be based on the use of look-up tables which identifying the various settings of the optical elements based on the determined temperature. In an alternative, automatic adjustment may comprise an iterative process of repositioning one or more, or a subset of the plurality of optical elements. For example, this may include scanning the sample in the Z direction by moving the primary objective along the optical axis while adjusting to a pre-determined light sheet remote focus position, pupil position, and / or steering for optimal performance. The scanning may be performed in a stepwise fashion, or a continuous scan, and similarly the light sheet adjustments may be controlled to move at each step or continuously synchronized to interpolated positions from pre-determined settings. Alternatively, the sample may be scanned in a layer-wise fashion in a direction perpendicular to the Z direction in a stepwise fashion or as a continuous scan by utilizing the sample stage, and at each layer, pre-determined adjustments may be performed to remote focus, pupil position and / or steering to achieve increased or optimal performance. In one example, software can collect images with variable adjustment settings in focus, pupil position and steering, and rank the images based on a determined focus score based on contrast metrics for example. The system settings associated with the highest-ranking images can be saved as adjustment positions, to be used for the final imaging.

[0067] In another alternative, automatic adjustment may be based on prior performance, or historical data. The prior performance or historical data being retrieved from the one or more volatile memory(ies) 406 or other dynamic memory component(s) of computing device 400, (as described below).

[0068] Subsequently to operation 380, once the re-alignment has been performed, the operation may revert to operation 340 to form another image of the standard pattern at the image acquisition device in order to determine whether the re-alignment has sufficiently improved the quality of the formed image and has thus correct any deleterious effect caused by thermal drift. Forming the image of the standard pattern after re-alignment during operation 340 may also be followed by an estimation of the quality of the imaged standard pattern during operation 350, as well as operations 360 and 380 if needed. The loop of operations 340, 350, 360 and 380, referred to feedback loops, may be repeated as needed, and for example may be automatically repeated as needed, until the quality of the imaged standard pattern is estimated during operation 360 to be sufficient, in which case the method continues to operation 370. Once the optical elements are aligned so as to produce an image of sufficient quality, the method may include forming an image of a sample at the image acquisition device during operation 370. Forming the image during operation 370 may include generating a light source signal at a light source, transmitting the generated light source signal to the objective lens in proximity to the sample via one or more transmission lenses, transmitting, via at least one of the objective lens and the transmission lenses, a sample signal generated by the sample to a multimode imaging module, and transmitting the sample signal to the image acquisition device to form the image of the sample. For example, the multimode imaging module includes one or more relay lenses and one or more relay objectives.

[0069] In another mode of operation repositioning or realigning of the optical elements in the optical path of the illumination light IL can performed independently from repositioning or realigning of the optical elements in the optical path of the returned light RL. In one embodiment, for example, a multimode optical system is operated in the widefield mode and the optical mechanism(s) utilized to reposition the optical elements in the reflective light optical path, prior to the multimode optical system being operating using a light sheet or SPIM mode, and optical mechanism(s) utilized to reposition the optical elements in the illumination path. Performing these two operations independently prior to rechecking the calibration target.

[0070] FIG. 4 depicts a block diagram of a computing device 400 configured to control the multimode imaging device 100A discussed above with respect to FIG. 1 A. hi the illustrated example, the computing device 400 may include a bus 402 or other communication mechanism of similar function for communicating information, and at least one processing element 404 (collectively referred to as processing element 404) coupled with bus 402 for processing information. As will be appreciated by those skilled in the art, the processing element 404 may include a plurality of processing elements or cores, which may be packaged as a single processor or in a distributed arrangement. Furthermore, a plurality of virtual processing elements 404 may be included in the computing device 400 to provide the control or management operations for, e.g., the multimode imaging device 100A illustrated above.

[0071] The computing device 400 may also include one or more volatile memory(ies) 406, which can for example include random access memory(ies) (RAM) or other dynamic memory component(s), coupled to one or more busses 402 for use by the at least one processing element 404. Computing device 400 may further include static, non-volatile memory(ies) 408, such as read only memory (ROM) or other static memory components, coupled to busses 402 for storing information and instructions for use by the at least one processing element 404. A storage component 410, such as a storage disk or storage memory, may be provided for storing information and instructions for use by the at least one processing element 404. As will be appreciated, the computing device 400 may include a distributed storage component 412, such as a networked disk or other storage resource available to the computing device 400.

[0072] The computing device 400 may be coupled to one or more displays 414 for displaying information to a user. Optional user input device(s) 416, such as a keyboard and / or touchscreen, may be coupled to Bus 402 for communicating information and command selections to the at least one processing element 404. An optional cursor control or graphical input device 418, such as a mouse, a trackball or cursor direction keys for communicating graphical user interface information and command selections to the at least one processing element. The computing device 400 may further include an input / output (I / O) component, such as a serial connection, digital connection, network connection, or other input / output component for allowing intercommunication with other computing components and the various components of, e.g., the multimode imaging device 100A discussed above.

[0073] In various examples, computing device 400 can be connected to one or more other computer systems via a network to form a networked system. Such networks can for example include one or more private networks or public networks, such as the Internet. In the networked system, one or more computer systems can store and serve the data to other computer systems. The one or more computer systems that store and serve the data can be referred to as servers or the cloud in a cloud computing scenario. The one or more computer systems can include one or more web servers, for example. The other computer systems that send and receive data to and from the servers or the cloud can be referred to as client or cloud devices, for example. Various operations of, e.g., multimode imaging device 100A may be supported by operation of the distributed computing systems.

[0074] The computing device 400 may be operative to control operation of the components of the multimode imaging device 100A through a communication device such as, e.g., communication device 420, and to handle data generated by components of the multimode imaging device 100A through the processing element 404. In some examples, feedback in the form of an image is provided by the computing device 400 in response to the processing element 404 executing instructions contained in memory 406 or 408 and performing operations on data received from the multimode imaging device 100A such as, e.g., receiving and transmitting optical signals and adjusting the alignment of the various optical elements via alignment mechanisms 152, 172 and 182. Execution of instructions contained in memory 406 and / or 408 by the at least one processing element 404 can render, e.g., the multimode imaging device 100A to perform methods described herein.

[0075] The term “computer-readable medium” as used herein refers to any media that participates in providing instructions to the processing element 404 for execution. Such a medium may take many forms, including but not limited to, non-volatile media, volatile media, and transmission media. Non-volatile media includes, for example, optical or magnetic disks, such as disk storage 410. Volatile media includes dynamic memory, such as memory 406. Transmission media includes coaxial cables, copper wire, and fiber optics, including the wires that include bus 402.

[0076] Common forms of computer-readable media or computer program products include, for example, a floppy disk, a flexible disk, hard disk, magnetic tape, or any other magnetic medium, a CD-ROM, digital video disc (DVD), a Blu-ray Disc, any other optical medium, a thumb drive, a memory card, a RAM, PROM, and EPROM, a FLASH-EPROM, any other memory chip or cartridge, or any other tangible medium from which a computer can read.

[0077] Various forms of computer readable media may be involved in carrying one or more sequences of one or more instructions to the processing element 404 for execution. For example, the instructions may initially be carried on the magnetic disk of a remote computer. The remote computer can load the instructions into its dynamic memory and send the instructions over a telephone line using a modem. A modem local to computing device 400 can receive the data on the telephone line and use an infra-red transmitter to convert the data to an infra-red signal. An infra-red detector coupled to bus 402 can receive the data carried in the infra-red signal and place the data on bus 402. Bus 402 carries the data to memory 406, from which the processing element 404 retrieves and executes the instructions. The instructions received by memory 406 and / or memory 408 may optionally be stored on storage device 410 either before or after execution by the processing element 404. In accordance with various examples, instructions operative to be executed by a processing element to perform a method are stored on a computer-readable medium. The computer-readable medium can be a device that stores digital information. For example, a computer-readable medium includes a compact disc read-only memory (CD-ROM) as is known in the art for storing software. The computer-readable medium is accessed by a processor suitable for executing instructions configured to be executed.

[0078] This disclosure described some examples of the present technology with reference to the accompanying drawings, in which only some of the possible examples were shown. Other aspects can, however, be embodied in many different forms and should not be construed as limited to the examples set forth herein. Rather, these examples were provided so that this disclosure was thorough and complete and fully conveyed the scope of the possible examples to those skilled in the art.

[0079] Although specific examples were described herein, the scope of the technology is not limited to those specific examples. One skilled in the art will recognize other examples or improvements that are within the scope of the present technology. Therefore, the specific structure, acts, or media are disclosed only as illustrative examples. Examples according to the technology may also combine elements or components of those that are disclosed in general but not expressly exemplified in combination, unless otherwise stated herein. The scope of the technology is defined by the following claims and any equivalents therein.

Claims

CLAIMSWhat is claimed is:

1. A thermal management device comprising: a light source configured to produce an illumination light beam; a sample stage for receiving a sample; an image acquisition device; an optical assembly between the light source, the sample and the image acquisition device, the optical assembly comprising a plurality of optical elements arranged in a predetermined alignment, the optical elements configured to direct the illumination light beam to the sample stage and a received light beam from the sample to the image acquisition device; a temperature measurement system; and an alignment control system configured to adjust an alignment of at least one of the plurality of optical elements in response to a change in temperature measured by the temperature measurement system.

2. The thermal management device of claim 1, wherein the optical assembly comprises: an objective lens focused at the sample stage; an optical subassembly between the light source and the objective lens, the optical subassembly comprising at least one of a transmission lens and a mirror, the optical subassembly being configured to transmit the illumination light beam received from the light source to the sample and the received light beam from the sample to the image acquisition device; an imaging module configured to receive the received light beam emitted from the sample via the transmission assembly and to transmit the received light beam to the image acquisition device via an ocular lens, the imaging module comprising a first alignment mechanism coupled to a relay lens and a second alignment mechanism coupled to a relay objective, wherein the first and second alignment mechanisms are configured to move the relay lens and relay objective, respectively, in response to the change in temperature.

3. The thermal management device of claim 1 or claim 2, wherein the imaging module is a multimode imaging module.

4. The thermal management device of any of claims 1-3, further comprising a temperature control device functionally coupled to the temperature measurement system and configured to control a temperature within the optical assembly.

5. The thermal management device of claim 4, wherein the temperature control device comprises a cooling element, a heating element, and a controller configured to control operation of the cooling element and the heating element to control the temperature within the optical assembly based on a difference between a benchmark temperature and a temperature measured by the temperature measurement system.

6. The thermal management device of any preceding claim, wherein the temperature measurement system comprises one or more temperature measurement devices arranged to detect a temperature at one or more of the plurality of optical elements.

7. The thermal management device of any preceding claim, wherein the alignment control system comprises: one or more alignment mechanisms coupled to each of the plurality of optical elements; a standard pattern arranged on the sample stage, the standard pattern comprising known features; and a processing device configured to determine a quality of an image of the standard pattern formed on the image acquisition device, and to adjust the alignment of at least one of the optical elements based on the determined quality of the image of the standard pattern.

8. The thermal management device of claim 7, wherein the one or more alignment mechanisms are configured to translate, tilt, center, and focus at least one of the optical elements.

9. An optical system comprising:the imaging device of any of claims 1-8; a communication device; at least one controller operatively coupled to the image acquisition device and to the communication device; and a memory coupled to the at least one controller, the memory storing instructions that, when executed by the controller, performs a set of operations comprising: measuring, via the temperature measurement system, a temperature of the optical assembly; when a difference between a benchmark temperature and the measured temperature of the optical assembly is equal to or above a first threshold, determining an adjustment to an alignment of at least one of the plurality of optical elements.

10. The optical system of claim 9, wherein the optical assembly is configured to form an image of a standard pattern formed on the sample stage at the image acquisition device in response to the difference being equal to or above the first threshold, such that a user may estimate a quality of the imaged standard pattern to determine the adjustment of the alignment of the at least one of the plurality of optical elements based on the estimated quality of the formed image.

11. The optical system of claim 10, wherein the controller is configured to determine a quality of the imaged standard pattern and adjust, via the alignment control system, an alignment of at least one of the plurality of optical elements based on the estimated quality of the formed image.

12. The optical system of any of claims 9-11, wherein the set of operations comprises adjusting the alignment by aligning at least one of the plurality of optical elements in an adjusted alignment that is different from the predetermined alignment.

13. The optical system of any of claims 9-12, wherein the set of operations comprises adjusting the alignment by one of translating, tilting, centering, and focusing at least one of the plurality of optical elements.

14. The optical system of any of claims 9-13, wherein the temperature measurement system comprises one or more temperature measurement devices arranged to detect a temperature at one or more of the light source, the sample stage, and the plurality of optical elements.

15. The optical system of any of claims 9-14, wherein the set of operations comprises, when the estimated quality of an imaged standard pattern is below a quality threshold, adjusting, via the alignment control system, an alignment of at least one of the plurality of optical elements based on the estimated quality of the formed image so as to increase the quality of the formed image.

16. The optical system of any of claims 9-15, wherein the set of operations comprises adjusting the alignment by automatically adjusting the alignment of at least one of the plurality of optical elements in an adjusted alignment that is different from the predetermined alignment.

17. A thermally managed imaging method of an optical assembly that comprises a plurality of optical elements and an image acquisition device, the plurality of optical elements being aligned in a predetermined alignment at a benchmark temperature, the method comprising: measuring a temperature of the optical assembly; and when a difference between the benchmark temperature and the measured temperature of the optical assembly is equal to or above a first threshold, determining an adjustment to an alignment of at least one of the plurality of optical elements.

18. The method of claim 17, further comprising forming an image of a standard pattern formed on a sample stage at the image acquisition device in response to the difference being equal to or above the first threshold, and estimating a quality of the imaged standard pattern to determine the adjustment of the alignment of the at least one of the plurality of optical elements based on the estimated quality of the formed image.

19. The method of claim 18, further comprising performing the determined adjustment.

20. The method of any of claims 17-19, wherein the plurality of optical elements comprise at least an objective lens, one or more transmission lenses, one or more relay lenses, one or more relay objectives, and an ocular lens.

21. The method of claim 18, wherein performing the determined adjustment comprises aligning at least one of the plurality of optical elements in an adjusted alignment that is different from the predetermined alignment.

22. The method of claim 19 or claim 21, wherein the adjusting comprises one of translating, tilting, centering, and focusing at least one of the plurality of optical elements.

23. The method of any of claims 18-22, wherein the first threshold is in a range of 2° C to 10° C.

24. The method of claims 19, 21, or 22, wherein the adjusting the alignment comprises adjusting the alignment when the estimated quality of the imaged standard pattern is below a quality threshold so as to increase the quality of the formed image.

25. The method of claim 24, wherein estimating the quality of the imaged standard pattern comprises: accessing one optical characteristic from the group consisting of a benchmark resolution, a benchmark contrast, a signal level, an illumination position, and a benchmark focus of the standard pattern; determining at least one optical characteristic from the group consisting of a resolution, a contrast, a signal level, an illumination position, and a focus of the imaged standard pattern; and comparing one of the determined optical characteristics to a corresponding one of the accessed optical characteristics.

26. The method of claim 25, wherein the estimated quality of the imaged standard pattern is below the quality threshold when the difference between the compareddetermined optical characteristic and the accessed optical characteristics is greater than a second threshold.

27. The method of claim 26, wherein the second threshold is less than or equal to 10%.

28. The method of any of claims 18-27, wherein estimating the quality of the imaged standard pattern comprises comparing features of the imaged standard pattern to a known set of features thereof.

29. The method of claim 28, wherein the estimated quality of the imaged standard pattern is below the quality threshold when a difference between sizes of the features of the imaged standard pattern and sizes of the known features is equal to or above an acceptable tolerance.

30. The method of claim 29, wherein the acceptable tolerance is less than or equal to 10%.

31. The method of any of claims 17-30, wherein adjusting the alignment comprises automatically adjusting the alignment of at least one of the plurality of optical elements based on the measured temperature of the optical subassembly.

32. The method of any of claims 19, 24, 28, 24-31, further comprising: forming another image of the standard pattern at the image acquisition device after adjusting the alignment; and estimating a quality of the formed other image of the standard pattern.

33. The method of any of claims 17-32, further comprising using the optical elements for forming an image of a sample at the image acquisition device, wherein forming the image of the sample using the optical elements comprises: generating an illumination light beam at a light source; transmitting the illumination light beam to the objective lens via an optical subassembly that comprises the one or more transmission lenses;transmitting, via at least one of the objective lens and the one or more transmission lenses, a received light beam to a multimode imaging module, the multimode imaging module comprising one or more relay lenses and one or more relay objectives; and transmitting the received light beam to the image acquisition device to form the image of the sample.

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