Sealed fluorescence microscope with constant beam path

Abstract

A fluorescence microscope includes a housing having an upper wall with an opening. A glass deck overlies the opening and is configured to seal the interior of the housing from an external environment. The glass deck includes an array of windows, and each window is configured to receive a respective specimen slide. A camera, a light source, and an optical assembly are provided within the housing. The light source is configured to generate an illumination light beam, and the optical assembly is movable to provide an optical path from the light source to the illumination light beam on a specimen positioned at each window in the array and to provide an optical path from each specimen to a fluorescence emission light beam to the camera. The optical assembly is configured such that the optical path lengths from each specimen to the camera for the fluorescence emission light beam are the same.

Description

[0001] Related applications

[0002] This application claims the benefit and priority of U.S. Patent Application No. 18 / 500,859, filed November 2, 2023, the disclosure of which is incorporated herein by reference in its entirety. Technical Field

[0003] The present invention generally relates to microscopes, and more specifically to fluorescence microscopes. Background Technology

[0004] Microscopes in incubator environments can become contaminated due to the conditions within the incubator. For example, microscopes in incubator environments may become moldy, accumulate pathogens or bacteria, and / or fungi may grow on them. Furthermore, conventional incubator microscopes are typically automated inverted microscopes, which can be bulky due to the size of their specimen scanning stages. Summary of the Invention

[0005] According to some embodiments of the present invention, a fluorescence microscope includes a housing having an upper wall with an opening. A glass stage covers the opening and is configured to seal and isolate the interior of the housing from an external environment, such as an incubator environment. The glass stage includes an array of windows, each window being configured to receive a corresponding specimen plate. A camera, a light source, and optical components are provided within the housing and protected by the housing from external environmental influences. The light source is configured to generate an illumination beam, and the optical components are movable to provide an optical path from the light source to the illumination beam positioned at each window in the array, and to provide an optical path for a fluorescence emission beam from each specimen to the camera. The optical components are configured such that the length of the optical path for the fluorescence emission beam from each specimen to the camera is substantially the same.

[0006] In some embodiments, the optical assembly includes a base fixed to the lower wall of the housing, a first arm movably fixed to the base, and a second arm movably fixed to the first arm. However, it should be understood that embodiments of the inventive concept are not limited to optical assemblies with only two arms. An optical assembly can have any number of arms, more than two. For example, the upper arm or the second arm can be split into two arms with two additional mirrors without altering the functionality of the optical assembly, merely increasing the height of the instrument. The first arm is configured to rotate about a first axis, and the second arm is configured to rotate about a second axis, which is spaced apart from and substantially parallel to the first axis. Each of the first and second arms includes optical components that generate an optical path for a fluorescence emission beam from each specimen to the camera. The second arm includes an objective lens, a beam splitter, and a first mirror. The beam splitter is configured to direct an illumination beam from a light source through the objective lens to the corresponding specimen, and to direct a fluorescence emission beam received by the objective lens from the corresponding specimen through the second arm to the first mirror. The first mirror is configured to direct the fluorescence emission beam into the first arm. The first arm includes a second mirror and a third mirror. The second mirror is configured to guide the fluorescence emission beam from the first mirror to the third mirror, and the third mirror is configured to guide the fluorescence emission beam into the camera. In some embodiments, a first optical filter is located between the beam splitter and the first mirror, and a second optical filter is located between the third mirror and the camera.

[0007] In some embodiments, the second arm further includes a focusing mechanism configured to focus the fluorescence emission beam received by the objective lens from each specimen. In some embodiments, the light source is supported by the second arm.

[0008] The drive system is configured to move the objective lens in the XY plane below the stage. A controller is coupled to the drive system and configured to control the drive system to selectively position the objective lens below each window in the array. In some embodiments, the drive system includes a first drive assembly configured to move the objective lens along the X direction in the plane, and a second drive assembly configured to move the objective lens along the Y direction in the plane. The controller is coupled to the light source and configured to control the operation of the light source.

[0009] The camera is located within the base and is configured to generate a fluorescence image of each specimen based on the corresponding fluorescence emission beam received from each specimen.

[0010] In some embodiments, the glass stage is movably fixed to the housing and can move between an open position and a closed position. In some embodiments, the glass stage comprises tempered glass, such as, for example, Gorilla® glass.

[0011] In some embodiments, the light source includes at least one solid-state light source.

[0012] The embodiments of the present invention offer advantages over conventional microscopes used in incubator environments because the optical components and other parts of the microscope are sealed and isolated from the incubator environment and are not exposed to pathogens, fungi, bacteria, moisture, etc., present therein. Furthermore, the microscope of the present invention uses objective lenses that scan beneath the specimen and has a much smaller footprint than conventional incubator microscopes. Additionally, the stage remains stationary, which is advantageous when there is no cell adhesion on the specimen plate being imaged.

[0013] It should be noted that aspects of the invention described with respect to one embodiment may be combined in different embodiments, although not specifically described with respect to it. That is, features of all embodiments and / or any embodiment may be combined in any manner and / or combination. The applicant reserves the right to accordingly amend any originally filed claim or to file any new claim, including the right to modify any originally filed claim to reference and / or incorporate any feature of any other claim, although not initially claimed in this manner. These and other objects and / or aspects of the invention are explained in detail below. Attached Figure Description

[0014] The accompanying drawings, which form part of this specification, illustrate various embodiments of the invention. Together with the description, the drawings serve to fully explain embodiments of the inventive concept.

[0015] Figure 1 This is a top perspective view of a fluorescence microscope according to some embodiments of the concept of the present invention.

[0016] Figure 2A and Figure 2B The diagram shows Figure 1 The microscope has a transparent housing, allowing the optical components and other microscope parts inside to be seen. Figure 2A In the middle, the glass stage on which the sample is placed for scanning and imaging is in the open position. Figure 2B In the middle, the glass stage is in a closed position, thus sealing and isolating the inside of the shell from the external environment.

[0017] Figure 3 yes Figure 1 A perspective view of the microscope, in which the shell has been removed for clarity.

[0018] Figure 4A It is a section taken along line 4A-4A. Figure 3 A side view of the microscope.

[0019] Figure 4B It is cut along line 4B-4B. Figure 3 A side view of the microscope.

[0020] Figure 5A and Figure 5B yes Figure 3 A side view of the optical components of the microscope, taken along line 5-5, and for clarity of other components of the microscope not shown.

[0021] Figure 6 This is a top plan view of the glass stage with the optical components below. Other components of the microscope, not shown, are for clarity. Detailed Implementation

[0022] Original Reference Figure 1 and Figures 2A-2B The illustration shows a fluorescence microscope 10 for use in an incubator environment according to an embodiment of the present invention. The microscope 10 includes a housing 20 having a generally cubic shape, comprising upper and lower walls 21, 22, and side walls 23, 24, 25, 26. The upper wall 21 includes an opening 21a through which the optical components and other parts of the microscope 10 are visible. A glass stage 30 is movably fixed to the housing 20 via a pair of hinges 32 and can be in an open position (…). Figure 1 and Figure 2A ) and closed position ( Figure 2B The stage 30 moves between the opening 21a in the upper wall 21 of the housing 20. When in the closed position, the glass stage 30 covers the opening 21a in the upper wall 21 of the housing and seals the interior of the housing 20 from the external environment, such as an incubator environment. The housing 20 can be formed from a variety of materials, including but not limited to metals (e.g., stainless steel) and polymers, and combinations thereof. It will be appreciated by those skilled in the art that any number of suitable materials can be used, as long as they can withstand the environment inside the incubator. For example, for human cells, incubators are typically maintained at a temperature of around 37°C, although other higher or lower temperatures can be utilized for any number of specimens. Furthermore, the atmosphere inside the incubator is frequently humidified and enriched with CO2, for example, to prevent live cells from drying out. In some incubators, oxygen in the atmosphere is limited. Therefore, any material that will not be adversely affected by such an environment can be used to form the housing 20 of the microscope 10. Furthermore, the housing 20 can have a variety of shapes and configurations and is not limited to the shape illustrated. In at least some embodiments, the housing 20 is a box that is substantially rectangular or square, but those skilled in the art will recognize that any number of shapes can be suitable. The microscope 10 can be used in any incubator, as long as the incubator is physically able to accommodate the size of the microscope 10. The advantage of the bi-arm configuration of the fluorescence microscope 10 described below is that it allows the fluorescence microscope 10 to be small enough to fit into a standard incubator.

[0023] The illustrated glass stage 30 includes a frame 34 supporting an array of windows 36. In at least some embodiments, the frame 34 is made of stainless steel. The frame 34 can be formed from a variety of materials, including but not limited to metals and polymers, and combinations thereof. As those skilled in the art will understand, the dimensions of each window 36 are set and each window 36 is configured to receive a corresponding specimen plate. In the illustrated embodiment, the glass stage includes six windows 36. However, the glass stage 30 may include a variety of numbers of windows 36, including more than six windows and fewer than six windows, and embodiments of the inventive concept are not limited to the number and arrangement of the windows 36 illustrated. In some embodiments, the windows 36 of the glass stage 30 are formed from tempered glass, such as, for example, Gorilla® glass (available from Corning, New York), and in some embodiments, the windows 36 of the glass stage 30 are formed from Dragontail™ glass (available from Asahi Glass, Tokyo, Japan) or Xensation® glass (available from Schott AG, Mainz, Germany).

[0024] refer to Figure 3 and Figures 4A-4B The illustration shows various components of the microscope 10 located within the housing 20, with the housing 20 removed for clarity. These components include a camera 100, a light source 200, an optical assembly 300, and a drive system 400. The optical assembly 300 is movable, as will be described below, to provide an optical path for an illumination beam from the light source 200 to a specimen positioned at each window 36 of the stage 30, and to provide an optical path for a fluorescence emission beam from each specimen to the camera 100. The optical assembly 300 is configured such that the length of the optical path for the fluorescence emission beam from each specimen to the camera 100 is substantially the same. Therefore, regardless of which window 36 the objective lens 330 is positioned under, the optical path for the fluorescence emission beam from the specimen will have the same length. In some embodiments, the objective lens 330 may include lenses from 1.25x to 40x, but those skilled in the art will recognize that any objective intensity can be used depending on the working distance and the specimen to be viewed and / or imaged.

[0025] refer to Figures 5A-5BThe illustration shows an optical assembly 300, with other components of the microscope 10 removed for clarity. The optical assembly 300 includes a base 302 fixed to the lower wall 22 of the housing 20. In the illustrated embodiment, the base 302 includes a generally cylindrical housing 304 that houses the camera 100. The lower portion of the base housing 304 is fixed to a plate 306, which is also fixed to the lower wall 22 of the housing 20. The optical assembly 300 includes a first arm 310 movably fixed to the upper portion of the base housing 304 via a bearing 308 and rotatable about a first axis A1 via the bearing 308. The bearing 308 is annular in shape and has a central opening that allows an optical path of fluorescence emission beams from each specimen to reach the camera 100 located within the base housing 304.

[0026] The optical assembly 300 also includes a second arm 320 movably fixed to the first arm 310 via a bearing 322 and rotatable about a second axis A2, which is spaced apart from and substantially parallel to the first axis A1, as illustrated. The bearing 322 is annular in shape and has a central opening that allows an optical path of fluorescence emission beams from each specimen from the second arm 320 into the first arm 310.

[0027] The first arm 310 and the second arm 320 each include optical components that generate an optical path for a fluorescence emission beam from each specimen to the camera 100. In the illustrated embodiment, the second arm 320 includes an objective lens 330 configured to move in a plane below the stage 30 as the first and second arms 310, 320 rotate about their respective axes A1, A2. The second arm 320 also includes a light source 200 for illuminating the sample supported by the stage 30. In some embodiments, the light source includes at least one solid-state light source, such as a light-emitting diode (LED) or a semiconductor laser. Light from the light source 200 passes through the objective lens 330 and onto the specimen located at a corresponding window 36 above the objective lens 330. The fluorescence emission beam from the sample then returns through the objective lens 330, passes through the first and second arms 310, 320, and onto the camera 100. The camera 100 is configured to generate a fluorescence image of each specimen based on the corresponding fluorescence emission beam received from each specimen, as will be understood by those skilled in the art. Camera 100 may be, for example, a charge-coupled device (CCD) camera, a complementary metal-oxide-semiconductor (CMOS) camera, or a scientific CMOS (sCMOS) camera.

[0028] In the illustrated embodiment, the second arm 320 includes a focusing mechanism 332 configured to focus the fluorescence emission beam received by the objective lens 330 from each specimen. The focusing mechanism 332 may be a deformable lens capable of adjusting the focus in the Z-direction (i.e., the direction perpendicular to the XY plane). In some embodiments, the deformable lens may be water-based or gel-based. In some embodiments, the deformable lens may be manipulated or deformed by an electric field, piezoelectric deformation, or a circular actuator.

[0029] The drive system 400 is configured to move the objective lens in the XY plane below the stage 30. Specifically, the drive system 400 is configured to move the first and second arms 310, 320 of the optical assembly 300 such that the objective lens 330 can scan back and forth in the XY plane below each window 36 in the array. (Reference) Figure 6 The diagram illustrates an array of windows 36 with optical components 300 positioned below them. The drive system 400 is configured to move the optical components (i.e., to rotate the first and second arms 310, 320 about their respective axes A1, A2) such that the objective lens 330 can be positioned at each window 36 in the array and can scan back and forth in the XY plane below each respective window 36, thereby acquiring an image of the specimen in the corresponding window before moving to another window. For example, the objective lens can initially be positioned below window 36a in the upper left portion of the array and then moved back and forth to scan the entire or other predefined area of ​​window 36a to acquire an image of the sample positioned at window 36a. The objective lens can then be moved to the adjacent window 36b and the same scanning operation can be performed to acquire an image of the sample at window 36b. This continues until all windows 36a-36f (or at least all windows on which samples are placed) and the samples supported by them have been imaged.

[0030] Go back to reference Figure 3The illustrated drive system 400 includes a first drive assembly 410 configured to move an objective lens 330 along the X direction in the XY plane, and a second drive assembly 420 configured to move the objective lens along the Y direction in the XY plane. The first drive assembly includes a drive motor 412 having an output shaft coupled to a drive pulley 414. A drive belt 416 is driven by the drive pulley 414 to move the objective lens 330 along the X direction in the XY plane. The drive belt 416 surrounds a series of pulleys 418, as those skilled in the art will understand. Similarly, the second drive assembly 420 includes a drive motor 422 having an output shaft coupled to a drive pulley 424. A drive belt 426 is driven by the drive pulley 424 to move the objective lens 330 along the Y direction in the XY plane. The drive belt 426 surrounds a series of pulleys 428, as those skilled in the art will understand. The drive motors 412 and 422 are operated by one or more controllers 450. Figure 3 and Figures 4A-4B The one or more controllers 450 can be located anywhere in the open space of the instrument without restriction. In some embodiments, the one or more controllers 450 may be located outside the housing 20. Power to the drive motors 412, 422 and to the light source 200 and camera 100 is supplied from a power source via power cables 500.

[0031] Embodiments of the present invention are not limited to the illustrated drive system 400. For example, in other embodiments, the drive system 400 may utilize any of a voice coil, a linear motor, a stepper motor per axis, or an in-core XY system to move the first and second arms 310, 320, thereby selectively positioning the objective lens 330 under the stage 30.

[0032] In the illustrated embodiment, the drive system 400 is supported within the housing 20 by a frame 430. The illustrated frame 430 includes four adjustable vertical supports 440 that support a pair of elongated members 442. The elongated members 442 support drive motors 412, 422, and various pulleys 418, 428 around which corresponding drive belts 416, 426 engage. Each of the illustrated vertical supports 440 has a lower portion 440a and an upper portion 440b, the upper portion 440b being telescopically movable relative to the lower portion 440a such that the height of the elongated members 442 supporting the drive system 400 can be adjusted relative to the lower wall 21 of the housing 20. A locking mechanism 444 is provided to each support 440 to hold the upper portion 440b at a desired height. Furthermore, in the illustrated embodiment, the four adjustable vertical supports 441 hold a top plate and a glass sealing plate with openings for the MTP. In other embodiments, the height difference may be fixed and the vertical supports 440, 441 may be replaced by a fixed-height support system.

[0033] Go back to reference Figures 5A-5B In addition to supporting the objective lens 330, the second arm 320 also includes a beam splitter 322 and a first mirror 326. The beam splitter 322 is configured to allow an illumination beam from the light source 200 to pass through it to the objective lens 330 and onto the specimen at a corresponding window 36 of the stage 30. The beam splitter 322 is also configured to guide a fluorescence emission beam received by the objective lens 330 from the corresponding specimen through the first arm 310 to the first mirror 326. In at least some embodiments, the beam splitter 322 is a dielectric-coated glass and / or quartz substrate. In some embodiments, the beam splitter 322 may be a 50 / 50 beam splitter, but those skilled in the art will recognize the benefits of using a beam splitter with a dielectric-coated glass / quartz substrate, including improved transmission and reflection.

[0034] The first arm 310 includes a second mirror 312 and a third mirror 314. The second mirror 312 is configured to guide a fluorescence emission beam from the first mirror 326 to the third mirror 314. The third mirror 314 is configured to guide the fluorescence emission beam into the camera 100. In some embodiments, an optical filter 324 is located between the beam splitter 322 and the first mirror 326. In some embodiments, the optical filter may be located between the third mirror 314 and the camera 100 to limit detection to the desired spectrum of the sample. Embodiments of this concept can also be used to detect specimens in transmitted light mode. In this case, the optical filter allows the transmitted light source to pass through and blocks ambient light. In fluorescence mode, the filter is ideally positioned in the pupil conjugant plane because dust on it is subsequently invisible in the image. However, they can be inserted somewhere in the detection beam path after the beam splitter 322 to enable the instrument to function. Optimal background reduction is achieved directly in front of the camera, and is most user-friendly in a filter block containing the light source 200, the beam splitter 322, and the optical filter 324. In some embodiments, deep learning or artificial intelligence (AI) can be used to detect specimens. In some such embodiments, a planar transmitted light source can be placed on top of a microtiter plate and can be activated (i.e., turned on) (if objective 320 is in place and instrument 10 is ready for image acquisition). Instead of showing markers like fluorescence contrast in the transmitted light image, the AI ​​can be trained using deep learning algorithms to detect cells or different compartments.

[0035] Figure 5B The diagram illustrates the optical path P1 of light from the light source 200 through the objective lens 330 to the specimen at the corresponding window 36 of the stage 30. Figure 5BThe optical path P2 of the fluorescence emission beam from the corresponding specimen to the camera 100 is also illustrated. The length L of the optical path P2 is the sum of the following distances: the distance from the imaged specimen to the beam splitter 322, the distance from the beam splitter 322 to the first mirror 326, the distance from the first mirror 326 to the second mirror 312, the distance from the second mirror 312 to the third mirror 314, and the distance from the third mirror 314 to the camera 100. Due to the movable configuration of the first and second arms 310, 320, the length L of the optical path P2 will be substantially the same regardless of which window 36 the objective lens 330 is positioned under. The advantage of this dual-arm configuration is that it combines low weight (meaning that the optical assembly 300 can move the objective lens 330 very quickly under the sample area) and stable optical performance due to always maintaining the same optical path length. However, those skilled in the art will recognize that any number of arms can be suitable. For example, the first arm 310 or the second arm 320 may comprise one or more arms for the same purpose. In some embodiments, there are a first arm 310, a second arm 320, and a third arm (not shown), the third arm also including two additional mirrors for substantially the same purpose as those on the second arm 320. For good optical quality in modern objective lenses, the distance between the tube lens (between mirrors 326 and 312) and the camera 100 must be constant (otherwise the magnification would change). As those skilled in the art will recognize, the distance from the rear aperture of objective lens 330 to the tube lens can vary slightly (typically with a focus drive present), but if the variation exceeds approximately + / - 15 mm, image distortion and vignetting may occur.

[0036] The controller 450 controls all moving parts and data collection parts of the microscope 10. An image-based autofocus algorithm allows the controller 450 to turn on the light source and capture an image, then change the focal plane using a tunable lens and capture another image, and so on. The algorithm then determines the image to be in focus from the remaining images.

[0037] In some embodiments, the controller 450 may include a processor, which may take the form of suitable software and a microprocessor and / or computer for controlling microscope functions and analyzing any samples and / or specimens viewed and / or imaged by the microscope. In some embodiments, the processor may be present in, for example, the controller 450 or as a separate processor to control and coordinate system operation for various operating modes of using the system. For this purpose, the processor may be electrically coupled to each component of the system (e.g., objectives, scanning stage, light source, one or more cameras, one or more drive systems, etc.). In some aspects, as those skilled in the art will recognize, the microscope 10 may include additional elements that can be controlled by the controller 450 and / or the processor, such as, but not limited to, a filter changer and a barcode scanner for identifying the current objective and an excitation module.

[0038] In some configurations, controller 450 may include, for example, shared hardware and / or one or more computer systems having suitable software and / or a microprocessor for operating the system (e.g., for controlling drive systems, drive motors, light sources, cameras(s), etc.). In some examples, any one or more components of the system may include their respective controllers, processors, operating systems, and other features to allow operation of that component. Controller 450 may be integrated with the system or may reside on one or more accessory boards, printed circuit boards, or computers electrically coupled to components of the system. Controller 450 is typically electrically coupled to one or more memory cells to receive data from other components of the system and to allow adjustment of various system parameters as needed or desired. The processor may be part of a general-purpose computer, such as Unix-based processors, Intel Pentium processors, ARM processors, Motorola PowerPC, Sun UltraSPARC, Hewlett-Packard PA-RISC processors, or any other type of processor. Depending on various embodiments of the technology, one or more of any type of controller or computer system may be used. Additionally, the system or controller 450 may be connected to a single computer or may be distributed among multiple computers attached via a communication network. It should be recognized that other functions (including network communication) can be performed, and the technology is not limited to having any particular function or set of functions. Various aspects can be implemented as specialized software executing in a general-purpose computer system. A computer system may include a processor connected to one or more storage devices (such as disk drives, memory, or other devices for storing data). Memory is typically used to store data, calibrations, and programs during system operation in various modes. Components of a computer system can be coupled via interconnect devices, which may include one or more buses (e.g., between components integrated within the same machine) and / or networks (e.g., between components located on separate, discrete machines). Interconnect devices are provided for communication (e.g., signals, data, instructions) to be exchanged between components of the system. A computer system can typically receive and / or issue commands within processing time (e.g., milliseconds, microseconds, or less) to allow for rapid control of the system. Controller 450 and / or processor are typically electrically coupled to a power source, which may be, for example, a DC power source, an AC power source, a battery, a fuel cell, or other power source or a combination of power sources. The power source may be shared by other components of the system. The system may also include one or more input devices (e.g., keyboard, mouse, trackball, microphone, touch screen, manual switch (e.g., over-the-air switch)) and one or more output devices (e.g., printer, display screen, speaker).In addition, the system may include one or more communication interfaces that connect the computer system and / or controller 450 to a communication network (as a supplement to or replacement of interconnected devices). The system may also include suitable circuitry to convert signals received from various electrical devices present in the system. Such circuitry may be present on a printed circuit board or on a separate board or device electrically coupled to the printed circuit board via a suitable interface (e.g., a serial ATA interface, an ISA interface, a PCI interface, etc.) or via one or more wireless interfaces (e.g., Bluetooth®, Wi-Fi®, near-field communication, or other wireless protocols and / or interfaces).

[0039] In some embodiments, the storage system used in the systems described herein typically includes a computer-readable and writable non-volatile recording medium in which code that can be used by a program executable by a processor is stored, or information is stored on or in the medium for processing by a program. The medium can be, for example, a hard disk, a solid-state drive, or flash memory. Typically, in operation, the processor causes data to be read from the non-volatile recording medium and into another memory that allows the processor to access information faster than the recording medium. This memory is typically volatile random access memory, such as dynamic random access memory (DRAM) or static random access memory (SRAM). It can reside in a storage system or memory system. The processor generally manipulates the data within the integrated circuit memory and then copies the data to the medium after processing is complete. Various mechanisms are known for managing data movement between the medium and integrated circuit memory elements, and the technology is not limited thereto. The technology is also not limited to a particular memory system or storage system. In some embodiments, the system may also include dedicated hardware that is specifically programmed. Aspects of the technology can be implemented in software, hardware, or firmware, or any combination thereof. Furthermore, such methods, actions, systems, system elements, and components can be implemented as part of the aforementioned systems or as independent components. While specific systems are described by way of example as a class of systems on which various aspects of the technology can be practiced, it should be understood that these aspects are not limited to implementation on the described systems. The aspects can be practiced on one or more systems with different architectures or components. Systems can include general-purpose computer systems that can be programmed using high-level computer programming languages. Systems can also be implemented using specialized hardware that is specifically programmed. Processors can be commercially available processors, such as the well-known Pentium-class processors available from Intel. Many other processors are also commercially available. Such processors typically run operating systems, which can be, for example, Windows 95, Windows 98, Windows NT, Windows 2000 (Windows ME), Windows XP, Windows Vista, Windows 7, Windows 8, or Windows 10 operating systems available from Microsoft, MAC OSX, such as Snow Leopard, Lion, Mountain Lion, or other versions available from Apple, Solaris operating systems available from Sun Microsystems, or UNIX or Linux operating systems available from various sources. Many other operating systems can be used, and in some embodiments, a simple set of commands or instructions can serve as the operating system.

[0040] In some examples, the processor and operating system may together define a platform for which applications can be written using a high-level programming language. It should be understood that the technology is not limited to a specific system platform, processor, operating system, or network. Furthermore, it will be apparent to those skilled in the art that, having benefited from this disclosure, the technology is not limited to a specific programming language or computer system. Additionally, it should be recognized that other suitable programming languages ​​and other suitable systems may also be used. In some examples, the hardware or software may be configured to implement a cognitive architecture, neural network, or other suitable implementation. If desired, one or more portions of the computer system may be distributed across one or more computer systems coupled to a communication network. These computer systems may also be general-purpose computer systems. For example, aspects may be distributed across one or more computer systems configured to provide services (e.g., servers) to one or more client computers, or to perform an overall task as part of a distributed system. For example, aspects may be implemented on a client-server or multi-tiered system comprising components distributed across one or more server systems that perform various functions according to various embodiments. These components can be executable code, intermediate code (e.g., IL), or interpreted code (e.g., Java), communicating over a communication network (e.g., the Internet) using a communication protocol (e.g., TCP / IP). It should also be recognized that the technology is not limited to execution on any particular system or group of systems. Furthermore, it should be recognized that the technology is not limited to any particular distributed architecture, network, or communication protocol.

[0041] In some instances, various embodiments can be programmed using object-oriented programming languages ​​such as, for example, SQL, Smalltalk, Basic, Java, Javascript, PHP, C++, Ada, Python, iOS / Swift, Ruby on Rails, or C# (C-Sharp) . Other object-oriented programming languages ​​may also be used. Alternatively, functional, scripting, and / or logic programming languages ​​may be used. Various configurations can be implemented in non-programming environments (e.g., documents created in HTML, XML, or other formats that, when viewed in a browser program window, render aspects of a graphical user interface (GUI) or perform other functions). Some configurations can be implemented as programmable or non-programmable elements or any combination thereof. In some instances, the system may include remote interfaces, such as those present on mobile devices, tablets, laptops, or other portable devices, which can communicate via wired or wireless interfaces and allow remote operation of the system as desired.

[0042] Embodiments of the invention have been described above with reference to the accompanying drawings, which illustrate embodiments of the invention. However, the invention can be embodied in many different forms and should not be construed as limited to the embodiments set forth herein. As will be appreciated by those skilled in the art based on the teachings herein, numerous changes and modifications can be made to the above and other embodiments without departing from the scope defined in the claims. Rather, these embodiments are provided to make this disclosure thorough and complete and to fully convey the scope of the invention to those skilled in the art. For example, components of the microscope 10, housing 20, frame 34, or stage 30 can be made of any of many different materials now known or hereafter known. For example, but not limited to, the stage can be made of suitable plastic or glass alternatives such as quartz, or the stage can in other cases comprise a single window for imaging a sample or specimen, rather than taking the form of a stage with a frame defining multiple windows. Accordingly, this detailed description of the embodiments should be taken in an illustrative rather than restrictive sense. Similar reference numerals throughout the drawings refer to similar elements.

[0043] It will be understood that while the terms first, second, etc., may be used herein to describe various elements, these elements should not be limited by these terms. These terms are used only to distinguish one element from another. For example, a first element may be referred to as a second element, and similarly, a second element may be referred to as a first element, without departing from the scope of the invention. As used herein, the term "and / or" includes any and all combinations of one or more of the associated listed items.

[0044] It will be understood that when an element is referred to as being "on" another element, it can be directly on that element or there may be an intermediate element present. In contrast, when an element is referred to as being "directly on" another element, there is no intermediate element present. It will also be understood that when an element is referred to as being "connected" or "coupled" to another element, it can be directly connected or coupled to that element or there may be an intermediate element present. In contrast, when an element is referred to as being "directly connected" or "directly coupled" to another element, there is no intermediate element present. Other terms used to describe relationships between elements should be interpreted in a similar manner (i.e., "between" versus "directly between," "adjacent to" versus "directly adjacent to," etc.).

[0045] Relative terms such as “below” or “above” or “up” or “down” or “horizontal” or “vertical” may be used herein to describe the relationship between one element, layer, or region and another element, layer, or region as shown in the figures. It will be understood that these terms are intended to cover different orientations of the device in addition to those shown in the figures.

[0046] The terminology used herein is for the purpose of describing particular embodiments only and is not intended to limit the invention. As used herein, the singular forms “a” and “the” are intended to include the plural forms as well, unless the context clearly indicates otherwise. It will be further understood that, when used herein, the terms “comprises,” “comprising,” “includes,” and / or “including” specify the presence of the stated features, operations, elements, and / or components, but do not preclude the presence or addition of one or more other features, operations, elements, components, and / or groups thereof.

[0047] All aspects and elements of the embodiments disclosed above can be combined in any way and / or in combination with aspects or elements of other embodiments to provide multiple additional embodiments.

Claims

1. A fluorescence microscope, comprising: A housing, the housing including an upper wall having an opening; A glass stage with an opening at the top, wherein the glass stage seals and isolates the interior of the housing from the external environment and includes an array of windows, each window being configured to receive a specimen plate; The camera is located inside the housing; A light source located within the housing, wherein the light source is configured to generate an illumination beam; as well as An optical assembly located within the housing, wherein the optical assembly is movable to provide an optical path from a light source to an illumination beam positioned at each window in the array, and to provide an optical path from each specimen to a fluorescence emission beam from the camera, wherein the length of the optical path from each specimen to the camera is substantially the same.

2. The fluorescence microscope of claim 1, wherein the optical components include: A base fixed to the lower wall of the housing; A first arm is movably fixed to the base, wherein the first arm is configured to rotate about a first axis; as well as A second arm is movably fixed to the first arm, wherein the second arm is configured to rotate about a second axis that is spaced apart from and substantially parallel to the first axis; The first and second arms each include optical components that generate an optical path for the fluorescence emission beam from each specimen to the camera.

3. The fluorescence microscope of claim 2, wherein the second arm includes an objective lens, and the fluorescence microscope further includes a drive system configured to move the objective lens in a plane below the stage and a controller coupled to the drive system, wherein the controller is configured to control the drive system to selectively position the objective lens below each window in the array.

4. The fluorescence microscope of claim 3, wherein the driving system includes a first driving assembly configured to move the objective lens along the X direction in the plane and a second driving assembly configured to move the objective lens along the Y direction in the plane.

5. The fluorescence microscope of claim 3, wherein the optical components of the second arm include a beam splitter and a first mirror, wherein the beam splitter is configured to guide an illumination beam from a light source through the objective lens and onto the corresponding specimen and to guide a fluorescence emission beam received by the objective lens from the corresponding specimen to the first mirror, and wherein the first mirror is configured to guide the fluorescence emission beam into the first arm.

6. The fluorescence microscope of claim 5, wherein the optical components of the first arm include a second mirror and a third mirror, wherein the second mirror is configured to guide a fluorescence emission beam from the first mirror to the third mirror, and wherein the third mirror is configured to guide the fluorescence emission beam into a camera.

7. The fluorescence microscope of claim 6 further includes a first optical filter located between the beam splitter and the first mirror, and a second optical filter located between the third mirror and the camera.

8. The fluorescence microscope of claim 3, wherein the second arm further includes a focusing mechanism configured to focus the fluorescence emission beam received by the objective lens from each specimen.

9. The fluorescence microscope of claim 2, wherein the light source is supported by the second arm.

10. The fluorescence microscope of claim 3, wherein the controller is coupled to the light source, and wherein the controller is further configured to control the operation of the light source.

11. The fluorescence microscope of claim 2, wherein the camera is located within the base and configured to generate a fluorescence image of each specimen based on a corresponding fluorescence emission beam received from each specimen.

12. The fluorescence microscope of claim 1, wherein the glass stage is movably fixed to the housing and is movable between an open position and a closed position.

13. The fluorescence microscope of claim 1, wherein the stage comprises tempered glass.

14. The fluorescence microscope of claim 13, wherein the tempered glass is Gorilla® glass.

15. The fluorescence microscope of claim 1, wherein the light source comprises at least one solid-state light source.

16. A fluorescence microscope, comprising: A housing, the housing including an upper wall having an opening; A glass stage with an opening at the top, wherein the glass stage seals and isolates the interior of the housing from the external environment and includes an array of windows, each window being configured to receive a specimen plate; The camera is located inside the housing; A light source located within the housing, wherein the light source is configured to generate an illumination beam; An optical assembly located within a housing, comprising an objective lens, and wherein the optical assembly is movable to provide an optical path for an illumination beam from a light source through the objective lens to a specimen positioned at each window in the array, and to provide an optical path for a fluorescence emission beam from each specimen through the objective lens to a camera, wherein the length of the optical path from each specimen to the camera is substantially the same. as well as A drive system configured to move the objective lens in a plane below the glass stage and to selectively position the objective lens below each window in the array.

17. The fluorescence microscope of claim 16, wherein the driving system includes a first driving assembly configured to move the objective lens along the X direction in the plane, and a second driving assembly configured to move the objective lens along the Y direction in the plane.

18. The fluorescence microscope of claim 16, wherein the glass stage is movably fixed to the housing and movable between an open position and a closed position, and wherein the glass stage comprises Gorilla® glass.

19. A fluorescence microscope, comprising: A housing, the housing including an upper wall having an opening; A glass stage with an opening at the top, wherein the glass stage seals and isolates the interior of the housing from the external environment, wherein the glass stage includes an array of windows, each window being configured to receive a specimen plate; The camera is located inside the housing; as well as An optical component located within the housing, the optical component comprising: A base fixed to the lower wall of the housing; A first arm movably fixed to the base, wherein the first arm is configured to rotate about a first axis; and A second arm is movably fixed to the first arm, wherein the second arm is configured to rotate about a second axis that is spaced apart from and substantially parallel to the first axis; The second arm includes an objective lens, a light source configured to generate an illumination beam, a beam splitter, and a first mirror, wherein the beam splitter is configured to guide the illumination beam from the light source through the objective lens and onto the specimen positioned at a corresponding window, and to guide the fluorescence emission beam from the specimen received by the objective lens to the first mirror, and wherein the first mirror is configured to guide the fluorescence emission beam into the first arm. The first arm includes a second mirror and a third mirror, wherein the second mirror is configured to guide the fluorescence emission beam from the first mirror to the third mirror, and wherein the third mirror is configured to guide the fluorescence emission beam into the camera; The first and second arms are movable, ensuring that the optical path length of the fluorescence emission beam from each specimen to the camera is substantially the same.

20. The fluorescence microscope of claim 19, further comprising a drive system configured to move the objective lens in a plane below the stage and selectively position the objective lens below each window in the array, wherein the drive system includes a first drive assembly configured to move the objective lens in the plane along an X-direction and a second drive assembly configured to move the objective lens in the plane along a Y-direction.

21. The fluorescence microscope of claim 19, wherein the stage comprises tempered glass.

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