Structured illumination of a sample

Abstract

This application relates to structured illumination of a sample. A system includes a light source, a first grating and a second grating, and at least one reflecting component that, in a first position, forms a first optical path originating from the light source and extending to the first grating and then to a subsequent component in the system, and that, in a second position, forms a second optical path originating from the light source and extending to the second grating and then to the subsequent component.

Description

[0001] This application is a divisional application of the application filed on September 18, 2019, with application number 201910882910.X and invention title "Structural Lighting for Samples".

[0002] Cross-references to related applications

[0003] This application claims priority to U.S. Provisional Application 62 / 733,330, filed September 19, 2018, and Dutch Application Serial No. N2022286, filed December 21, 2018; the contents of which are incorporated herein by reference. Technical Field

[0004] This application relates to, but is not limited to, structural lighting for samples. Background Technology

[0005] Structural illumination microscopy (SIM) has been used to improve the resolution of images obtained from samples. SIM utilizes several images of a sample with different fringe patterns, exposing different locations on the sample to a range of illumination intensities. In some cases, this process can be repeated by rotating the pattern orientation around the optical axis at individual angles. The captured images can be combined into a single image with an extended spatial frequency bandwidth, which can then be converted back to real space to generate an image with a higher resolution than that captured by conventional microscopy. Existing SIM methods may have one or more characteristics that increase system complexity, size, manufacturing cost, and / or operating cost. Summary of the Invention

[0006] In a first aspect, a system includes: a light source; a first grating and a second grating; and a first reflective component that forms a first optical path originating from the light source and extending to the first grating and subsequently to a subsequent component in the system at a first position, and forms a second optical path originating from the light source and extending to the second grating and subsequently to a subsequent component at a second position.

[0007] The implementation may include any or all of the following features. A first reflective assembly includes a rotatable mirror assuming a first or second position. The rotatable mirror is bilateral and includes an elongated member, wherein an axis is coupled to the elongated member substantially at its center. The axis is offset from and substantially parallel to a plane defined by first and second optical paths. When the rotatable mirror assumes the first position, a first end of the elongated member blocks a first path originating from a light source and extending to a second grating, and reflects first light originating from the light source toward the first grating. When the rotatable mirror assumes the first position, a second end of the elongated member does not block a second path from the first grating to a subsequent assembly. When the rotatable mirror assumes the second position, a second end of the elongated member blocks a second path from the second grating, and reflects second light from the second grating toward a subsequent assembly. When the rotatable mirror assumes the second position, a first end of the elongated member does not block the first path originating from a light source and extending to the second grating. The first and second gratings are oriented such that their respective normals are substantially antiparallel to each other, and wherein the axis is substantially aligned with the normals. A rotatable mirror reciprocates between first and second positions. A first reflecting assembly includes a first translational mirror that undergoes a first translation to the first position. The first reflecting assembly also includes a second translational mirror that undergoes a second translation to the second position. The first and second translations are substantially perpendicular to a plane defined by first and second optical paths. The first translation is substantially parallel to the plane defined by the first and second optical paths. The first translational mirror undergoes a second translation to the second position, wherein the second translation is substantially parallel to the plane defined by the first and second optical paths. The first reflecting assembly includes a rotatable prism presenting either the first or second position. A first grating and a second grating are positioned adjacent to each other, wherein the rotatable prism in the first position reflects first light along the first optical path toward the first grating, and wherein the rotatable prism in the second position reflects second light along the second optical path toward the second grating. The first and second gratings face a subsequent component. The first and second gratings are in fixed positions relative to the light source. The subsequent component is a phase selector. The system also includes a phase selector located between the light source and the first reflecting assembly. The phase selector is positioned relative to the light source. The system also includes a second reflective component located in each of the first and second optical paths, preceding the first and second gratings. Each of the first and second optical paths has a first optical path portion originating from the light source and extending to the second reflective component, wherein each of the first and second optical paths has a second optical path portion originating from a subsequent component, and wherein the first and second optical path portions are substantially parallel to each other.

[0008] In a second aspect, a system includes: a light source; a first grating and a second grating; and at least one reflector having a first position that blocks a first path originating from the light source and extending to the second grating, and directs first light to the first grating without blocking a second path from the first grating to a subsequent component in the system, and the reflector having a second position that blocks a third path from the second grating, and directs second light from the second grating to a subsequent component without blocking the first path.

[0009] The implementation may include any or all of the following features: The respective grating orientations of the first and second gratings are substantially perpendicular to each other. The first and second gratings face each other. A subsequent component is a phase selector. The system also includes a phase selector located between the light source and at least one reflector.

[0010] In a third aspect, a method includes: positioning at least one reflective component to define a first optical path originating from a light source and extending to a first grating and subsequently to a subsequent component; directing first phase-selective light from the first optical path onto a sample; positioning at least one reflective component to define a second optical path originating from a light source and extending to a second grating and subsequently to a subsequent component; and directing second phase-selective light from the second optical path onto the sample.

[0011] The implementation may include any or all of the following features. Positioning at least one reflective component to define a first optical path includes blocking a first path originating from a light source and extending to a second grating, and guiding first light to the first grating without blocking a second path from the first grating to a subsequent component. Positioning at least one reflective component to define a second optical path includes blocking a third path from the second grating, and guiding second light from the second grating to a subsequent component without blocking the first path.

[0012] It should be understood that all combinations of the foregoing concepts and the additional concepts discussed in more detail below (assuming these concepts do not contradict each other) are contemplated as part of the inventive subject matter disclosed herein. In particular, all combinations of the claimed subject matter appearing at the end of this disclosure are contemplated as part of the inventive subject matter disclosed herein. Attached Figure Description

[0013] Figure 1 This is a schematic diagram of an example system that facilitates structural imaging microscopy (SIM) and in which a phase selector is placed after the reflective component.

[0014] Figure 2 It shows that it can be implemented as Figure 1 An example of a rotating inline grating system (RIGS) with rotatable mirrors, which is part of a system.

[0015] Figure 3A yes Figure 2 The top view of the system shows the rotatable reflector in the first position.

[0016] Figure 3B yes Figure 3A The diagram shows a perspective detail of the rotatable mirror in its first position.

[0017] Figure 4A yes Figure 2 The top view of the system shows the rotatable reflector in the second position.

[0018] Figure 4B yes Figure 4A The diagram shows a perspective detail of the rotatable mirror in its second position.

[0019] Figure 5 It can be implemented as Figure 1 A schematic diagram of a system with one or more reflectors, which is part of a system to facilitate SIM.

[0020] Figure 6 It can be implemented as Figure 1 A schematic diagram of an example system with a translational mirror, which is part of the system.

[0021] Figures 7A-7B This schematically illustrates what can be implemented as Figure 6 An example of a vertical translation mirror that is part of a system.

[0022] Figures 8A-8B This schematically illustrates what can be implemented as Figure 6 An example of a horizontally shifting reflector that is part of a system.

[0023] Figure 9 It can be implemented as Figure 1 A schematic diagram of a system with a rotatable prism, which is part of the system.

[0024] Figure 10 An example of a method that can be used to locate one or more reflective components to perform SIM is shown.

[0025] Figure 11 This is a schematic diagram of an example system that facilitates SIM and in which a phase selector is placed in front of a reflective component.

[0026] Figure 12 This is a schematic diagram of an example system that can be used for biological and / or chemical analysis; Figure 1 The system can be Figure 12 Part of the system.

[0027] Figure 13 An example of the system is shown. Detailed Implementation

[0028] This document describes examples of systems and techniques, including but not limited to, those that can provide structural illumination through facilitated structural illumination microscopy (SIM). Such systems / techniques may offer one or more advantages over existing methods, such as those described below.

[0029] Imaging (e.g., using SIM) can be performed to analyze samples of any of a variety of materials. In some implementations, SIM imaging or another type of imaging can be performed as part of a biological or chemical analysis (e.g., a process for sequencing genetic material). In one example, this process can be a DNA sequencing process, such as sequencing-while-synthesizing or next-generation sequencing (also known as high-throughput sequencing). In another example, this process can be used to achieve genotyping. As those skilled in the art will recognize, genotyping involves determining differences in an individual's genetic makeup (genotype) by examining an individual's DNA sequence using bioassays and comparing it to the sequence of another individual or a reference sequence. This process can include fluorescence imaging, in which a sample of genetic material is illuminated with light (e.g., a laser beam) to trigger a fluorescence response via one or more tags on the genetic material. Some nucleotides may have fluorescent tags associated with the nucleotides to fluoresce in response to exposure to an energy source. The wavelength of the fluorescence response can be used to determine the presence of the corresponding nucleotide. The fluorescence response can be detected during sequencing and used to establish a record of the nucleotides in the sample.

[0030] SIM imaging is based on spatial structured light. For example, the structure can consist of or include a pattern in the illumination light, which helps improve the resolution of the acquired image. In some implementations, the structure can include a fringe pattern. Fringe patterns can be generated by reflecting or transmitting diffraction by illuminating a beam of light onto a diffraction grating (referred to as a grating for simplicity). The structured light can be illuminating the sample according to corresponding fringes that may appear according to a certain period. For example, an image of the sample can be obtained at different phases of the fringes in the structured light (sometimes referred to as the corresponding pattern phase of the image). This allows various locations on the sample to be exposed to a variety of illumination intensities. The pattern of the structured light can be rotated relative to the sample, and the aforementioned image can be captured for each rotation angle.

[0031] Different types of gratings can be used in a variety of implementations. A grating can include one or more forms of periodic structures. In some implementations, a grating can be formed by removing or omitting physical material from the substrate. In other implementations, filters or other non-physical materials can be implemented to form the grating. For example, a substrate can be provided with a set of slits and / or grooves to form the grating. In some implementations, a grating can be formed by adding material to the substrate. For example, a periodically spaced structure can be formed on the substrate using the same or different materials.

[0032] For SIM systems, the ability to process samples rapidly is preferred to facilitate high throughput. Faster SIM imaging allows the analytical system to achieve greater throughput; that is, more chemical or biological samples can be imaged within the same time frame. For high throughput, the system may illuminate a relatively large sample area with high-contrast stripes and / or rapidly switch between stripe orientations. For such a system to achieve high throughput, imaging must therefore be highly repeatable and reliable. High optical power can help maintain relatively short exposure times. Therefore, good light efficiency and a powerful light source can contribute to achieving high optical power.

[0033] In some SIM systems that project high-contrast fringes, coherent light sources may be used. In such systems, single-mode lasers can be coherent sources, but they may be prohibitively expensive in terms of the type of analysis being performed and / or the amount of power required. Other types of light sources, such as light-emitting diodes (LEDs) or arc lamps, may not provide sufficient coherence for this application. Therefore, multimode lasers can be a viable candidate as coherent sources, but this is associated with characteristics such as having a multimode spectral pattern. To achieve the desired uniformity with a multimode laser, the multimode laser output can be mode-scrambled. However, using mode-scrambled multimode lasers can result in multiple sources selectively exciting multiple gratings, which can increase cost and optical system complexity unless optical switches are used. Furthermore, using mode-scrambled multimode lasers may also lead to reliance on relay lens systems to achieve zero-order blocking for the desired fringe frequency and modulation contrast.

[0034] This paper describes the implementation of a structured illumination system for analyzing samples. Some such systems include a single light source, at least two fixed gratings, and a mechanism for guiding the entire beam of light from the light source onto one or more of the gratings. A phase selector can be used to select the pattern phase. To name just a few examples, the mechanism can include any of a rotating double-sided mirror, a non-rotating mirror, or a rotatable prism mirror. For a rotating double-sided mirror, the mirror blades can be positioned in a first position to reflect light only onto a first grating, and can be positioned in a second position to reflect light only onto a second grating. A non-rotating mirror can be translated to either the first or second position. A rotatable prism mirror can selectively guide light from the light source onto either the first or second grating.

[0035] The examples described herein offer advantages over previous methods. In some implementations, a rotatable mirror can be used to switch between the excitations of individual gratings. Such a rotatable mirror can switch between two optical paths using a single light source. Furthermore, the dimensions of this rotatable mirror can be configured such that errors in rotational position and / or thermal effects do not significantly alter the optical path. Reducing the impact of such rotational changes and / or thermal effects on the optical path allows the SIM imaging system to operate faster because components that move the rotatable mirror (e.g., motors) can operate faster due to less reliance on fine-tuning of positioning compared to systems implementing movable gratings or other components. If more than two optical paths are to be implemented, multiple rotatable mirrors can be used for multiple optical paths. In some implementations, the grating can be fixed in place rather than rotated, translated, or otherwise moved as part of the operation. To cite just two examples, this provides angular accuracy and stability because the grating does not require fine-tuning of its position. In some implementations, a single light source can be used because the rotatable mirror can rotate into or out of the corresponding optical path to block or not block the corresponding optical path from the single light source. By implementing selectively positioned reflective components for the optical subsystem of the SIM system, mode blocking can be omitted for multimode lasers, and the entire path of light can be transmitted or blocked. Such a system can also eliminate the situation where the different orders of light emitted from the grating are separated from each other at a particular level of the system (e.g., requiring the different orders of light to be focused at a level where one or more orders are blocked while one or more orders are allowed to propagate). That is, the grating can emit undiffracted light, referred to as the 0th order light, and can also emit diffracted light propagating on the opposite side of the 0th order light, referred to as the + / -1 order light, respectively. In a system using mode blocking, both the 0th and + / -1 order lights can be focused at the blocking level, where it is assumed that the 0th order light is blocked while the + / -1 order is not blocked. In a system without mode blocking, some or all orders of light can be focused elsewhere in the system (e.g., at the objective lens), and such a system can have a shorter optical path length. Furthermore, this system with selectively positioned reflective components can eliminate the need for optical switches, thereby reducing the number of components and complexity of the optical system. Moreover, such a system can increase overall compactness; for example, by omitting relay lens systems that facilitate the recombination of multiple grating paths.

[0036] Figure 1 An example of a system 100 that can facilitate SIM imaging is illustrated schematically. System 100 can be used in conjunction with one or more other examples described herein. Some components in this and other examples are conceptually shown as blocks or other general components; such components can be implemented as one or more separate or integrated components to perform the indicated functions.

[0037] System 100 includes a light source 102. The light source 102 can be selected based on the coherence and / or power output that system 100 is to achieve. For example, a multimode laser can be used as the light source 102.

[0038] System 100 includes an optical structuring component 104 that receives light from light source 102. In some implementations, the optical structuring component 104 facilitates the illumination of the received light onto one or more gratings, thereby producing a pattern of light stripes. For example, one or more reflective components may be used to guide the light onto suitable gratings and / or further direct the light to the next stage in system 100. An example of the optical structuring component 104 is described below. A beam 106 extending between light source 102 and optical structuring component 104 schematically illustrates the propagation of light. The optical structuring component 104 can generate structured light and provide the structured light to subsequent components in system 100.

[0039] In some implementations, the subsequent component is a phase selector 108 in system 100. Phase selector 108 can receive light from optical structuring component 104. In some implementations, phase selector 108 is used to select the pattern phase in which the image will be captured. For example, as described in more detail herein, phase selector 108 can facilitate selection among multiple candidate pattern phases based on desired sample illumination or according to a required resolution metric.

[0040] System 100 includes a projection lens 110 that can receive light from a phase selector 108. This light may be referred to as phase-selective light to indicate that the light corresponds to a selection of the phase of a particular pattern, for example, by means of the phase selector 108. The projection lens 110 may include one or more optical elements, such as lenses that adjust the phase-selective light before it illuminates the next stage in system 100.

[0041] System 100 includes a mirror 112 that at least partially reflects light from projection lens 110 toward objective lens 114. In some implementations, mirror 112 provides selective transmission, for example, to reflect one or more portions of the illumination light arriving from projection lens 110 and to transmit at least a portion of the imaging light arriving from objective lens 114. For example, mirror 112 may be a dichroic mirror.

[0042] Objective lens 114 receives illumination light from mirror 112. Objective lens 114 may include one or more optical elements, such as lenses that adjust (e.g., reflected by mirror 112) the light from projection lens 110 before it illuminates the next stage in system 100.

[0043] Objective lens 114 directs light onto sample 116. In some implementations, sample 116 comprises one or more materials to be analyzed. For example, sample 116 may include genetic material to be irradiated for detecting a fluorescence response. Sample 116 may be held on a suitable substrate, including but not limited to a flow cell that allows selective flow of liquids or other fluids relative to the sample. For example, sample 116 may be subjected to a reagent containing one or more nucleotides before illumination, followed by image capture and analysis.

[0044] Sample 116 may be held by stage 118 in system 100. Stage 118 may provide one or more types of manipulation relative to sample 116. In some implementations, physical movement of sample 116 may be provided. For example, stage 118 may reposition sample 116 translationally and / or rotationally relative to at least one other component of system 100. In some implementations, thermal treatment of sample 116 may be provided. For example, stage 118 may heat and / or cool sample 116.

[0045] Stage 118 can facilitate phase selection. In some implementations, stage 118 can (e.g., using a piezoelectric actuator in stage 118) translate sample 116 a distance relative to a fixed light stripe to achieve phase selection. For example, phase selector 108 can then be bypassed in system 100 or eliminated from system 100.

[0046] In other words, light originating from light source 102 and modulated in the described components can be directed to sample 116 for illumination after propagating through objective lens 114. Any light emitted by sample 116 can pass through objective lens 114 in the opposite direction and be partially or completely transmitted through mirror 112. System 100 may include filter assembly 120 that receives light passing through mirror 112 and from objective lens 114. Filter assembly 120 may filter this light in one or more ways. For example, filter assembly 120 may allow certain wavelengths to pass through and / or block (or reflect) some other specific wavelengths. In some implementations, mirror 112 may incorporate filter assembly 120 as part of the mirror, for example by positioning filter assembly 120 on the rear surface of mirror 112.

[0047] Light passing through filter assembly 120 can enter camera system 122 in system 100. Camera system 122 may include one or more image sensors capable of detecting electromagnetic radiation of a type relevant to the analysis to be performed. In some implementations, camera system 122 is configured to capture images using fluorescence. For example, camera system 122 may include charge-coupled devices, complementary metal-oxide-semiconductor devices, or other image capture devices. Camera system 122 may produce output in digital and / or analog form. For example, data corresponding to images captured by camera system 122 may be stored by camera system 122 or may be sent to a separate component (e.g., a computer system or other device) for storage and / or analysis.

[0048] The operation of system 100 or other devices or machines will be illustrated below. In some implementations, the optical structuring assembly 104 includes one or more reflective components and at least one grating. For example, the reflective components may redirect light to or from the grating to produce light modulated to provide one or more forms of illumination for sample 116. In some implementations, the optical structuring assembly 104 may modulate light from light source 102 to perform SIM imaging. For example, such structured light may not need to be focused at a specific location within the optical structuring assembly 104; more precisely, the structured light (e.g., fringes of a diffraction pattern) may be focused at another stage of system 100, including but not limited to focusing on the back of objective lens 114.

[0049] Figure 2 An example of a system 200 with a rotatable reflector 202 is shown. System 200 can be used in conjunction with one or more other examples described herein. Individual components of system 200 can perform similar or identical functions to corresponding components described with reference to another example in this specification.

[0050] System 200 includes a light source 204. In some implementations, the light source 204 provides light, which in turn receives light through at least one optical fiber cable 206. For example, the light source 204 and the optical fiber cable 206 can be collectively referred to as an optical fiber transmitting module.

[0051] System 200 includes grating 208 and grating 210. In some implementations, grating 208 and / or 210 can serve as diffraction components with respect to light from light source 204. For example, grating 208 and / or 210 may include a substrate with a periodic structure, which is combined with a prism. Gratings 208 and 210 can be positioned relative to each other according to one or more arrangements. Here, gratings 208 and 210 face each other in system 200. Gratings 208 and 210 can be substantially identical to each other, or they can have one or more differences. The size, periodicity, or other spatial aspect of one grating of gratings 208 and 210 may differ from the size, periodicity, or other spatial aspect of the other grating. The grating orientation (i.e., the spatial orientation of the periodic structure) of one grating of gratings 208 and 210 may differ from the grating orientation of the other grating. In some implementations, the respective grating orientations of gratings 208 and 210 (which themselves face each other) can be substantially perpendicular to each other or at any other angle relative to each other. In some implementations, gratings 208 and 210 may be in offset positions relative to the rotatable mirror 202. In some implementations, gratings 208 and / or 210 may be in fixed positions relative to the light source 204.

[0052] System 200 may include one or more components (e.g., as...) Figure 1 The phase selector 108) facilitates the selection of the sample to be applied (e.g., applied to the phase selector 108). Figure 1 Phase selection of the light from sample 116 in the image. Here, system 200 includes a piezoelectric stripe shifter 212. In some implementations, piezoelectric stripe shifter 212 may receive light from gratings 208 and / or 210 and may perform some or all of the phase selection with respect to that light. For example, piezoelectric stripe shifter 212 may be used to control the pattern phase of structured light that should be used to capture a particular image. Piezoelectric stripe shifter 212 may include a piezoelectric actuator. For example, a piezoelectric piston system may be used to achieve phase selection. Other methods may be used. For example, a tilted optical plate may be used for phase selection. For example, here, system 200 is implemented on plate 214, and one or more regions of plate 214 may be tilted to perform phase selection. As another example, one or more of gratings 208 and 210 may be moved (e.g., translated) for phase selection, for example, by means of a piezoelectric actuator. The light emitted from piezoelectric stripe shifter 212 is sometimes referred to as phase-selected light to indicate that the light has been modulated according to a specific phase selection. In some implementations, gratings 208 and / or 210 may be in a fixed position relative to the light source 204.

[0053] The system includes a projection lens 216, which may include one or more optical components (e.g., lenses) to modulate light received from the piezoelectric stripe shifter 212. For example, the projection lens 216 may be positioned so that light enters the objective lens (e.g., Figure 1 The objective lens 114 controls the properties of light.

[0054] The rotatable reflector 202 can be used to redirect at least one beam to one or more of the gratings 208 or 210, and / or to redirect at least one beam arriving from one or more of the gratings 208 or 210. The rotatable reflector 202 may comprise one or more materials to sufficiently reflect electromagnetic waves that will be used to illuminate the sample. In some implementations, the light from the light source 204 comprises a laser beam of one or more wavelengths. For example, a metal-coated reflector and / or a dielectric reflector may be used. The rotatable reflector 202 may be bilateral. For example, the rotatable reflector 202 can be considered bilateral if it is capable of performing reflection at at least a portion on both sides (e.g., reflecting at a first end for a first beam path and at a second end opposite to the first end for a second beam path).

[0055] The rotatable reflector 202 may include an elongated member. The rotatable reflector 202 may have any of a variety of form factors or other shape characteristics. The rotatable reflector 202 may have a generally flat construction. The rotatable reflector 202 may have a generally square or other rectangular shape. The rotatable reflector 202 may have rounded corners. The rotatable reflector 202 may have a substantially constant thickness. The reflective surface of the rotatable reflector 202 may be substantially planar.

[0056] The rotatable mirror 202 may be supported by a shaft 218 of the system 200. The shaft 218 may allow the rotatable mirror 202 to rotate about the shaft 218 in one or both directions. The shaft 218 may be made of a material with sufficient rigidity to hold and manipulate the rotatable mirror 202, including but not limited to metals. The shaft 218 may be substantially coupled at the center of the rotatable mirror 202. For example, the rotatable mirror 202 may have an opening at the center, or a cutout extending from one side to the center, to facilitate coupling with the shaft 218. As another example, the shaft 218 may include separate shaft portions coupled to corresponding faces of the rotatable mirror 202 without requiring any openings in the rotatable mirror 202. The shaft 218 may have at least one suspension 220. Here, the suspension 220 is located at the ends of the shaft 218 on both sides of the rotatable mirror 202. The suspension 220 may include bearings or other features that facilitate low-friction operation.

[0057] The rotatable reflector 202 can be actuated to present one or more positions. Any type of motor or other actuator can be used to control the rotatable reflector 202. In some implementations, a stepper motor 222 is used. The stepper motor 222 can be coupled to shaft 218 and used to rotate shaft 218, thereby rotating the rotatable reflector 202 and presenting it to the desired position. In some implementations, the rotatable reflector 202 rotates toward a new position in the same direction (e.g., about the axis of rotation of shaft 218, always clockwise or always counterclockwise). In some implementations, the rotatable reflector 202 reciprocates between two or more positions (e.g., about the axis of rotation of shaft 218, alternately clockwise or counterclockwise).

[0058] Figures 3A-3B It shows the relationship with Figure 2 Examples related to System 200 in the document. Figure 3A The system 200 is shown in a top view, while Figure 3B The system 200 is shown in perspective. The rotatable reflector 202 is... Figures 3A-3B Each of them is in the same position

[0059] Here, light source 204 generates light 300 that propagates toward grating 210. A rotatable mirror 202 is positioned (e.g., oriented about a rotational axis 218) such that a first end 302 of the rotatable mirror 202 does not block light 300. Currently, the first end 302 may be positioned closer to the observer than the light 300 can propagate in the plane shown in the figures. That is, the reflective surface 202A of the rotatable mirror 202 facing light source 204 currently does not block light 300 because the first end 302 does not obstruct the path of light 300. Therefore, light 300 (through air, vacuum, or another fluid) propagates until it reaches grating 210.

[0060] Light 300 interacts with grating 210 in one or more ways. In some implementations, light 300 undergoes diffraction based on grating 210. Here, light 304 is structured light (e.g., having one or more patterned fringes) emitted from grating 210 based on the interaction between light 300 and grating 210. Light 304 initially propagates substantially in a direction generally toward projection lens 216. However, the position of rotatable mirror 202 causes a second end 306 of rotatable mirror 202 to block light 304. The second end 306 can be opposite the first end 302. In some implementations, the first end 302 and the second end 306 can be positioned at any angle relative to each other, such as any angle between 0 degrees and 180 degrees. Currently, the second end 306 may be positioned approximately as close to the observer as light 304. That is, the reflective surface 202B of rotatable mirror 202 facing grating 210 blocks light 304 because the second end 306 blocks the path of light 304. Therefore, from light 304, the rotatable mirror 202 directs light 308 to the piezoelectric stripe shifter 212.

[0061] Piezoelectric fringe shifter 212 performs phase selection on light 308. For example, piezoelectric fringe shifter 212 selects the pattern phase that the sample will undergo under the current illumination (e.g., for the purpose of capturing one or more specific images). Light 310 is emitted from piezoelectric fringe shifter 212 and propagates toward and enters projection lens 216. Light 310 corresponds to the specific phase selection performed using piezoelectric fringe shifter 212. Therefore, light 310 can be characterized as phase-selected light. Light 310 can then continue to propagate through the system (e.g., as in...). Figure 1 (The same as in system 100), for example, to irradiate sample 116.

[0062] Here, the phase-selective electromagnetic wave characteristic of light 310 corresponds to the fact that light 300 is diffracted by grating 210 and phase selection is performed by piezoelectric stripe shifter 212. Furthermore, the intervention of grating 210 here is the result of the positioning of rotatable mirror 202 such that its second end 306 blocks light 304 while its first end 302 does not block light 300.

[0063] Now assume that the rotatable mirror 202 is instead placed in a different position. Figures 4A-4B It shows the relationship with Figure 2 Another example related to System 200. Figure 4A The system 200 is shown in a top view, while Figure 4B The system 200 is shown in perspective. The rotatable reflector 202 is... Figures 4A-4B Each of them is in the same position

[0064] Here, light source 204 generates light 300 that initially propagates toward grating 210. A rotatable mirror 202 is positioned (e.g., oriented about a rotational axis 218) such that a first end 302 of the rotatable mirror 202 blocks light 300. Currently, the first end 302 may be positioned approximately as close to the observer as light 300. That is, the reflective surface 202A of the rotatable mirror 202 facing light source 204 blocks light 300 because the first end 302 obstructs its path. Therefore, light 312 (through air, vacuum, or another fluid) propagates until it reaches grating 208.

[0065] Light 312 interacts with grating 208 in one or more ways. In some implementations, light 312 undergoes diffraction based on grating 208. Here, light 314 is structured light (e.g., having one or more patterned fringes) emitted from grating 208 based on the interaction between light 312 and grating 208. Light 314 propagates substantially in a direction toward piezoelectric fringe shifter 212. The position of rotatable mirror 202 is such that the second end 306 of rotatable mirror 202 does not block light 314. Currently, the second end 306 may be positioned closer to the observer than light 314. That is, neither the reflecting surface 202B of rotatable mirror 202 nor the reflecting surface 202C facing grating 208 currently blocks light 314 because the second end 306 does not obstruct the path of light 314. Therefore, light 314 propagates until it reaches piezoelectric fringe shifter 212.

[0066] Piezoelectric fringe shifter 212 performs phase selection on light 314. For example, piezoelectric fringe shifter 212 selects the pattern phase that the sample will undergo under the current illumination (e.g., for the purpose of capturing one or more specific images). Light 316 is emitted from piezoelectric fringe shifter 212 and propagates toward and enters projection lens 216. Light 316 corresponds to the specific phase selection performed using piezoelectric fringe shifter 212. Therefore, light 316 can be characterized as phase-selected light. Light 316 can then continue to propagate through the system (e.g., as in...). Figure 1 (The same as in system 100), for example, to irradiate sample 116.

[0067] Here, the phase-selective electromagnetic wave characteristic of light 316 corresponds to the fact that light 300 is diffracted by grating 208 and phase selection is performed by piezoelectric stripe shifter 212. Furthermore, the intervention of grating 208 here results in the positioning of rotatable mirror 202 such that its first end 302 blocks light 300 while its second end 306 does not block light 314. Through various rotations, rotatable mirror 202 can be repeatedly presented in different positions (e.g., respectively). Figures 3A-3B and Figures 4A-4B(Position). For example, the rotatable reflector 202 can be located in... Figures 3A-3B Location and Figures 4A-4B The position moves back and forth. As another example, the rotatable reflector 202 can rotate in the same direction (e.g., clockwise or counterclockwise from the perspective of the stepper motor 222) to repeatedly present... Figures 3A-3B Location and Figures 4A-4B Location.

[0068] As mentioned above, gratings 208 and 210 can have different grating orientations relative to each other. For example, gratings 208 and 210 can have grating orientations that are substantially perpendicular to each other. Therefore, the light 304 emitted from grating 210 ( Figure 3A ) and light 314 emitted from grating 208 Figure 4A Lights can have different properties. For example, the fringe pattern in one of the lights, 304 and 314, can differ from the fringe pattern in the other. Illuminating a sample with light of different structures (e.g., Figure 1 Sample 116 in the sample can facilitate the use of system 200 for SIM imaging.

[0069] The example above illustrates a system comprising: a light source (e.g., light source 204); a first grating (e.g., grating 210) and a second grating (e.g., grating 208); a phase selector (e.g., piezoelectric stripe shifter 212); and at least one reflective component (e.g., rotatable mirror 202). (e.g., as...) Figures 3A-3B As shown, in the first position, the reflective component forms (e.g., by means of a first end 302 that does not block light 300) a first optical path from the light source to the first grating and subsequently (e.g., by means of a second end 306 that blocks light 304) to the phase selector. In (e.g., as...) Figures 4A-4B In the second position (as shown), the reflective component forms (e.g., by blocking light 300 at the first end 302) a second optical path from the light source to the second grating and subsequently (e.g., by not blocking light 314 at the second end 306) to the phase selector.

[0070] The example above also illustrates a system comprising: a light source (e.g., light source 204); a first grating (e.g., grating 208) and a second grating (e.g., grating 210); a phase selector (e.g., piezoelectric stripe shifter 212); and at least one reflector (e.g., rotatable reflector 202). Specifically, the reflector has (e.g., as...) Figures 4A-4B The first position (as shown) blocks the first path from the light source to the second grating (e.g., via the first end 302), while (e.g., via the second end 306 that does not block light 314) does not block the second path from the first grating to the phase selector. The mirror has (e.g., as...) Figures 3A-3BThe second position (as shown) blocks the third path from the second grating and directs the second light (e.g., light 308) to the phase selector, while (e.g., by the first end 302 that does not block light 300) does not block the first path.

[0071] The examples in this paper involve using reflective components and one or more gratings to provide structured light suitable for SIM imaging. In some implementations, mechanical motion may be significant (e.g., by rotating a mirror or another reflective component). However, reasonable mechanical and kinematic tolerances can be provided. For example, less precision or no precision may be required regarding the starting or stopping position of the reflective components (e.g., mirrors or prism mirrors); and stability and repeatability can be provided (e.g., using rotatable mirrors) by using precision bearings (e.g., in suspension 220), precision spindles (e.g., in shaft 218), and / or precise mirrors (e.g., using rotatable mirror 202 with low runout and / or good flatness). Stability and repeatability can be made independent of components that may wear (e.g., guide rails and / or end stops).

[0072] Figure 5 Another example of a system 500 that can be used as part of a SIM imaging system is schematically shown. System 500 can be used in conjunction with one or more other examples described herein. System 500 includes a light source 502, a mirror 504, gratings 506 and 508, a mirror 510, a phase selector 512, and a projection lens 514. Individual components of system 500 can perform similar or identical functions to corresponding components described with reference to another example in this specification. Here, gratings 506 and 508 face each other. In some implementations, gratings 506 and 508 may have different grating orientations, including but not limited to grating orientations that are substantially perpendicular to each other or at any other angle relative to each other. In some implementations, gratings 506 and 508 may be in offset positions relative to mirrors 504 and / or 510.

[0073] A Cartesian coordinate system with corresponding x, y, and z axes is shown. Here, the x and y axes extend in the plane shown, and the z axis extends perpendicular to the x and y axes in the direction toward the observer.

[0074] Path 516 is marked between light source 502 and grating 508. In this example and others, the path can indicate the route the beam can take if it is not blocked by some structure. Path 517 is marked between mirror 504 and grating 506. Path 518 is marked between grating 506 and phase selector 512. In this example, path 520 is marked on the side extending from grating 508 to projection lens 514. Paths 516, 517, 518, and 520 are shown here in dashed lines.

[0075] Here, light source 502 generates light 522 along at least a portion of path 516. If the position of mirror 504 is such that mirror 504 does not block path 516 and does not block light 522, then light 522 can propagate along path 516 and reach grating 508. That is, mirror 504 can then be considered to form the optical path of light 522 extending from light source 502 to grating 508. On the other hand, if the position of mirror 504 is such that mirror 504 blocks path 516 and blocks light 522, then mirror 504 can reflect light 522, and light 524 can propagate along path 517 toward grating 506. The redirected light 524 is indicated here by a dashed line. That is, mirror 504 can then be considered to form the optical path of light 522 and light 524 extending from light source 502 to grating 506. Therefore, the reflector 504 can selectively redirect the light 522 from the light source 502 to one of the selected gratings 506 or 508 between the two paths based on the position of the reflector 504.

[0076] Reflector 510 can selectively redirect light from a selected grating in grating 506 or 508 to phase selector 512 based on the position of reflector 510. If reflector 504 does not block path 516, allowing light 526 to be emitted from grating 508, and the position of reflector 510 such that it blocks path 520 and blocks light 526 emitted from grating 508, then reflector 510 can reflect light 528 toward phase selector 512. That is, reflectors 504 and 510 can then be considered to cooperate in forming the optical path of light 522, light 526, and light 528 extending from light source 502 to phase selector 512. On the other hand, if mirror 504 blocks path 516, causing light 524 to be redirected to grating 506, and the position of mirror 510 ensures that mirror 510 does not block path 518 and does not obstruct light 530, then light 530 can propagate along path 518 and reach phase selector 512. Light 530 is indicated here by a dashed line. That is, mirrors 504 and 510 can then be considered to cooperate in forming the optical paths of light 522, light 524, and light 530, which extend from light source 502 to phase selector 512.

[0077] Paths 516, 517, 518, and 520 can define one or more planes depending on the orientation of the components of system 500. Here, the optical paths including light 522, light 526, and light 528 extend substantially in the xy plane as shown (e.g., in the plane of the figures). Similarly, the optical paths including light 522, light 524, and light 530 also extend substantially in the xy plane. At least one aspect of system 500 can be substantially aligned with one or more such planes. In some implementations, mirrors 504 and 510 are rotatable mirrors (e.g., Figure 2 This is part of a rotatable mirror 202. For example, such a rotatable mirror can rotate at least partially about an axis 532 schematically shown here between mirrors 504 and 510. Axis 532 can be substantially parallel to the plane of one or more light paths. For example, axis 532 can be offset from the plane in a certain direction (e.g., toward the observer, similar to...). Figure 2 (Positioning of central axis 218). In some implementations, one or more of the light rays 522, 524, 526, 528 and / or 530 can propagate along a plane forming an angle relative to the xy plane (i.e., propagating in a direction toward or away from the observer), thereby forming a light path with components on the x, y and z axes.

[0078] Figure 6 An example of a system 600 with a translational mirror 602 is schematically shown. The translational mirror 602 is schematically shown here using a dashed outline. An example of the translational mirror 602 will be given below. System 600 can be used in conjunction with one or more other examples described herein. System 600 includes a light source 604, gratings 606 and 608, and a phase selector 610. Individual components of system 600 can perform similar or identical functions to corresponding components described with reference to another example in this specification.

[0079] The translational mirror 602 may include one or more mirrors that can undergo translation (in one or more directions) as part of the operation of the system 600. The translational mirror 602 can be translated to a first position, in which it forms an optical path 612 from the light source 604 to the grating 606 and subsequently to the phase selector 610. The translational mirror 602 can be translated to a second position, in which it forms an optical path 614 from the light source 604 to the grating 608 and subsequently to the phase selector 610. Therefore, the translational mirror 602 can selectively redirect light from the light source 604 to the phase selector 610 between the two optical paths 612 and 614.

[0080] Gratings 606 and 608 can be placed at any of a variety of positions relative to each other. The orientation of grating 606 can be characterized using its normal 616. For example, normal 616 can be a vector defined as perpendicular to the optically active surface of grating 606. The orientation of grating 608 can be characterized using its normal 618. For example, normal 618 can be a vector defined as perpendicular to the optically active surface of grating 608. In some implementations, normals 616 and 618 are substantially aligned with each other. For example, normals 616 and 618 can be substantially antiparallel to each other (e.g., oriented toward each other). In other implementations, normals 616 and 618 can form an angle between them.

[0081] Optical paths 612 and 614 may define one or more planes depending on the orientation of the components of system 600. Here, each of optical paths 612 and 614 extends substantially within the plane of the figures. In other implementations, optical paths 612 and / or 614 may have one or more portions extending out of or into the plane of the figures. At least one orientation of system 600 may be substantially aligned with one or more such planes of optical paths 612 or 614. In some implementations, the translational mirror 602 may undergo a translation substantially perpendicular to the planes of optical paths 612 and 614. In some implementations, the translational mirror 602 may undergo a translation substantially parallel to the planes of optical paths 612 and 614. Combinations of these methods may be used. In some implementations, the first side of the translational mirror 602 may have a first reflection angle (e.g., to form an optical path 612), and the second side of the translational mirror 602 may have a second reflection angle (e.g., to form an optical path 614), wherein the first reflection angle is different from the second reflection angle.

[0082] Figures 7A-7B It schematically shows the relationship with Figure 6 Examples related to system 600 are provided. The described examples can be used in conjunction with one or more other examples described herein. A translational mirror 602' is schematically shown here. The translational mirror 602' can be used as... Figure 6 The translational mirror 602 in the middle, or as a Figure 6 It is part of the translational mirror 602. That is, when a pair of mirrors 700 and 704 are in a position... Figure 7A The first translational position shown can form an optical path 614, while when in the position shown... Figure 7BIn the second translational position shown, the pair of mirrors 700 and 704 can form an optical path 612. The translational mirror 602' includes a mirror 700 coupled to a track 702. Here, mirror 700 has a rectangular shape, and side 700A of mirror 700 faces side 702A of track 702. Track 702 can facilitate vertical translation of mirror 700 along side 702A. For example, an actuator (not shown) can act on mirror 700 and reposition it in either direction along track 702. Similarly, the translational mirror 602' includes a mirror 704 having a rectangular shape and coupled to track 706 such that side 704A of mirror 704 faces side 706A of track 706. Therefore, track 706 can facilitate vertical translation of mirror 704 along side 706A. More than one track can be used to translate mirrors 700 and / or 704. Other types of actuation can be used. For example, mirrors 700 and / or 704 can be manipulated by actuators.

[0083] Figure 7A The configuration of a translational reflector 602' is shown, wherein reflector 700 is positioned toward end 708 of track 702, and reflector 704 is positioned toward end 710 of track 706. In some implementations, Figure 7A The position in the middle can correspond to the formation of one or more optical paths. For example, again refer to Figure 6 The position of the reflector 704 toward the end 710 can facilitate the blocking of the optical path 612 between the light source 604 and the grating 606. Due to this blocking, the reflector 704 can serve to redirect light from the light source 604 to the grating 608, and doing so can form the optical path 614. Furthermore, the reflector 700 currently positioned toward the end 708 may not block the optical path 614 between the grating 608 and the phase selector 610. Therefore, a translation of the reflector 602' to the configuration shown can form the optical path 614 in the system 600.

[0084] Figure 7B A configuration of a translational reflector 602' is shown, wherein reflector 700 is positioned toward end 712 of track 702, and reflector 704 is positioned toward end 714 of track 706. End 712 is substantially opposite to end 708, and end 714 is substantially opposite to end 710. In some implementations, Figure 7B The position in the middle can correspond to the formation of one or more optical paths. For example, again refer to Figure 6The position of mirror 704 towards end 714 may not block the light path 612 from light source 604, thus light may reach grating 606. Furthermore, mirror 700 currently positioned towards end 712 may block the light path 612 emanating from grating 606. Due to the obstruction, mirror 700 can serve to redirect light from grating 606 to phase selector 610, and doing so can form light path 612. Therefore, a translational mirror 602' to the configuration shown can form light path 612 in system 600. Figures 7A-7B Translation or self of the position shown in the figure Figures 7A-7B The translation of the position shown can occur in a direction that is substantially perpendicular to one or more planes of optical paths 612 and 614.

[0085] Figures 8A-8B It schematically shows the relationship with Figure 6 Another example related to system 600 is described herein. The described example may be used in conjunction with one or more other examples described herein. A "translatable reflector 602" is schematically shown here. "Translable reflector 602" can be used as... Figure 6 The translational mirror 602 in the middle, or as a Figure 6 This is part of a translational mirror 602. The translational mirror 602 includes a mirror 800 coupled to a track 802. Here, the mirror 800 has a rectangular shape, and the side 800A of the mirror 800 faces the side 802A of the track 802. The track 802 can facilitate translation of the mirror 800 along the side 802A. For example, an actuator (not shown) can act on the mirror 800 and reposition it in either direction along the track 802.

[0086] Figure 8A The configuration of a translational reflector 602” is shown, wherein the reflector 800 is positioned toward the end 804 of the track 802. In some implementations, Figure 8A The position in the middle can correspond to the formation of one or more optical paths. For example, again refer to Figure 6 The position of the reflector 800 toward end 804 facilitates the blocking of the optical path 612 between the light source 604 and the grating 606. Due to this blocking, the reflector 800 can redirect light from the light source 604 to the grating 608, thus forming the optical path 614. At end 806 of track 802, there is currently no positioned reflector. As a result, the translational reflector 602" may not block the optical path 614 between the grating 608 and the phase selector 610. Therefore, a translational mirror 602" to the configuration shown can form the optical path 614 in system 600.

[0087] Figure 8BThe configuration of a translational reflector 602” is shown, wherein the reflector 800 is positioned toward the end 806 of the track 802. In some implementations, Figure 8B The position in the middle can correspond to the formation of one or more optical paths. For example, again refer to Figure 6 The absence of a reflector at end 804 facilitates the propagation of light from source 604 to grating 606. Furthermore, the position of reflector 800 facing end 806 facilitates the blocking of light path 612 emanating from grating 606. Due to this blocking, reflector 800 can redirect light from grating 606 to phase selector 610, thus forming light path 612.

[0088] Figure 9 An example of a system 900 with a rotatable prism 902 is schematically shown. System 900 can be used in conjunction with one or more other examples described herein. System 900 also includes a light source 904, gratings 906 and 908, a phase selector 910, and a projection lens 912. Individual components of system 900 can perform similar or identical functions to corresponding components described with reference to another example in this specification.

[0089] The rotatable prism 902 can undergo rotation about one or more axes of rotation to present one or more positions. Here, the rotatable prism 902 can rotate about an axis perpendicular to the plane of the drawing, which is schematically represented by arrow 914. For simplicity, the rotatable prism 902 is shown here with a single orientation. However, the operation of the system 900 will be illustrated based on at least two different orientations of the rotatable prism 902. Here, gratings 906 and 908 face the phase selector 910. Other placements or orientations can be used.

[0090] Light source 904 provides light 916 that propagates toward rotatable prism 902. Light 916 interacts with rotatable prism 902 and undergoes reflection. Here, light 918 emitted from rotatable prism 902 is a result of this reflection when rotatable prism 902 is in a first position. Light 918 is guided to and interacts with grating 906. As a result of this interaction, light 920 is emitted from grating 906 and propagates toward and interacts with phase selector 910. As a result of this interaction, light 922 is emitted from phase selector 910 and propagates toward and interacts with projection lens 912. That is, when rotatable prism 902 is in the first position, it reflects light 918 along a first optical path from rotatable prism 902 toward grating 906.

[0091] Furthermore, the light 924 emitted from the rotatable prism 902 is a result of the reflection of light 916 when the rotatable prism 902 is in the second position. Light 924 is guided to and interacts with the grating 908. As a result of this interaction, light 926 is emitted from the grating 908 and propagates towards and interacts with the phase selector 910. As a result of this interaction, light 922 is emitted from the phase selector 910 and propagates towards and interacts with the projection lens 912. In other words, when the rotatable prism 902 is in the second position, it reflects light 924 along the second optical path from the rotatable prism 902 towards the grating 908.

[0092] Figure 10 An example of a method 1000 that can be used to perform a SIM is shown. Method 1000 can be performed in one or more systems illustrated herein. Method 1000 may include more or fewer operations than those shown. Unless otherwise stated, two or more operations of method 1000 may be performed in a different order. For illustrative purposes, some aspects of other examples described herein will be referenced.

[0093] In method 1000, method 1010 includes positioning the reflective component in a first position. The first position may facilitate the definition of a first optical path originating from a light source and extending to a first grating and subsequently to a subsequent component. For example, a rotatable reflector 202 may be placed... Figures 3A-3B The positions shown define the optical path from the light source 204 to the grating 210 and subsequently to the piezoelectric stripe shifter 212, the optical path including lights 300, 304, and 308. As another example, the rotatable mirror 202 can be placed... Figures 4A-4B The positions shown define the optical path from light source 204 to grating 208 and subsequently to piezoelectric stripe shifter 212, the optical path including lights 300, 312, and 314. As another example, Figure 5 The reflectors 504 and 510 can be positioned to define an optical path including beams 522, 526, and 528. As another example, Figure 5 Reflectors 504 and 510 can be positioned to define an optical path including beams 522, 524, and 530. As another example, a translational reflector 602' can be placed... Figure 7A The positions shown are for defining Figure 6 The optical path 614 in the middle. As another example, the translational mirror 602' can be placed in Figure 7B The positions shown are for defining Figure 6 The optical path 612 in the middle. As another example, the translational mirror 602” can be placed in Figure 8A The positions shown are for defining Figure 6The optical path 614 in the middle. As another example, the translational mirror 602” can be placed in Figure 8B The positions shown are for defining Figure 6 The optical path 612 in the example. As another example, Figure 9 The rotatable prism 902 can be positioned to define the optical path including beams 916, 918, 920, and 922. As another example, Figure 9 The rotatable prism 902 can be positioned to define the optical path including light beams 916, 924, 926, and 922. For example, subsequent components could be... Figure 1 The phase selector 108 in the example. As another example, subsequent components could be... Figure 11 The projection lens 110 in the middle.

[0094] In 1020, method 1000 includes directing a first phase-selective light from the first optical path onto the sample. For example, the phase-selective light can be drawn from piezoelectric stripe shifter 212 ( Figure 2 ) and / or from phase selector 108 ( Figure 1 ), 512 Figure 5 ), 610 Figure 6 ) or 910 ( Figure 9 One or more of the phase-selective light can be emitted. Figure 1 The sample 116 is then illuminated using phase-selective light (e.g., structured light). The sample can then be illuminated based on this phase-selective light (e.g., using...). Figure 1 The camera system 122 in the image is used for imaging, and for the sake of brevity, this operation is not explicitly discussed here.

[0095] In step 1030, method 1000 includes positioning the reflective component at a second position. This second position facilitates the definition of a second optical path originating from a light source and extending to a second grating and subsequently to a subsequent component. For example, a rotatable reflector 202 may be placed... Figures 3A-3B The positions shown define the optical path from the light source 204 to the grating 210 and subsequently to the piezoelectric stripe shifter 212, the optical path including lights 300, 304, and 308. As another example, the rotatable mirror 202 can be placed... Figures 4A-4B The positions shown define the optical path from light source 204 to grating 208 and subsequently to piezoelectric stripe shifter 212, the optical path including lights 300, 312, and 314. As another example, Figure 5 The reflectors 504 and 510 can be positioned to define an optical path including beams 522, 526, and 528. As another example, Figure 5Reflectors 504 and 510 can be positioned to define an optical path including beams 522, 524, and 530. As another example, a translational reflector 602' can be placed... Figure 7A The positions shown are for defining Figure 6 The optical path 614 in the middle. As another example, the translational mirror 602' can be placed in Figure 7B The positions shown are for defining Figure 6 The optical path 612 in the middle. As another example, the translational mirror 602” can be placed in Figure 8A The positions shown are for defining Figure 6 The optical path 614 in the middle. As another example, the translational mirror 602” can be placed in Figure 8B The positions shown are for defining Figure 6 The optical path 612 in the example. As another example, Figure 9 The rotatable prism 902 can be positioned to define the optical path including beams 916, 918, 920, and 922. As another example, Figure 9 The rotatable prism 902 can be placed at a position that defines the optical path including light 916, 924, 926 and 922.

[0096] In 1040, method 1000 includes directing a second phase-selective light from the second optical path onto the sample. For example, the phase-selective light can be drawn from piezoelectric stripe shifter 212 ( Figure 2 ) and / or from phase selector 108 ( Figure 1 ), 512 Figure 5 ), 610 Figure 6 ) or 910 ( Figure 9 One or more of the phase-selective light can be emitted. Figure 1 The sample 116 is then illuminated using phase-selective light (e.g., structured light). The sample can then be illuminated based on this second phase-selective light (e.g., using...). Figure 1 The camera system 122 in the image is used for imaging, and for the sake of brevity, this operation is not explicitly discussed here.

[0097] Figure 11 Another example of a system 1100 that can facilitate a SIM is illustrated schematically. System 1100 can be used in conjunction with one or more other examples described herein. Some components in this and other examples are conceptually shown as blocks or other general components; these components can be implemented as one or more separate or integrated components to perform the indicated functions. (Unspecified details regarding system 1100 are omitted.) Figure 1 The components corresponding to those components can play the same or similar roles in system 1100.

[0098] System 1100 includes a phase selector 108' located prior to optical structured component 104'. In some implementations, phase selector 108' may receive beam 106 from light source 102. Phase selector 108' may provide phase-selected light to optical structured component 104'. Optical structured component 104' may generate structured light and provide it to subsequent components in system 1100. In some implementations, the subsequent component is projection lens 110. Other methods may be used.

[0099] In some implementations, stage 118 can (e.g., using a piezoelectric actuator in stage 118) translate sample 116 a distance relative to a fixed light stripe to perform phase selection. For example, phase selector 108' can then be bypassed in or eliminated from system 1100.

[0100] Figure 12 This is a schematic diagram of an example system 1200 that can be used for biological and / or chemical analysis. In some implementations, the systems and / or techniques described herein (including, but not limited to, system 100) Figure 1 ) and / or method 1000 ( Figure 10 The carrier 1202 may be part of system 1200. System 1200 may be operable to obtain any information or data relating to at least one biological and / or chemical substance. In some implementations, the carrier 1202 supplies the material to be analyzed. For example, carrier 1202 may include a cartridge or any other component for holding the material. In some implementations, system 1200 has a container 1204 to receive carrier 1202 at least during analysis. Container 1204 may have an opening formed in the housing 1206 of system 1200. For example, some or all of the components of system 1200 may be located within housing 1206.

[0101] System 1200 may include an optical system 1208 for biological and / or chemical analysis of the material on carrier 1202. Optical system 1208 may perform one or more optical operations, including but not limited to illumination and / or imaging of the material. For example, optical system 1208 may include any or all of the systems described elsewhere herein. As another example, optical system 1208 may perform any or all of the operations described elsewhere herein.

[0102] System 1200 may include a thermal system 1210 for providing heat treatment in relation to biological and / or chemical analysis. In some implementations, thermal system 1210 thermally conditions at least a portion of the material to be analyzed and / or the support 1202.

[0103] System 1200 may include a fluid system 1212 for managing one or more fluids relevant to biological and / or chemical analysis. In some implementations, fluid may be supplied to the carrier 1202 or its material. For example, fluid may be added to and / or removed from the material of the carrier 1202.

[0104] System 1200 includes a user interface 1214 that facilitates inputs and / or outputs related to biological and / or chemical analysis. To name just a few examples, the user interface may be used to specify one or more parameters for the operation of system 1200 and / or for outputting results of biological and / or chemical analysis. For example, user interface 1214 may include one or more displays (e.g., a touchscreen), a keyboard, and / or a clicking device (e.g., a mouse or touchpad).

[0105] System 1200 may include a system controller 1216, which can control one or more aspects of system 1200 for performing biological and / or chemical analyses. System controller 1216 may control container 1204, optical system 1208, thermal system 1210, fluid system 1212, and / or user interface 1214. System controller 1216 may include at least one processor and at least one storage medium (e.g., memory) having executable instructions for the processor.

[0106] Figure 13 An example of a system 1300 with a rotatable reflector 1302 is shown. In some implementations, system 1300 may be characterized as a RIGS. System 1300 may be used in conjunction with one or more other examples described herein. Individual components of system 1300 may perform similar or identical functions to corresponding components described with reference to another example in this specification.

[0107] System 1300 includes a light source 1304. In some implementations, the light source 1304 provides light, which in turn receives light through at least one optical fiber cable 1306. For example, the light source 1304 and the optical fiber cable 1306 can be collectively referred to as an optical fiber transmitting module.

[0108] System 1300 includes grating 1308 and grating 1310. In some implementations, grating 1308 and / or 1310 can be used as diffraction components with respect to light from light source 1304. For example, grating 1308 and / or 1310 may include a substrate with a periodic structure, which is combined with a prism. Gratings 1308 and 1310 can be positioned relative to each other according to one or more arrangements. Here, gratings 1308 and 1310 face each other in system 1300. Gratings 1308 and 1310 can be substantially identical to each other, or can have one or more differences. The size, periodicity, or other spatial aspect of one grating of gratings 1308 and 1310 may differ from the size, periodicity, or other spatial aspect of the other grating. The grating orientation (i.e., the spatial orientation of the periodic structure) of one grating of gratings 1308 and 1310 may differ from the grating orientation of the other grating. In some implementations, the respective grating orientations (the gratings themselves facing each other) of gratings 1308 and 1310 can be substantially perpendicular to each other or at any other angle relative to each other. In some implementations, gratings 1308 and 1310 can be in an offset position relative to the rotatable mirror 1302. In some implementations, gratings 1308 and / or 1310 can be in a fixed position relative to the light source 1304.

[0109] System 1300 may include one or more components (e.g., as...) Figure 1 The phase selector 108) facilitates the selection of the sample to be applied (e.g., applied to the phase selector 108). Figure 1 Phase selection of light from sample 116 in the image. Here, system 1300 includes a piezoelectric stripe shifter 1312. In some implementations, piezoelectric stripe shifter 1312 may receive light from gratings 1308 and / or 1310 and may perform some or all of the phase selection with respect to that light. For example, piezoelectric stripe shifter 1312 may be used to control the pattern phase of a structured light that should be used to capture a particular image. Piezoelectric stripe shifter 1312 may include a piezoelectric actuator. For example, a piezoelectric piston system may be used to implement phase selection. Other methods may be used. For example, a tilted optical plate may be used for phase selection. For example, here, system 1300 is implemented on plate 1314, and one or more regions of plate 1314 may be tilted to perform phase selection. As another example, one or more of gratings 1308 and 1310 may be moved (e.g., translated) for phase selection, for example, by means of a piezoelectric actuator. The light emitted from the piezoelectric stripe shifter 1312 is sometimes referred to as phase-selective light to indicate that the light has been modulated according to a specific phase selection. In some implementations, gratings 1308 and / or 1310 may be in a fixed position relative to the light source 1304.

[0110] The system includes a projection lens 1316, which may include one or more optical components (e.g., lenses) to modulate light received from the piezoelectric stripe shifter 1312. For example, the projection lens 1316 may be positioned so that light enters the objective lens (e.g., Figure 1 The objective lens 114 controls the properties of light.

[0111] The rotatable mirror 1302 can be used to redirect at least one beam to one or more of the gratings 1308 or 1310, and / or to redirect at least one beam arriving from one or more of the gratings 1308 or 1310. The rotatable mirror 1302 may comprise one or more materials to sufficiently reflect the electromagnetic waves used to illuminate the sample. In some implementations, the light from the light source 1304 comprises a laser beam of one or more wavelengths. For example, a metal-coated mirror and / or a dielectric mirror may be used. The rotatable mirror 1302 may be bilateral. For example, the rotatable mirror 1302 can be considered bilateral if it is capable of performing reflection at at least a portion on both sides (e.g., reflecting at a first end for a first beam path and at a second end opposite to the first end for a second beam path).

[0112] The rotatable mirror 1302 may include an elongated member. The rotatable mirror 1302 may have any of a variety of form factors or other shape characteristics. The rotatable mirror 1302 may have a generally flat construction. The rotatable mirror 1302 may have a generally square or other rectangular shape. The rotatable mirror 1302 may have rounded corners. The rotatable mirror 1302 may have a substantially constant thickness. The reflective surface of the rotatable mirror 1302 may be substantially planar.

[0113] The rotatable mirror 1302 may be supported by a shaft 1318 of the system 1300. The shaft 1318 may allow the rotatable mirror 1302 to rotate about the shaft 1318 in one or both directions. The shaft 1318 may be made of a material with sufficient rigidity to hold and manipulate the rotatable mirror 1302, such material including, but not limited to, metals. The shaft 1318 may be substantially coupled at the center of the rotatable mirror 1302. For example, the rotatable mirror 1302 may have an opening at the center, or a cutout extending from one side to the center, to facilitate coupling with the shaft 1318. As another example, the shaft 1318 may include one or more separate shaft portions coupled to corresponding one or more faces of the rotatable mirror 1302 without requiring any opening in the rotatable mirror 1302. The shaft 1318 may have at least one suspension 1320. Here, the suspension 1320 is located at one end of the shaft 1318 on one side of the rotatable mirror 1302. The suspension 1320 may include bearings or other features that facilitate low-friction operation.

[0114] The rotatable reflector 1302 can be actuated to present one or more positions. Any type of motor or other actuator can be used to control the rotatable reflector 1302. In some implementations, a stepper motor 1322 is used. The stepper motor 1322 can be coupled to and used to rotate the shaft 1318, thereby rotating the rotatable reflector 1302 to present the desired position. In some implementations, the rotatable reflector 1302 rotates toward a new position in the same direction (e.g., about the axis of rotation of the shaft 1318, always clockwise or always counterclockwise). In some implementations, the rotatable reflector 1302 reciprocates between two or more positions (e.g., about the axis of rotation of the shaft 1318, alternately clockwise or counterclockwise).

[0115] The throughput and / or other performance characteristics of system 1300 may depend at least in part on the time taken when the rotatable reflector 1302 should change from one position to another. In some implementations, the type and / or manufacture of stepper motor 1322 may be selected at least in part based on the desired or anticipated performance of system 1300. For example, making stepper motor 1322 faster may allow for an increase in the switching speed of rotatable reflector 1302.

[0116] The cost of manufacturing and / or maintaining system 1300 may depend at least in part on the type of stepper motor 1322. In some implementations, stepper motor 1322 is a direct drive motor that directly drives shaft 1318 without requiring any gears or other intermediate components. For example, this implementation may reduce the number of parts and / or the cost of parts in system 1300.

[0117] Here, light source 1304 generates light 1324, which includes light 1324A propagating between light source 1304 and mirror 1326. Light 1324 is schematically shown in this figure to illustrate different propagation possibilities, and for clarity, the entire optical path is shown without being obscured by the structure of system 1300. Mirror 1326 can be used to reflect light 1324A to form light 1324B, which is directed to rotatable mirror 1302 and / or grating 1310. Mirror 1326 may comprise one or more materials to adequately reflect the electromagnetic waves used to illuminate the sample. In some implementations, the light from light source 1304 includes a laser beam of one or more wavelengths. For example, a metal-coated mirror and / or a dielectric mirror may be used.

[0118] The rotatable mirror 1302 is currently positioned (e.g., oriented about the axis of rotation of axis 1318) such that the first end 1328 of the rotatable mirror 1302 does not block light 1324B. Currently, the first end 1328 may be positioned closer to the observer than light 1324B might be propagating in the plane of the figure. That is, the reflective surface 1302A of the rotatable mirror 1302 facing the light source 1304 does not currently block light 1324B because the first end 1328 does not obstruct the path of light 1324B. Therefore, light 1324B (through air, vacuum, or another fluid) propagates until it reaches the grating 1310.

[0119] Light 1324B interacts with grating 1310 in one or more ways. In some implementations, light 1324B undergoes diffraction based on grating 1310. Here, light 1324C is structured light (e.g., having one or more patterned fringes) emitted from grating 1310 based on the interaction between light 1324B and grating 1310. Light 1324C initially propagates substantially in a direction generally toward the side of projection lens 1316. However, the position of rotatable mirror 1302 is such that a second end 1330 of rotatable mirror 1302 blocks light 1324C. The second end 1330 may be opposite the first end 1328. In some implementations, the first end 1328 and the second end 1330 may be positioned at any angle relative to each other, such as any angle between 0 degrees and 180 degrees. Currently, the second end 1330 may be positioned approximately as close to the observer as light 1324C. In other words, the reflective surface 1302B of the rotatable mirror 1302 facing the grating 1310 blocks the light 1324C because the second end 1330 blocks the path of the light 1324C. Therefore, according to the light 1324C, the rotatable mirror 1302 directs the light 1324D to the piezoelectric stripe shifter 1312.

[0120] Piezoelectric fringe shifter 1312 performs phase selection on light 1324D. For example, piezoelectric fringe shifter 1312 selects the pattern phase that the sample will undergo under the current illumination (e.g., for the purpose of one or more specific images). Light 1324E is emitted from piezoelectric fringe shifter 1312 and propagates toward and enters projection lens 1316. Light 1324E corresponds to the specific phase selection performed using piezoelectric fringe shifter 1312. Therefore, light 1324E can be characterized as phase-selected light. Light 1324E can then continue to propagate through the system (e.g., as in...). Figure 1 (The same as in system 100), for example, to irradiate sample 116.

[0121] Here, the phase-selective electromagnetic wave characteristic of light 1324E corresponds to the fact that light 1324B is diffracted by grating 1310 and phase selection is performed by piezoelectric stripe shifter 1312. Furthermore, the intervention of grating 1310 here is the result of the positioning of rotatable mirror 1302 such that its second end 1330 blocks light 1324C while its first end 1328 does not block light 1324B.

[0122] Now suppose the rotatable mirror 1302 is instead placed in a different position. Similar to the previous example, here the light source 1304 generates light 1324A that initially propagates toward mirror 1326. However, unlike the previous example, the rotatable mirror 1302 is positioned here (e.g., oriented about the axis of rotation 1318) such that the first end 1328 of the rotatable mirror 1302 blocks light 1324B. Currently, the first end 1328 may be positioned approximately as close to the observer as light 1324B. That is, the reflective surface 1302A of the rotatable mirror 1302 facing the light source 1304 blocks light 1324B because the first end 1328 blocks the path of light 1324B. Therefore, light 1324F (through air, vacuum, or another fluid) propagates until it reaches the grating 1308.

[0123] Light 1324F interacts with grating 1308 in one or more ways. In some implementations, light 1324F undergoes diffraction based on grating 1308. Here, light 1324G is structured light (e.g., having one or more patterned fringes) emitted from grating 1308 based on the interaction between light 1324F and grating 1308. Light 1324G propagates substantially in a direction toward piezoelectric fringe shifter 1312. The position of rotatable mirror 1302 is such that the second end 1330 of rotatable mirror 1302 does not block light 1324G. Currently, the second end 1330 may be positioned closer to the observer than light 1324G. That is, neither the reflective surface 1302B of rotatable mirror 1302 nor the reflective surface 1302C facing grating 1308 currently blocks light 1324G because the second end 1330 does not obstruct the path of light 1324G. Therefore, light 1324G propagates until it reaches the piezoelectric stripe shifter 1312.

[0124] The piezoelectric stripe shifter 1312 performs phase selection on the light 1324G. For example, the piezoelectric stripe shifter 1312 selects the pattern phase that the sample will undergo under the current illumination (e.g., for the purpose of capturing one or more specific images). Similar to the example described above, light 1324E is emitted from the piezoelectric stripe shifter 1312 and propagates toward and enters the projection lens 1316.

[0125] Here, the phase-selective electromagnetic wave characteristic of light 1324E corresponds to the fact that light 1324F is diffracted by grating 1308 and phase selection is performed by piezoelectric stripe shifter 1312. Furthermore, the intervention of grating 1308 here results in the positioning of rotatable mirror 1302 such that its first end 1328 blocks light 1324B while its second end 1330 does not block light 1324G. Through various rotations, rotatable mirror 1302 can repeatedly present different positions (e.g., the positions described in this example, respectively). For example, rotatable mirror 1302 can reciprocate between these positions. As another example, rotatable mirror 1302 can rotate in the same direction (e.g., clockwise or counterclockwise from the perspective of stepper motor 1322) to repeatedly present the positions.

[0126] As mentioned above, gratings 1308 and 1310 can have different grating orientations relative to each other. For example, gratings 1308 and 1310 can have grating orientations that are substantially perpendicular to each other. Therefore, light 1324C emitted from grating 1310 and light 1324G emitted from grating 1308 can have different characteristics. For example, the fringe pattern in one of the lights 1324C and 1324G can be different from the fringe pattern in the other light. Illuminating a sample (e.g., with light of different structured orientations) Figure 1Sample 116 in the sample can facilitate the use of system 1300 for SIM imaging.

[0127] One or more components of system 1300 may at least partially facilitate a design that reduces the space required to implement system 1300. For example, the design may include the geometry of one or more components of system 1300 selected to achieve space reduction. In some implementations, light 1324 travels along a path through system 1300 having a generally U-shaped geometry. For example, this design may facilitate the placement of light source 1304 and projection lens 1316 such that light 1324A and light 1324E propagate in substantially opposite directions to each other. In some implementations, mirror 1326 may facilitate the placement of gratings 1308 and 1310 within the space formed by light source 1304, stepper motor 1322, and projection lens 1316. In some implementations, mirror 1326 may facilitate the placement of gratings 1308 and 1310 substantially between light source 1304 and stepper motor 1322.

[0128] The example above illustrates a system comprising: a light source (e.g., light source 1304); a first grating (e.g., grating 1310) and a second grating (e.g., grating 1308); a phase selector (e.g., piezoelectric stripe shifter 1312); and at least one reflective component (e.g., rotatable mirror 1302). In a first position (e.g., as firstly illustrated), the reflective component forms a first optical path (e.g., by not blocking light 1324B at its first end 1328) from the light source to the first grating and subsequently (e.g., by blocking light 1324C at its second end 1330) to the phase selector. In a second position (e.g., as secondly illustrated), the reflective component forms a second optical path (e.g., by blocking light 1324B at its first end 1328) from the light source to the second grating and subsequently (e.g., by not blocking light 1324G at its second end 1330) to the phase selector.

[0129] The example above also illustrates a system comprising: a light source (e.g., light source 1304); a first grating (e.g., grating 1308) and a second grating (e.g., grating 1310); a phase selector (e.g., piezoelectric stripe shifter 1312); and at least one mirror (e.g., rotatable mirror 1302). Specifically, the mirror has (e.g., as illustrated below) a first position that (e.g., via a first end 1328) blocks a first path from the light source to the second grating, while (e.g., via a second end 1330 that does not block light 1324G) it does not block a second path from the first grating to the phase selector. The mirror has (e.g., as illustrated below) a second position that (e.g., via a second end 1330) blocks a third path from the second grating and directs second light (e.g., light 1324D) to the phase selector, while (e.g., via a first end 1328 that does not block light 1324B) it does not block the first path.

[0130] Reflector 1326 is an example of a reflective component that can be used in system 1300. The example above illustrates an implementation in which each of the first and second optical paths (e.g., illuminating gratings 1308 or 1310 respectively) has a first optical path portion (e.g., light 1324A) originating from a light source and extending to the second reflective component, wherein each of the first and second optical paths has a second optical path portion (e.g., light 1324E) originating from a subsequent component (e.g., piezoelectric stripe shifter 1312), and wherein the first and second optical path portions are substantially parallel to each other.

[0131] The examples in this paper involve using reflective components and one or more gratings to provide structured light suitable for SIM imaging. In some implementations, mechanical motion may be significant (e.g., by rotating a mirror or another reflective component). However, reasonable mechanical and kinematic tolerances can be provided. For example, less precision or no precision may be required regarding the starting or stopping position of the reflective components (e.g., mirrors or prism mirrors); and stability and repeatability can be provided (e.g., using rotatable mirrors) by using precision bearings (e.g., in suspension 1320), precision spindles (e.g., in shaft 1318), and / or precise mirrors (e.g., using rotatable mirror 1302 with low taper and / or good flatness). Stability and repeatability can be made independent of components that may wear (e.g., guide rails and / or end stops).

[0132] The terms “substantially” and “approximately” as used throughout this specification are used to describe and take into account small fluctuations, for example, due to variations in the process. For example, they may refer to less than or equal to ±5%, such as less than or equal to ±2%, such as less than or equal to ±1%, such as less than or equal to ±0.5%, such as less than or equal to ±0.2%, such as less than or equal to ±0.1%, such as less than or equal to ±0.05%. Furthermore, when used herein, indefinite articles such as “a” or “an” mean “at least one”.

[0133] It should be understood that all combinations of the foregoing concepts and the additional concepts discussed in more detail below (assuming these concepts do not contradict each other) are contemplated as part of the inventive subject matter disclosed herein. In particular, all combinations of the claimed subject matter appearing at the end of this disclosure are contemplated as part of the inventive subject matter disclosed herein.

[0134] Many implementations have been described. However, it will be understood that various changes may be made without departing from the spirit and scope of this specification.

[0135] Furthermore, the logical flow depicted in the accompanying drawings does not require the specific order or sequence shown to achieve the desired result. Additionally, other processes may be provided from the described flow, or processes may be eliminated, and other components may be added to or removed from the described system. Furthermore, other implementations are within the scope of the appended claims.

[0136] While certain features of the described implementations are illustrated herein, many modifications, substitutions, alterations, and equivalents will now occur to those skilled in the art. Therefore, it should be understood that the appended claims are intended to cover all such modifications and alterations falling within the scope of the implementations. It should be understood that they are given merely as examples and not as limitations, and various changes in form and detail are possible. Any part of the apparatus and / or method described herein can be combined in any combination (except mutually exclusive combinations). The implementations described herein may include various combinations and / or sub-combinations of the functions, components, and / or features of the different implementations described.

Claims

1. A system for structured illumination of a sample, comprising: light source; First grating; Second grating; A rotatable reflective assembly, wherein the rotatable reflective assembly includes a rotatable prism capable of rotating about a vertical axis between a first position and a second position; Wherein, when the rotatable prism is in the first position, the rotatable prism forms a first optical path including the light source and the first grating, and wherein, when the rotatable prism is in the second position, the rotatable prism forms a second optical path including the light source and the second grating; Wherein, the first grating and the second grating are in fixed positions relative to the light source; and The first grating is positioned substantially perpendicular to the second grating.

2. The system according to claim 1, wherein, The first grating includes a first substrate having a first periodic structure, and the second grating includes a second substrate having a second periodic structure.

3. The system according to claim 2, wherein, The first periodic structure has a first grating orientation and the second periodic structure has a second grating orientation, wherein the first grating orientation is substantially perpendicular to the second grating orientation.

4. The system according to claim 1, wherein, The first grating includes a first prism, and the second grating includes a second prism.

5. The system according to claim 1, wherein, The light source includes an optical fiber cable.

6. The system according to claim 1, wherein, The light from the light source includes laser beams of one or more wavelengths.

7. The system according to claim 1 further includes a projection lens.

8. The system according to claim 1 further includes a phase selector.

9. The system according to claim 8, wherein, The phase selector includes a piezoelectric actuator.

10. A system for structured illumination of a sample, comprising: light source; First grating; Second grating; A translational reflective component, the translational reflective component being capable of translating between a first position and a second position; Wherein, when the translational reflective component is in the first position, the translational reflective component forms a first optical path including the light source and the first grating, and wherein, when the translational reflective component is in the second position, the translational reflective component forms a second optical path including the light source and the second grating; and The first grating and the second grating are substantially perpendicular to each other.

11. The system according to claim 10, wherein, The first grating includes a first substrate having a first periodic structure, and the second grating includes a second substrate having a second periodic structure.

12. The system according to claim 11, wherein, The first periodic structure has a first grating orientation and the second periodic structure has a second grating orientation, wherein the first grating orientation is substantially perpendicular to the second grating orientation.

13. The system according to claim 10, wherein, The translational reflective assembly includes a translational reflector.

14. The system according to claim 13, wherein, The translational mirror is capable of horizontal translation relative to the first grating.

15. The system according to claim 13, wherein, The translational mirror can be translated vertically relative to the first grating.

16. The system of claim 10 further includes a phase selector.

17. The system according to claim 16, wherein, The phase selector includes a piezoelectric actuator.

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