Enclosed confocal optical system

Abstract

An optical instrument includes a housing including a window and a port configured to receive light. The instrument includes an objective lens configured to align with the window, and an optical module located inside the housing. The optical module includes a dichroic mirror and one or more first optical elements configured to direct light from the port toward the dichroic mirror. The optical module includes one or more second optical elements configured to direct light from the dichroic mirror through the window to the objective lens, and configured to direct light from the window toward the dichroic mirror. The instrument includes a plate having openings patterned for confocal microscopy. The plate is configured to be inserted into, or removed from, a region between the dichroic mirror and the second optical elements. The optical instrument includes an image sensor and a third optical element configured to direct light from the dichroic mirror toward the image sensor.

Description

[0001] This application claims priority to U.S. Patent Application No. 19 / 187,156, filed April 23, 2025. Technical Field

[0002] This invention relates generally to the field of optical imaging, and more specifically to an optical instrument and its operating method. Background Technology

[0003] Researchers in many scientific fields value information that can be obtained through imaging live cells, such as by detecting and quantifying cellular fluorescence emission under physiologically relevant environmental conditions over extended periods. Some imaging systems include small environmental chambers attached to the stage of a small microscope. However, the environmental control capabilities of such chambers are limited, thus restricting the types of experiments that can be performed on cells. Maintaining precise target temperature and humidity for large quantities of biological samples over extended periods, while ensuring the availability of biosamples for analysis, remains challenging. Summary of the Invention

[0004] A first example is an optical instrument comprising: a housing including: a window; and a port configured to receive first light via an optical fiber; an objective lens configured to be aligned with the window such that the first light emitted from the housing through the window enters the objective lens, and second light from the objective lens enters the housing through the window; and an optical module located inside the housing, the optical module including: a dichroic mirror; one or more first optical elements configured to direct the first light from the port to the dichroic mirror; one or more second optical elements configured to guide the first light from the dichroic mirror through the window to the objective lens, and the one or more second optical elements configured to direct the second light from the window to the dichroic mirror; a plate having an opening patterned for rotary confocal microscopy, wherein the plate is configured to be inserted into or removed from a region between the dichroic mirror and the one or more second optical elements; an image sensor; and one or more third optical elements configured to direct the second light from the dichroic mirror to the image sensor.

[0005] A second example is a method of operating an optical instrument, the method comprising: moving an optical module relative to a stage supporting a first biological sample and a second biological sample, thereby aligning the optical module with the first biological sample, wherein the optical module includes a plate having an opening patterned for rotary confocal microscopy; capturing one or more first images of the first biological sample using the optical module; moving the optical module relative to the stage, thereby aligning the optical module with the second biological sample; and capturing one or more second images of the second biological sample using the optical module.

[0006] The third example is a non-transitory computer-readable medium that stores instructions that, when executed by the optical instrument described in the first example, cause the optical instrument to perform the method described in the second example.

[0007] When the terms “substantially” or “about” are used herein, it means that the feature, parameter, or value does not need to be precisely achieved, but may deviate or vary, including, for example, tolerances, measurement errors, measurement accuracy limitations, and other factors known to those skilled in the art, and that the amount of such deviation or variation does not impair the effect that the feature is intended to achieve. In some examples disclosed herein, “substantially” or “about” means within ±0-5% of the value.

[0008] These and other aspects, advantages, and alternatives will become clear to those skilled in the art upon reading the following detailed description and referring to the accompanying drawings where appropriate. Furthermore, it should be understood that the content of this invention, as well as the other descriptions and drawings provided herein, are intended to illustrate the invention by way of example only, and therefore various modifications are possible. Attached Figure Description

[0009] Figure 1 This is a block diagram of a computing device based on an example.

[0010] Figure 2 This is a block diagram of a computing device, light source, and optical instrument housed within an environmental chamber, based on an example.

[0011] Figure 3 This is a schematic diagram of an optical instrument, an environmental chamber, and a computing device, based on an example.

[0012] Figure 4 This is a schematic diagram of an optical instrument housed in a casing, based on an example.

[0013] Figure 5 This is a schematic diagram of an optical instrument after removing its casing, based on an example.

[0014] Figure 6 This is a schematic diagram of the exterior of the housing port according to the example.

[0015] Figure 7 This is a schematic diagram of the housing, selection module, and transmission light module according to the example.

[0016] Figure 8 This is another schematic diagram of the housing, selection module, and transmission light module based on the example.

[0017] Figure 9 This is a cross-sectional view of an optical module located inside the housing, based on an example.

[0018] Figure 10 This is a schematic diagram of the board based on the example.

[0019] Figure 11 This is a cross-sectional schematic diagram of a transmission light module based on an example.

[0020] Figure 12 This is a transparent view of the transmission light module based on the example.

[0021] Figure 13 This is a block diagram based on the example method. Detailed Implementation

[0022] As mentioned above, conventional instruments for live-cell imaging have several limitations. For example, many instruments can only culture a small number of cells and cannot perform several practical microsurgical modalities. The examples of optical instruments and their operating methods described herein can help alleviate these problems.

[0023] An optical instrument includes a housing comprising a window and a port configured to receive first light via an optical fiber. The first light is typically an excitation light in the form of a laser beam. The optical instrument also includes an objective lens configured to be aligned with the window such that first light exiting the housing through the window enters the objective lens, and second light from the objective lens enters the housing through the window. During use of the optical instrument, the objective lens is typically aligned with a sample such that second light originating from the sample is focused by the objective lens and passes through the window into the housing. The optical instrument includes an optical module located within the housing. The optical module includes a dichroic mirror configured to separate the first light (e.g., excitation light) from the second light (e.g., light originating from the sample). The optical module also includes one or more first optical elements (e.g., lenses, mirrors, prisms, filters, or gratings) configured to direct the first light from the port to the dichroic mirror. The optical module also includes one or more second optical elements configured to guide the first light from the dichroic mirror through the window into the objective lens (e.g., to focus the first light onto the sample). The one or more second optical elements are further configured to direct second light from the window to the dichroic mirror. The optical module also includes a plate (e.g., a disk) having an opening for patterning in a rotary confocal microscope. The plate is configured, depending on the operating mode, to be inserted into or removed from the region between the dichroic mirror and the one or more second optical elements. The optical module also includes an image sensor and one or more third optical elements configured to direct second light from the dichroic mirror to the image sensor.

[0024] As described in more detail below, the aforementioned optical instrument is configured to culture larger quantities of live cells for imaging analysis and is configured to perform various microscopy modalities, including rotary confocal microscopy. The optical module is typically sealed within a housing that isolates it from the external environmental conditions. This allows the optical module to be placed within a large environmental chamber, in close proximity to a large number of cultured biological samples. The enclosed optical module also facilitates the implementation of various microscopy modalities, such as rotary confocal microscopy, which are not feasible when using conventional microscopes and sample containment devices.

[0025] Confocal microscopy reduces defocus light, thus enabling optical tomographic imaging of three-dimensional biological structures. Rotary confocal microscopy, due to the parallel arrangement of openings (e.g., pinholes) within the rotary disk, achieves high acquisition speeds.

[0026] Figure 1This is a block diagram of a computing device 200. The computing device 200 includes one or more processors 202, a non-transitory computer-readable medium 204, a communication interface 206, and a user interface 208. The components of the computing device 200 can be connected together via a system bus, network, or other connection mechanism 212.

[0027] One or more processors 202 may be any type of processor, such as a microprocessor, a field-programmable gate array, a digital signal processor, a multi-core processor (such as a central processing unit (CPU) or a graphics processing unit (GPU)), etc., said processor being coupled to a non-transitory computer-readable medium 204.

[0028] The non-transitory computer-readable medium 204 can be any type of memory, such as volatile memory (e.g., random access memory (RAM), dynamic random access memory (DRAM), static random access memory (SRAM)), or non-volatile memory (e.g., read-only memory (ROM), flash memory, magnetic disk or optical disk, optical disc read-only memory (CD-ROM)), and other means for temporary or permanent storage of data or programs.

[0029] Furthermore, the non-transitory computer-readable medium 204 may store instructions 214. Instructions 214 may be executed by one or more processors 202 to cause the computing device 200 to perform any of the functions or methods described herein.

[0030] Communication interface 206 includes hardware capable of enabling communication within computing device 200 and / or communication between computing device 200 and one or more other devices. This hardware may include, for example, any type of input and / or output interface, Universal Serial Bus (USB), PCI Express, transmitter, receiver, and antenna. Communication interface 206 may be configured to facilitate communication with one or more other devices according to one or more wired or wireless communication protocols. For example, communication interface 206 may be configured to enable wireless data communication for computing device 200 according to one or more wireless communication standards, such as one or more IEEE 801.11 standards, ZigBee standards, Bluetooth standards, etc. As another example, communication interface 206 may be configured to enable wired data communication with one or more other devices. Communication interface 206 may also include an analog-to-digital converter (ADC) or a digital-to-analog converter (DAC), which computing device 200 may use to control various components of itself or external devices.

[0031] User interface 208 includes any type of display component configured to display data. As an example, user interface 208 may include a touchscreen display. As another example, user interface 208 may include a flat panel display, such as a liquid crystal display (LCD) or a light-emitting diode (LED) display. User interface 208 includes one or more hardware components for providing data and control signals to computing device 200. For example, user interface 208 may include a mouse or pointing device, a keyboard or keypad, a microphone, a touchpad or touchscreen, and other possible types of user input devices. Typically, user interface 208 enables an operator to interact with a graphical user interface (GUI) provided by computing device 200 (e.g., displayed by user interface 208).

[0032] Figure 2 This is a block diagram of a computing device 200, a light source 130A, and an optical instrument 100 housed within an environmental chamber 128. The computing device 200 is configured to control the optical instrument 100 and the light source 130A.

[0033] The light source 130A may be in the form of one or more lasers, one or more light-emitting diodes (LEDs), or one or more halogen lamps or arc lamps. The light source 130A is optically coupled to the optical instrument 100 via an optical cable 110 (e.g., a cable including one or more optical fibers). The light source 130A is typically located outside the ambient chamber 128.

[0034] The environmental chamber 128, also known as an incubator, is configured to maintain a first environmental condition that differs from the external second environmental condition. The environmental conditions controlled by the environmental chamber 128 may include one or more of temperature, humidity, or carbon dioxide partial pressure.

[0035] The optical instrument 100 includes a housing 102A, which houses an optical module 114. The optical instrument 100 also includes objective lenses 112A, 112B, and 112C, a stage 136 configured to hold one or more biological samples 20, and a transmission light module 138. The optical instrument 100 may also have... Figure 2 Showing fewer or more objectives.

[0036] Figure 3 This is a schematic diagram of the optical instrument 100, the environmental chamber 128, and the computing device 200. As shown, the stage 136 may be in the form of a pull-out drawer and is configured to hold one or more containers containing biological samples 20. The optical instrument 100, housed in the housing 102B, is configured to be placed within the environmental chamber 128.

[0037] Figure 4This is a schematic diagram of an optical instrument 100 housed in a housing 102B. The housing 102B includes a plurality of vents 160 that, when the optical instrument 100 and the housing 102B are placed in an environmental chamber 128, allow the biological sample 20 inside the housing 102B to be in the environmental conditions created by the environmental chamber 128.

[0038] Figure 5 This is a schematic diagram of the optical instrument 100 after removing the outer casing 102B.

[0039] The optical instrument 100 includes a housing 102A. The housing 102A may be sealed to maintain a lower humidity level inside than outside. (Reference) Figure 8 The optical module 114 within the housing 102A is at least partially isolated from the environmental conditions maintained by the environmental chamber 128. In some examples, the housing 102A is filled with an inert gas (such as nitrogen) or dehumidified air. The housing 102A also includes a window 104, which is made of, for example, glass or plastic. The housing 102A may be formed of one or more materials including acetal homopolymer resin, anodized aluminum, or stainless steel. The housing 102A may generally be made of one or more materials that possess resistance to moisture expansion, non-toxicity to cell culture, oxidation resistance, corrosion resistance, and / or thermal expansion resistance.

[0040] The optical instrument 100 also includes a frame 134 configured to support the housing 102B. The frame may be made of metal (such as stainless steel angle iron), but other examples may also be used. The frame 134 includes guide rails 137A and 137B.

[0041] The optical module 114 and housing 102A are configured to move relative to the stage 136 and frame 134 under the control of the computing device 200 and one or more motors. For example, housing 102A can move in a direction aligned with guide rail 137A and independently in a direction aligned with guide rail 137B. By moving relative to the stage 136, the optical module 114 can be used to illuminate and capture images of the biological sample 20 at different locations on the stage 136 without mechanically disturbing the biological sample 20. Housing 102A is slidably mounted on guide rail 137A, which is part of carriage 139. Carriage 139 and housing 102A can also move along guide rail 137B.

[0042] The optical instrument 100 also includes an objective lens selection module 132 (e.g., a rotatable turntable) operable to align objective lens 112A, objective lens 112B, or objective lens 112C with window 104.

[0043] The optical instrument 100 also includes a transmission light module 138, located outside the housing 102A and configured to emit a light beam toward the window 104. In typical operation of the transmission light module 138, the biological sample 20 and the stage 136 are positioned between the window 104 and the transmission light module 138. The transmission light module 138 is attached to a carriage 139 such that the transmission light module 138 moves synchronously with the housing 102A and remains aligned with the window 104.

[0044] Figure 6 This is an external schematic diagram of port 106 of housing 102A. Port 106 is configured to receive light 108A (e.g., a laser beam) via optical fiber 110. Therefore, port 106 is typically in the form of an optical coupler, configured to receive the end of optical fiber 110.

[0045] Figure 7 This is a schematic diagram of the housing 102A, the objective lens selection module 132, and the transmitted light module 138. The objective lens selection module 132 is operable (e.g., rotatable) to allow for the selection of objective lens 112A, objective lens 112B, or objective lens 112C for alignment with window 104. For example, when selected and properly arranged, objective lens 112A is aligned with window 104 such that light exiting housing 102A through window 104 enters objective lens 112A, and light from objective lens 112A enters housing 102A through window 104.

[0046] Figure 8 Another schematic diagram shows the housing 102A, the objective lens selection module 132, and the transmission light module 138. In typical operation of the transmission light module 138, the stage 136 and one or more biological samples 20 are located between the transmission light module 138 and the objective lens 112.

[0047] like Figure 8 As shown, objective lens 112A is aligned with window 104, so that light 108A emitted from housing 102A through window 104 enters objective lens 112A, and light 108B from objective lens 112B enters housing 102A through window 104.

[0048] The optical instrument 100 adjusts the depth of focus within the biological sample 20 by moving the objective lens selection module 132 and the objective lens 112 up and down relative to the stage 136. For example, the computing device 200 controls the motor 802 to move the shaft 804 up and down, thereby moving the objective lens selection module 132 and the objective lens 112 up and down. The shaft 804 is mechanically coupled to the objective lens selection module 132.

[0049] Figure 9This is a cross-sectional view of the optical module 114 located within the housing 102A. The optical module 114 includes a dichroic mirror 116A, a first optical element 118A, a second optical element 118B, a plate 120, an image sensor 126, and a third optical element 118C.

[0050] The dichroic mirror 116A can be used as an interference filter, configured to reflect light of a specific wavelength while transmitting light of other wavelengths. Figure 9 In the specific example shown, the dichroic mirror 116A is configured to transmit light 108A received from optical cable 110 via port 106 and reflect light 108B received from objective lens 112A. Port 106 is shown in... Figure 6 The 116A dichroic mirror is interchangeable with other dichroic mirrors that have different wavelength-dependent transmission or reflection characteristics.

[0051] A first optical element 118A is configured to guide light 108A from port 106 to a dichroic mirror 116A. The first optical element 118A includes a parabolic mirror 144 configured to guide light 108A received from port 106 to the dichroic mirror 116A via a rectangular microlens array 149. The first optical element 118A also includes the rectangular microlens array 149 configured to shape the light 108A received from the parabolic mirror 144. The first optical element 118A further includes a prism reflector 148A configured to reflect the light 108A received from the rectangular microlens array 149 to the dichroic mirror 116A. The first optical element 118A also includes a positive lens 150 configured to shape and guide the light 108A received from the prism reflector 148A to the dichroic mirror 116A. Dichroic mirror 116A is configured to transmit light 108A from positive lens 150 into region 121. Region 121 may or may not be occupied by plate 120.

[0052] The second optical element 118B is configured to guide light 108A from the dichroic mirror 116A through window 104 into the objective lens 112A, and to guide light 108B from window 104 into the dichroic mirror 116A. The second optical element 118B includes a prism mirror 148B configured to reflect light 108A from the dichroic mirror 116A to window 104, and to reflect light 108B back to the dichroic mirror 116A. The second optical element 118B also includes a barrel lens 152 configured to shape the light 108A from the prism mirror 148B and transmit the light 108A through window 104 to the objective lens 112A. The barrel lens 152 and the objective lens 112A are configured to focus light 108A onto the biological sample 20 held by the stage 136. The tube lens 152 is also configured to shape the light 108B from the objective lens 112A and transmit the light 108B to the prism mirror 148B.

[0053] refer to Figure 10 Plate 120 has an opening 131 patterned for rotary confocal microscopy. Plate 120 may be in the form of a glass disk, with the opening 131 formed by painting or metal patterning. The opening 131 may be in the form of pinholes arranged in one or more Archimedean spirals, or any other pinhole or slit arrangement within region 121, to allow scanning of an imaging area optically conjugate to image sensor 126. The opening 131 is configured to spatially filter light 108A and light 108B as plate 120 rotates within region 121.

[0054] refer to Figure 9 Plate 120 is configured to be inserted into or removed from region 121 between dichroic mirror 116A and second optical element 118B. Region 121 contains a conjugate focal plane relative to image sensor 126. When plate 120 is located within region 121, optical instrument 100 typically operates in confocal mode; when plate 120 is not located within region 121, optical instrument 100 operates in wide-field mode.

[0055] Image sensor 126 may be in the form of a complementary metal-oxide-semiconductor (CMOS) image sensor, but may also be in other example forms, such as a charge-coupled device (CCD), a scientific-grade CMOS (sCMOS) image sensor, or an electron multiplication charge-coupled device (EMCCD).

[0056] The third optical element 118C is configured to guide light 108B from the dichroic mirror 116A to the image sensor 126. The third optical element 118C includes a relay lens group 154A configured to receive light 108B from the dichroic mirror 116A. The relay lens group 154A is configured to focus light 108B at infinity. The third optical element 118C also includes another mirror 156 configured to guide light 108B from the relay lens group 154B to the image sensor 126. The relay lens group 154B is configured to converge and focus light 108B.

[0057] The optical instrument 100 also includes a plurality of optical filters 146B disposed within a filter selection module 133. The filter selection module 133 is operable (e.g., rotatable) to position a specific optical filter 146B (or without a filter) between the relay lens group 154A and the relay lens group 154B. Each optical filter 146B may have different wavelength-dependent transmission characteristics. Via the filter selection module 133, the optical instrument 100 is configured to insert one or more optical filters 146B between the relay lens group 154A and the relay lens group 154B, and is configured to remove one or more optical filters 146B from between the relay lens group 154A and the relay lens group 154B. Typically, at least some of the optical filters 146B are configured to reduce the transmission of stray light 108A entering the third optical element 118C.

[0058] A first optical element 118A (specifically a parabolic mirror 144 and a rectangular microlens array 149) defines a first optical axis along which light 108A propagates. A second optical element 118B defines a second optical axis along which light 108A and light 108B propagate. A third optical element 118C defines a third optical axis along which light 108B primarily propagates. Figure 9 As shown, the first optical axis, the second optical axis, and the third optical axis are parallel to each other.

[0059] The optical instrument 100 is configured to operate in wide-field fluorescence mode, confocal fluorescence mode, bright-field mode, reflection mode, and phase-contrast mode.

[0060] In wide-field fluorescence mode, biological sample 20 is illuminated by light 108A to induce fluorescence in the form of light 108B. Plate 120 is removed from region 121, so that the image captured by image sensor 126 is a wide-field image without spatial filtering.

[0061] In confocal fluorescence mode, biological sample 20 is illuminated by light 108A to induce fluorescence in the form of light 108B. Plate 120 is inserted into region 121 such that the image captured by image sensor 126 is a spatially filtered confocal image.

[0062] In bright-field mode, the biological sample 20 is illuminated by the transmitted light module 138. Light 108B represents light transmitted through or around the biological sample 20. Plate 120 is removed from region 121, such that the image captured by image sensor 126 is a wide-field image without spatial filtering. Alternatively, plate 120 is rotated within region 121, such that the captured image is a spatially filtered confocal image.

[0063] In dark mode, the transmission light module 138 (see...) Figure 8 A ring aperture is used to block almost any light from the transmission light module 138 through window 104 to the objective lens 112A and to the image sensor 126. If the biological sample 20 is located between the transmission light module 138 and the objective lens 112A, the biological sample 20 refracts or scatters the light, thereby redirecting some of the light to the objective lens 112A and to the image sensor 126.

[0064] In reflection mode, biological sample 20 is illuminated by light 108A to induce reflected light in the form of light 108B. Plate 120 is removed from region 121, such that the image captured by image sensor 126 is a wide-field image without spatial filtering. Alternatively, plate 120 is rotated within region 121, such that the captured image is a spatially filtered confocal image.

[0065] Phase-contrast mode is achieved at least in part through computation. Optical instruments 100 operating in phase-contrast mode typically involve the use of an annular aperture in a transmission light module 138 and include a phase plate as part of a third optical element 118C and / or an objective lens 112.

[0066] Figure 11 This is a cross-sectional schematic diagram of the transmissive light module 138. The transmissive light module 138 includes a light source 130B (e.g., one or more LEDs) and a condenser lens 142 located between the window 104 and the light source 130B. The transmissive light module 138 also includes a fine-particle diffuser 147 located between the window 104 and the condenser lens 142. The transmissive light module 138 also includes a filter 146A located between the window 104 and the condenser lens 142. The distance between the light source 130B and the condenser lens 142 can be adjusted by a worm gear 151 and a threaded lens adapter 153.

[0067] Figure 12 This is a transparent view of the transmissive light module 138. The worm shaft 155 is rotated (e.g., by a screwdriver) to drive the worm wheel 151 to rotate, which in turn causes the threaded lens adapter 153 to move the condenser lens 142 up and down.

[0068] Referring to the foregoing figures, a method of operating the optical instrument 100 is described below. For example, the optical instrument 100 moves an optical module 114 within a housing 102A relative to a stage 136 supporting biological samples 20A (e.g., live cells) and 20B (e.g., live cells), thereby aligning the optical module 114 with the biological sample 20A. While aligned with the biological sample 20A, the optical module 114 captures one or more first images of the biological sample 20A. Subsequently, the optical instrument 100 again moves the optical module 114 relative to the stage 136, thereby aligning the optical module 114 with the biological sample 20B. While aligned with the biological sample 20B, the optical module 114 captures one or more second images of the biological sample 20B.

[0069] Figure 3 The illustration shows a biological sample 20A contained in a first container and a biological sample 20B contained in an adjacent second container. Other examples may also be used. Biological samples 20A and 20B may contain some of the same biological material, while also having at least some different biological materials. In various scenarios, the division between them may correspond to different fields of view captured by image sensor 126.

[0070] In various examples, one or more first images and / or one or more second images may be captured in wide-field fluorescence mode, confocal fluorescence mode, bright-field mode, reflectance mode, or phase-contrast mode.

[0071] In wide-field mode, capturing one or more first images can remove the plate 120 from the optical path of the optical instrument 100 (i.e., from...). Figure 9 The process is performed when region 121 is removed. In wide-field mode, capturing one or more first images can be performed while illuminating the biological sample 20A with optical module 114.

[0072] In some examples, Figure 8 The transmitted light module 138 shown is used to illuminate the biological sample 20. Accordingly, the optical instrument 100 moves the transmitted light module 138 (e.g., synchronously with the optical module 114) so ​​that the biological sample 20A is positioned between the transmitted light module 138 and the optical module 114. In this case, one or more first images are captured while illuminating the biological sample 20A with the transmitted light module 138. Furthermore, the plate 120 can be rotated within region 121 so that the one or more first images become confocal images focused at different depths on the biological sample 20A.

[0073] In some examples, different excitation wavelengths can be used. Accordingly, the optical module 114 can illuminate the biological sample 20A with a first monochromatic light having a first wavelength, followed by illuminating the biological sample 20A with a second monochromatic light having a second wavelength. In this case, one or more first images are captured while the biological sample 20A is illuminated with the first monochromatic light. The optical module 114 also captures another image while the biological sample 20A is illuminated with the second monochromatic light.

[0074] In wide-field mode, one or more second images can be captured when plate 120 is removed from the optical path of optical instrument 100. In wide-field mode, one or more second images can be captured when biological sample 20B is illuminated with optical module 114.

[0075] In some examples, the transmitted light module 138 is used to illuminate the biological sample 20B. Accordingly, the optical instrument 100 moves the transmitted light module 138 so that the biological sample 20B is positioned between the transmitted light module 138 and the optical module 114. In this case, capturing one or more second images is accomplished while illuminating the biological sample 20B with the transmitted light module 138. Furthermore, the plate 120 can be rotated within region 121 so that one or more second images become confocal images focused at different depths on the biological sample 20B.

[0076] In some examples, different excitation wavelengths can be used. Accordingly, the optical module 114 can illuminate the biological sample 20B with a first monochromatic light having a first wavelength, followed by illuminating the biological sample 20B with a second monochromatic light having a second wavelength. In this case, one or more second images are captured while the biological sample 20B is illuminated with the first monochromatic light. The optical module 114 also captures another image while the biological sample 20B is illuminated with the second monochromatic light.

[0077] Figure 13 This is a block diagram of a method 300 for operating an optical instrument 100. (Example) Figure 13 As shown, method 300 includes one or more operations, functions, or actions illustrated in blocks 302, 304, 306, and 308. Although the blocks are shown sequentially, they may also be executed in parallel and / or in a different order than described herein. Furthermore, the blocks may be combined into fewer blocks, divided into more blocks, and / or deleted according to desired implementation methods.

[0078] At frame 302, method 300 includes optical instrumentation 100 that moves optical module 114 relative to stage 136 supporting biological samples 20A and 20B, thereby aligning optical module 114 with biological sample 20A. Optical module 114 includes plate 120 having an opening 131 for patterning in a rotary confocal microscope. Functions related to frame 302 have been referenced above. Figures 1 to 12 describe.

[0079] At box 304, method 300 includes capturing one or more first images of the biological sample 20A using an optical module. The functionality associated with box 304 has been referenced above. Figures 1 to 12 describe.

[0080] At box 306, method 300 includes optical instrument 100 moving optical module 114 relative to stage 136, thereby aligning optical module 114 with biological sample 20B. The functions associated with box 306 have been referenced above. Figures 1 to 12 describe.

[0081] At box 308, method 300 includes capturing one or more second images of the biological sample 20B using optical module 114. The functionality associated with box 308 has been referenced above. Figures 1 to 12 describe.

[0082] While various exemplary aspects and implementations have been disclosed herein, other aspects and implementations will be apparent to those skilled in the art. The exemplary aspects and implementations disclosed herein are for illustrative purposes only and are not intended to be limiting; the true scope and spirit are specified in the claims.

Claims

1. An optical instrument, comprising: Housing, the housing comprising: Window; and A port configured to receive a first light via an optical cable; An objective lens, configured to be aligned with the window such that first light emanating from the housing through the window enters the objective lens, and second light from the objective lens enters the housing through the window; and An optical module, located within the housing, comprises: Dichroic mirror; One or more first optical elements, the one or more first optical elements being configured to guide the first light from the port to the dichroic mirror; One or more second optical elements, the one or more second optical elements being configured to guide the first light from the dichroic mirror through the window into the objective lens, and the one or more second optical elements being configured to guide the second light from the window into the dichroic mirror; A plate having an opening for patterning in a rotary confocal microscope, wherein the plate is configured to be inserted into, or removed from, a region between the dichroic mirror and one or more second optical elements; Image sensors; and One or more third optical elements, the one or more third optical elements being configured to direct the second light from the dichroic mirror to the image sensor.

2. The optical instrument according to claim 1, wherein the housing is sealed.

3. The optical instrument according to any one of claims 1-2, wherein the housing is a first housing, and the optical instrument further includes a second housing, the second housing including a plurality of vents, wherein the second housing is configured to accommodate the first housing, the objective lens, and the optical module.

4. The optical instrument according to any one of claims 1-3, wherein the optical instrument is configured to be placed in an environmental chamber, the environmental chamber being configured to maintain a first temperature within the environmental chamber, the first temperature being different from a second temperature outside the environmental chamber.

5. The optical instrument according to any one of claims 1-4, wherein the optical instrument is configured to be placed in an environmental chamber, the environmental chamber being configured to maintain a first humidity, the first humidity being different from a second humidity outside the environmental chamber.

6. The optical instrument according to any one of claims 1-5, wherein the optical instrument is configured to be placed in an environmental chamber, the environmental chamber being configured to maintain a first carbon dioxide partial pressure within the environmental chamber, the first carbon dioxide partial pressure being different from a second carbon dioxide partial pressure outside the environmental chamber.

7. The optical instrument according to any one of claims 1-3, further comprising: Environmental chambers; as well as A light source located outside the environmental chamber, wherein the light source is configured to provide the first light to the housing via the optical cable.

8. The optical instrument according to any one of claims 1-7, wherein the region is within a confocal plane relative to the image sensor.

9. The optical instrument according to any one of claims 1-8, wherein the dichroic mirror is a first dichroic mirror, the first dichroic mirror and the second dichroic mirror are interchangeable, and the second dichroic mirror has wavelength-dependent transmission or reflection characteristics different from those of the first dichroic mirror.

10. The optical instrument according to any one of claims 1-9, wherein the optical instrument is configured to operate in at least one of wide-field fluorescence mode, confocal fluorescence mode, bright-field mode, dark-field mode, reflection mode or phase-contrast mode.

11. The optical instrument according to any one of claims 1-10, wherein the housing is formed of one or more materials including acetal homopolymer resin, anodized aluminum oxide, or stainless steel.

12. The optical instrument according to any one of claims 1-11, wherein the housing is formed of one or more materials, said material having one or more of the following properties: resistance to moisture expansion, non-toxicity to cell culture, antioxidant, corrosion-resistant, or thermal expansion-resistant.

13. The optical instrument according to any one of claims 1-12, wherein the housing has an IP64 protection rating as specified by the International Electrotechnical Commission.

14. The optical instrument according to any one of claims 1-13, further comprising one or more second objectives configured to be aligned with the window.

15. The optical instrument according to any one of claims 1-13, wherein the objective lens is a first objective lens, and the optical instrument further comprises: One or more second objectives; as well as A selection module is operable such that the first objective lens and the one or more second objectives lenses can be selected to align with the window.

16. The optical instrument according to any one of claims 1-15, further comprising: frame; as well as A stage, which is mounted to the frame and configured to hold one or more biological samples. The optical module and the housing are configured to move relative to the stage.

17. The optical instrument of claim 16, wherein the stage is configured to hold at least one container, the container being configured to contain a biological sample.

18. The optical instrument according to any one of claims 16-17, further comprising: A light transmission module, located outside the housing and configured to emit a light beam toward the window. The stage is located between the transmission light module and the objective lens.

19. The optical instrument according to claim 18, wherein the transmission light module comprises: light source; as well as A condenser lens is located between the window and the light source.

20. The optical instrument of claim 19 further includes a fine-particle diffuser located between the window and the condenser lens.

21. The optical instrument of claim 20 further includes a filter located between the window and the condenser lens.

22. The optical instrument according to any one of claims 19-21, wherein the distance between the light source and the condenser lens is adjustable.

23. The optical instrument according to any one of claims 1-22, wherein the opening is configured to spatially filter the second light.

24. The optical instrument according to any one of claims 1-23, wherein the image sensor comprises a complementary metal-oxide-semiconductor (CMOS) image sensor.

25. The optical instrument according to any one of claims 1-24, wherein the housing of the window and the port is a main housing, wherein the main housing is configured to maintain a humidity level inside the main housing that is lower than that outside the main housing.

26. The optical instrument according to any one of claims 1-25, wherein the optical cable comprises one or more optical fibers.

27. The optical instrument according to any one of claims 1-26, wherein the housing is filled with inert gas or dehumidified air.

28. The optical instrument according to any one of claims 1-27, wherein the first light is a laser beam.

29. The optical instrument according to any one of claims 1-28, wherein the first light has a first wavelength, the second light does not include the first wavelength, and wherein the dichroic mirror is configured to transmit the first light and reflect the second light.

30. The optical instrument according to any one of claims 1-29, wherein the one or more first optical elements comprise a parabolic mirror configured to guide the first light received from the port to the dichroic mirror.

31. The optical instrument of claim 30, wherein the one or more first optical elements further comprises a rectangular microlens array configured to shape the first light received from the parabolic mirror.

32. The optical instrument of claim 31, wherein the one or more first optical elements further comprises a prism reflector configured to reflect the first light received from the rectangular microlens array.

33. The optical instrument of claim 32, wherein the one or more first optical elements further comprises a positive lens configured to shape and guide the first light received from the prism mirror to the dichroic mirror.

34. The optical instrument of claim 33, wherein the dichroic mirror is configured to transmit the first light from the positive lens.

35. The optical instrument according to any one of claims 1-34, wherein the one or more second optical elements comprise a prism reflector configured to reflect the first light from the dichroic mirror toward the window and the second light toward the dichroic mirror.

36. The optical instrument of claim 35, wherein the one or more second optical elements further comprises a barrel lens configured to shape the first light from the prism mirror and transmit the first light through the window to the objective lens.

37. The optical instrument of claim 36, wherein the barrel lens and the objective lens are configured to focus the first light onto the sample.

38. The optical instrument according to any one of claims 36-37, wherein the barrel lens is configured to shape the second light from the objective lens and transmit the second light to the prism reflector.

39. The optical instrument according to any one of claims 1-38, wherein the one or more third optical elements comprise one or more first relay lenses, the one or more first relay lenses being configured to receive the second light from the dichroic mirror.

40. The optical instrument of claim 39, wherein the one or more first relay lenses are configured to focus the second light at infinity.

41. The optical instrument of claim 40, wherein the one or more third optical elements further comprises: One or more second relay lenses; as well as One or more optical filters, each having different wavelength-dependent transmission characteristics, wherein the optical instrument is configured to insert the one or more optical filters between the one or more first relay lenses and the one or more second relay lenses, and wherein the optical instrument is configured to remove the one or more optical filters from between the one or more first relay lenses and the one or more second relay lenses.

42. The optical instrument of claim 41, wherein at least one of the one or more optical filters is configured to reduce the transmission of the first light.

43. The optical instrument according to any one of claims 41-42, wherein the one or more third optical elements include another mirror configured to direct the second light from the one or more second relay lenses to the image sensor.

44. The optical instrument according to any one of claims 41-43, wherein the one or more second relay lenses are configured to converge and focus the second light.

45. The optical instrument according to any one of claims 1-44, wherein the one or more first optical elements define a first optical axis, the one or more second optical elements define a second optical axis, and the one or more third optical elements define a third optical axis, wherein the first optical axis, the second optical axis, and the third optical axis are parallel to each other.

46. ​​A method for operating an optical instrument, the method comprising: The optical module is moved relative to the stage supporting the first and second biological samples, thereby aligning the optical module with the first biological sample, wherein the optical module includes a plate having an opening patterned for rotary confocal microscopy. One or more first images of the first biological sample are captured using the optical module; The optical module is moved relative to the stage to align it with the second biological sample. as well as One or more second images of the second biological sample are captured using the optical module.

47. The method of claim 46, wherein capturing the one or more first images comprises capturing the one or more first images in a wide-field fluorescence mode, a confocal fluorescence mode, a bright-field mode, a dark-field mode, a reflection mode, or a phase-contrast mode.

48. The method of claim 47, wherein capturing the one or more first images in wide-field mode comprises: The one or more first images are captured while the plate is being removed from the optical path of the optical instrument.

49. The method of claim 48, wherein capturing the one or more first images in wide-field mode comprises: The first image is captured while the first biological sample is illuminated with the optical module.

50. The method according to any one of claims 46-49, further comprising: The transmission light module is moved so that the first biological sample is positioned between the transmission light module and the optical module. Capturing the one or more first images includes capturing the one or more first images while illuminating the first biological sample with the transmission light module.

51. The method of claim 47, wherein capturing the one or more first images in turntable confocal mode comprises: The first image is captured while the plate is located in the optical path of the optical instrument.

52. The method of claim 51, wherein capturing the one or more first images in turntable confocal mode comprises: The one or more first images are captured at different depths of focus within the first biological sample.

53. The method of claim 46, further comprising: The first biological sample is irradiated with a first monochromatic light having a first wavelength; And subsequently The first biological sample is irradiated with a second monochromatic light having a second wavelength. Capturing the one or more first images includes: capturing an image while illuminating the first biological sample with the first monochromatic light, and capturing another image while illuminating the first biological sample with the second monochromatic light.

54. The method of claim 46, wherein capturing the one or more second images comprises: One or more second images are captured in wide-field fluorescence mode, confocal fluorescence mode, bright-field mode, dark-field mode, reflection mode, or phase-contrast mode.

55. The method of claim 54, wherein capturing the one or more second images in wide-field mode comprises: The one or more second images are captured while the plate is being removed from the optical path of the optical instrument.

56. The method of claim 55, wherein capturing the one or more second images in wide-field mode comprises: The one or more second images are captured while the second biological sample is illuminated with the optical module.

57. The method according to any one of claims 46-56, further comprising: The transmission light module is moved so that the second biological sample is positioned between the transmission light module and the optical module. Capturing the one or more second images includes capturing the one or more second images while illuminating the second biological sample with the transmission light module.

58. The method of claim 54, wherein capturing the one or more second images in turntable confocal mode comprises: The one or more second images are captured while the plate is located in the optical path of the optical instrument.

59. The method of claim 58, wherein capturing the one or more second images in turntable confocal mode comprises: The one or more second images are captured at different depths of focus within the second biological sample.

60. The method of claim 46, further comprising: The second biological sample is irradiated with a first monochromatic light having a first wavelength; And subsequently The second biological sample is irradiated with a second monochromatic light having a second wavelength. Capturing the one or more second images includes: capturing an image while illuminating the second biological sample with the first monochromatic light, and capturing another image while illuminating the second biological sample with the second monochromatic light.

61. The method according to any one of claims 46-60, wherein the first biological sample or the second biological sample comprises live cells.

62. The method according to any one of claims 46-61, wherein the optical module is located within the housing, the optical module comprising: Dichroic mirror; One or more first optical elements, the one or more first optical elements being configured to guide first light from the port to the dichroic mirror; One or more second optical elements, the one or more second optical elements being configured to guide the first light from the dichroic mirror through a window into the objective lens, and the one or more second optical elements being configured to guide the second light from the window into the dichroic mirror; A plate having an opening for patterning in a rotary confocal microscope, wherein the plate is configured to be inserted into, or removed from, a region between the dichroic mirror and one or more second optical elements; Image sensor; as well as One or more third optical elements, the one or more third optical elements being configured to direct the second light from the dichroic mirror to the image sensor.

63. The method according to any one of claims 46-61, wherein the method is performed by an optical instrument according to any one of claims 1-45.

64. A non-transitory computer-readable medium storing instructions that, when executed by a computing device operatively coupled to an optical instrument according to any one of claims 1-45, cause the optical instrument to perform the method according to any one of claims 46-62.

Images