Fluorescence microscope illumination method using pulsed xenon lamp, and fluorescence assembly

Abstract

A fluorescence microscope illumination method using a pulsed xenon lamp (304), and a fluorescence assembly. The fluorescence assembly comprises a wheel disc component (1), a camera component (2) and a light source component (3). The wheel disc component (1) comprises a wheel-disc mounting plate (101), wherein a plurality of three-hole fluorescence filter boxes (107) are provided on the wheel-disc mounting plate (101) by means of a central shaft (102) and a mounting shaft (104); a through hole (111) is provided in the wheel-disc mounting plate (101); and a wheel-disc driving mechanism drives the mounting shaft (104) to rotate at a fixed angle, such that the three-hole fluorescence filter boxes (107) take turns to stay directly below the through hole (111). The light source component (3) comprises the high-frequency pulsed xenon lamp (304), a lens (307) and a side rotating disc (313), wherein the lens (307) is located in the front of the high-frequency pulsed xenon lamp (304); the side rotating disc (313) is located in front of the lens (307); several light source filter holes (314) are provided in the side rotating disc (313); light source filters (315) are provided in the light source filter holes (314); and a second stepping electric motor (311) drives the side rotating disc (313) to rotate at a fixed angle, such that the light source filter holes (314) stay directly ahead of the lens (307) in turn. The fluorescence assembly can provide a microsecond-level high-luminance light source, and thus the photographing rate is greatly improved without lowering the image quality, the duration of preheating is short, and a high stability is achieved.

Description

A fluorescence microscope illumination method using a flash xenon lamp and a fluorescence component Technical Field

[0001] The present invention relates to the technical field of fluorescence microscopes, and in particular to a fluorescence microscope illumination method and a fluorescence component using a flash xenon lamp. Background Art

[0002] Fluorescence microscopy allows for a variety of imaging methods, including two-dimensional, three-dimensional, and time-series imaging of biological samples. Fluorescence microscopy allows for real-time observation of the specific structures and physiological functions of microscopic biological systems, such as living cells, animals, and bacteria. Consequently, fluorescence microscopy has found widespread application in biomedicine, medical diagnostics, and drug development.

[0003] (1) Application in biomedicine;

[0004] 1. Research on biological macromolecules: Fluorescent labeling can track and record changes in molecules in real time. Therefore, fluorescence microscopy is widely used in the study of protein, cell membrane, DNA and RNA structures. 2. Cell biology: Fluorescence microscopy can image physiological processes at the molecular level within cells, such as organelles, cell membranes, gene expression, and cell cycle, in real time. It can reveal the occurrence, development and termination of basic cellular physiological processes in both space and time, including cell migration, proliferation and apoptosis. 3. Neuroscience: Fluorescence microscopy can observe changes in microscopic structures such as nerve cells, nerve fibers and synapses in real time, including the morphology and distribution of synapses, neural discharges and neural imaging.

[0005] (2) Application in medical diagnosis;

[0006] 1. Biolabeling technology: fluorescent dyes are labeled onto biomarkers to be detected, such as viruses, cells, proteins, tumors, etc., which can be used for the detection and pathological diagnosis of various diseases; 2. Tissue sections: Fluorescence microscopes can efficiently image tissue sections to help medical scientists conduct more detailed research and diagnosis of diseases.

[0007] (3) Application in drug research and development;

[0008] 1. Drug analysis and screening: Fluorescence microscopy can help scientists quickly screen new drugs and determine their efficacy from the molecular to the cellular level. Fluorescent labeling can be used to analyze drug transport and metabolism, helping to explore the mechanism of drug action. 2. Toxicology research: Fluorescence microscopy can perform efficient three-dimensional imaging of biological molecules, cells, tissues, and mouse models to study the toxicity and side effects of chemicals.

[0009] Most current fluorescence microscopy technologies use high-pressure mercury lamps as the illumination source. Monochromatic light is obtained through a light source filter and then illuminated using epi-illumination. Fluorescence from the sample is then captured by a camera through a fluorescence filter, capturing the fluorescence information. However, the high-pressure mercury lamps used in current fluorescence microscopy are continuous light sources. Therefore, when the sample fluorescence is weak, the camera requires a longer exposure time (100ms or longer), which prevents the camera from increasing the image capture rate. This results in existing fluorescence microscopy technology being unable to achieve high-speed image scanning and rapid output of large sample volumes. Summary of the Invention

[0010] In response to the defects of the existing technology, the present invention provides a fluorescence microscope illumination method and fluorescence component using a flash xenon lamp, which can provide a microsecond-level high-brightness light source and greatly improve the shooting rate without reducing image quality.

[0011] In order to solve the above technical problems and achieve the above technical effects, the present invention is implemented through the following technical solutions:

[0012] A fluorescence microscope illumination method using a flash xenon lamp uses a high-frequency flash xenon lamp as a light source and performs high-frequency flash illumination on a sample on a glass slide through epi-illumination. The specific steps are as follows:

[0013] Step 1) using a high-frequency xenon flash lamp to emit a lateral flashing light beam;

[0014] Step 2) filtering the scintillation light beam using a light source filter;

[0015] Step 3) using a reflector to reflect the scintillation light beam filtered by the light source downward;

[0016] Step 4) the scintillation light beam reflected downward is irradiated onto a glass slide through an objective lens, wherein a portion of the scintillation light beam is reflected upward after contacting the sample;

[0017] Step 5) The upward reflected scintillation light beam passes through the objective lens and the reflector in sequence, and then passes through the fluorescence filtering of the fluorescence filter and enters the camera.

[0018] An electron microscope fluorescence assembly using a flash xenon lamp consists of a wheel component, a camera component and a light source component; wherein,

[0019] The wheel assembly includes a horizontally arranged wheel mounting plate, the lower surface of which is provided with a central axis pointing vertically downward, a mounting shaft driven by a wheel drive mechanism being rotatably sleeved on the central shaft, a plurality of three-hole fluorescence filter boxes being evenly arranged on the circumferential surface of the mounting shaft around its axis; a through hole is provided on the wheel mounting plate, and a detection position is located directly below the through hole. The wheel drive mechanism drives the mounting shaft to rotate at a fixed angle, thereby enabling all the three-hole fluorescence filter boxes to stay in the detection position in turn; the camera assembly is located directly above the through hole, and the camera assembly, the through hole, and the three-hole fluorescence filter box located at the detection position are vertically coaxial;

[0020] The light source component is located outside the detection position, and the light source component includes a high-frequency flashing xenon lamp, a lens and a side turntable driven by a second stepper motor. The lens is located directly in front of the high-frequency flashing xenon lamp, and the front of the lens is a light-transmitting position. The side turntable is located in front of the lens. A number of light source filter holes are evenly opened on the circumference of the disk surface of the side turntable, and each of the light source filter holes is covered with a light source filter; the second stepper motor drives the side turntable to rotate at a fixed angle, so that all the light source filter holes stay in the light-transmitting position in turn; the high-frequency flashing xenon lamp, the lens, the light source filter holes located at the light-transmitting position and the three-hole fluorescent filter box located at the detection position are transversely coaxial.

[0021] Furthermore, the wheel drive mechanism consists of a first stepper motor, a first spur gear, a second spur gear and an angle sensor. The first stepper motor is mounted vertically downward on the wheel mounting plate and is located on one side of the central shaft and the camera component. The first spur gear is fixedly mounted on the output shaft of the first stepper motor. The second spur gear is sleeved on the periphery of the central shaft and fixedly connected to the upper end face of the mounting shaft. The first spur gear and the second spur gear are at the same height and mesh with each other. The angle sensor includes a magnetic encoder, a central magnet, a magnet seat and a socket. The magnetic encoder is fixedly arranged on the axis of the lower end face of the central shaft. The magnet seat is fixedly connected to the lower end face of the mounting shaft. The central magnet is fixedly arranged inside the magnet seat and is located directly below the magnetic encoder. The cable of the magnetic encoder passes upward through the hollow inner cavity of the central shaft and is connected to the socket. The first stepper motor realizes the fixed angle rotation of the mounting shaft through the angular displacement measurement of the angle sensor, thereby realizing the alternating docking of all the three-hole fluorescent filter boxes with the lower end of the through hole.

[0022] Furthermore, the wheel disc component is provided with a first mechanical angle limiting mechanism, which includes a first flat spring piece, a first micro bearing seat, a first micro pin, a first roller, and a plurality of first roller positioning countersunk holes and a plurality of first roller rolling grooves opened on the plane circumference of the second spur gear; the inner end of the first flat spring piece is fixedly connected to the upper end face of the central shaft, the outer end of the first flat spring piece points in the direction of the detection position, the first micro bearing seat is fixedly arranged on the lower surface of the outer end of the first flat spring piece, and the first roller is installed on the first micro bearing seat through the first micro pin; the number of the first roller positioning countersunk holes and the first roller rolling grooves corresponds to the number of the three-hole fluorescent filter box, the opening position of the first roller positioning countersunk hole corresponds one-to-one to the position of the three-hole fluorescent filter box, the opening position of the first roller rolling groove is located between two adjacent first roller positioning countersunk holes, and the upper plane of the first roller positioning countersunk hole is higher than the upper plane of the first roller rolling groove, which is responsible for providing damping and card points for the first roller.

[0023] Furthermore, the three-hole fluorescent filter box includes a square box body, the top surface of the square box body is provided with an upper beam through hole for docking with the lower end of the through hole, the upper beam through hole is covered with a fluorescent filter, the bottom surface of the square box body is provided with a lower beam through hole for docking with the upper end of the objective lens, the upper beam through hole and the lower beam through hole are concentric above and below, a side beam through hole for docking with the inner end of the light source filter hole is provided on one side surface of the square box body, a reflector is obliquely arranged inside the square box body, and the light source component provides epi-illumination to the slide located below the objective lens through the reflector.

[0024] Furthermore, the circumferential surface of the mounting shaft is evenly arranged with a plurality of vertically arranged dovetail protrusions around its axis, and the number of the dovetail protrusions corresponds to the number of the three-hole fluorescent filter boxes. A dovetail groove that can be plugged into and matched with the dovetail protrusion is opened on one side of each of the three-hole fluorescent filter boxes, and the position of the dovetail groove is opposite to the position of the light beam side through-hole. A limiting piece is provided on the outer sleeve of the upper end of the mounting shaft, and the limiting piece is fixedly connected to the upper end faces of all the dovetail protrusions. The three-hole fluorescent filter box is connected to the dovetail groove through the dovetail groove. The cooperation of the tail protrusion realizes quick installation with the mounting shaft, and positioning is achieved by the limit plate; a C-shaped mounting groove is provided on the surface of each dovetail protrusion, and the notch of the C-shaped mounting groove extends to one side of the dovetail protrusion, and a ramp pressure block is provided in the C-shaped mounting groove, which can protrude or retract relative to the outer wall of the mounting shaft, and one edge of the ramp pressure block is located at the notch of the C-shaped mounting groove and is flush with the edge of the dovetail protrusion, and the dovetail groove is locked or unlocked with the dovetail protrusion by the retraction or protrusion of the ramp pressure block.

[0025] Furthermore, the camera component consists of a camera body, a reducing lens and a lens barrel, the lower end of the lens barrel is fixedly connected to the embedding groove opened around the upper end of the through hole, and the upper end of the lens barrel is connected to the lens of the camera body through the reducing lens.

[0026] Furthermore, the light source component includes a horizontally arranged light source mounting plate, a lamp holder seat is provided below the light source mounting plate through a lamp holder fixing seat, and the high-frequency flashing xenon lamp is fixedly installed in the lamp holder seat; a forward-extending C-shaped seat is provided at the bottom end of the lamp holder fixing seat, the lens is fixedly installed in the middle of the C-shaped seat through a lens fixing member, a side turntable connecting plate is provided at the front of the C-shaped seat, a connecting strip extending to one side is provided at the top of the side turntable connecting plate, the second stepper motor is fixedly installed on the lower surface of the connecting strip through a motor mounting plate, and the center of the outer side surface of the side turntable is fixedly connected to the output shaft of the second stepper motor through a connecting member.

[0027] Furthermore, a plurality of water-cooling coolers are fixedly attached to the outer surface of the lamp holder, and the outer cover of the lamp holder is provided with a protective cover for protecting the water-cooling coolers, and the rear end of the protective cover is provided with an opening for the coolant circulation pipeline to pass through.

[0028] Furthermore, a second mechanical angle limiting mechanism is provided on the light source component, and the second mechanical angle limiting mechanism includes a second flat spring piece, a second miniature bearing seat, a second miniature pin, a second roller, and a plurality of second roller positioning countersunk holes and a plurality of second roller rolling grooves opened on the rear side surface of the side turntable and arranged around its center; the inner end of the second flat spring piece is fixedly connected to the front side surface of the motor mounting plate through a flat spring piece fixing seat, the outer end of the second flat spring piece points in a direction away from the detection position, the second miniature bearing seat is fixedly arranged on the front side surface of the outer end of the second flat spring piece, and the second roller is mounted on the second miniature bearing seat through the second miniature pin; the number of the second roller positioning countersunk holes and the second roller rolling grooves corresponds to the number of the light source filter holes, the opening position of the second roller positioning countersunk hole corresponds one-to-one to the position of the light source filter hole, the opening position of the second roller rolling groove is located between two adjacent second roller positioning countersunk holes, and the upper plane of the second roller positioning countersunk hole is higher than the upper plane of the second roller rolling groove, which is responsible for providing damping and card points for the second roller.

[0029] Furthermore, the high-frequency flash xenon lamp can provide high-frequency flash light of 160-7500nm, with a frequency of up to 200Hz.

[0030] The beneficial effects of the present invention are:

[0031] 1. The present invention replaces the light source of the fluorescence microscope with a flash xenon lamp and adopts epi-illumination, thereby providing a microsecond-level high-brightness light source. Without reducing image quality, the shooting rate is greatly improved (200 frames / second), which is more than 20 times that of the existing technology. In addition, the warm-up time is short and the stability is good, which can realize high-speed fluorescence microscope scanning.

[0032] 2. The flash xenon lamp used in the present invention adopts a flashing working mode, so its service life is longer than that of a continuous light source and its corresponding cost is lower.

[0033] 3. The present invention adopts dual positioning of angle sensor and mechanical limit, so that the rotation angle of the fluorescent filter and the light source filter is more accurate and can be kept stable.

[0034] The fluorescent filter box of the present invention and the mounting shaft are assembled by adopting a dovetail slot plug-in method and fixed with an inclined pressure block, which greatly facilitates the disassembly and assembly of the fluorescent filter box and can also ensure the reliability of the installation.

[0035] The above description is only an overview of the technical solution of the present invention. In order to more clearly understand the technical means of the present invention and to implement it according to the contents of the description, the following preferred embodiments of the present invention are described in detail with reference to the accompanying drawings. The specific implementation methods of the present invention are given in detail by the following embodiments and the accompanying drawings. BRIEF DESCRIPTION OF THE DRAWINGS

[0036] The drawings described herein are used to provide a further understanding of the present invention and constitute a part of this application. The exemplary embodiments of the present invention and their descriptions are used to explain the present invention and do not constitute an improper limitation of the present invention. In the drawings:

[0037] FIG1 is a front perspective view of the present invention;

[0038] FIG2 is a perspective view of the back side of the present invention;

[0039] FIG3 is a bottom perspective view of the present invention;

[0040] FIG4 is a perspective view of a wheel disc component of the present invention;

[0041] FIG5 is a perspective view of the wheel disc component of the present invention after the wheel disc mounting plate is removed;

[0042] FIG6 is a schematic diagram of the assembly relationship between the three-hole fluorescent filter box and the mounting shaft in the wheel assembly of the present invention;

[0043] FIG7 is an exploded view of the central axis, mounting axis and angle sensor in the wheel disc assembly of the present invention;

[0044] FIG8 is a front perspective view of a three-hole fluorescence filter box in a wheel assembly of the present invention;

[0045] FIG9 is a front perspective view of a three-hole fluorescent filter box in a wheel assembly of the present invention;

[0046] FIG10 is a schematic diagram of the assembly relationship between the inclined pressure block and the mounting shaft in the wheel assembly of the present invention;

[0047] FIG11 is a structural diagram of a first mechanical angle limiting mechanism in a wheel disc assembly of the present invention;

[0048] FIG12 is a perspective view of a camera assembly according to the present invention;

[0049] FIG13 is a perspective view of the light source component of the present invention from the rear side;

[0050] FIG14 is a front side perspective view of a light source component of the present invention;

[0051] FIG15 is a schematic diagram showing the installation position of a water-cooling cooler in a light source component of the present invention;

[0052] FIG16 is a structural diagram of a second mechanical angle limiting mechanism in a light source component of the present invention;

[0053] FIG17 is a diagram showing the working principle of the present invention. DETAILED DESCRIPTION

[0054] The following will be described in detail with reference to the accompanying drawings to better understand the purpose, features and advantages of the invention. It should be understood that the embodiments shown in the accompanying drawings are not intended to limit the scope of the invention, but are only intended to illustrate the essential spirit of the technical solution of the invention.

[0055] In the following description, for the purpose of illustrating the various disclosed embodiments, certain specific details are set forth in order to provide a thorough understanding of the various disclosed embodiments. However, those skilled in the relevant art will recognize that the embodiments may be practiced without one or more of these specific details. In other cases, well-known devices, structures, and techniques associated with this application may not be shown or described in detail to avoid unnecessarily obscuring the description of the embodiments.

[0056] Unless the context requires otherwise, throughout the specification and claims, the word "comprise" and variations such as "include" and "have" should be construed in an open, inclusive sense, that is, should be interpreted to mean "including, but not limited to."

[0057] Reference throughout this specification to "one embodiment" or "an embodiment" means that a particular feature, structure, or characteristic described in connection with the embodiment is included in at least one embodiment. Thus, the appearances of "in one embodiment" or "in an embodiment" in various places throughout this specification are not necessarily all referring to the same embodiment. Furthermore, the particular features, structures, or characteristics may be combined in any manner in one or more embodiments.

[0058] As used in this specification and the appended claims, the singular forms "a," "an," and "the" include plural referents unless the context clearly dictates otherwise. It should be noted that the term "or" is generally employed in its sense including "and / or" unless the context clearly dictates otherwise.

[0059] In addition, the technical features involved in the different embodiments of the present invention described below can be combined with each other as long as they do not conflict with each other.

[0060] As shown in FIG17 , a fluorescence microscope illumination method using a flash xenon lamp is shown. A high-frequency flash xenon lamp is used as a light source to illuminate a sample on a glass slide using epi-illumination. The specific steps are as follows:

[0061] Step 1) using a high-frequency xenon flash lamp to emit a lateral flashing light beam;

[0062] Step 2) filtering the scintillation light beam using a light source filter;

[0063] Step 3) using a reflector to reflect the scintillation light beam filtered by the light source downward;

[0064] Step 4) the scintillation light beam reflected downward is irradiated onto a glass slide through an objective lens, wherein a portion of the scintillation light beam is reflected upward after contacting the sample;

[0065] Step 5) The upward reflected scintillation light beam passes through the objective lens and the reflector in sequence, and then passes through the fluorescence filtering of the fluorescence filter and enters the camera.

[0066] 1-3 , an electron microscope fluorescence assembly using a flash xenon lamp is mainly composed of a wheel component 1 , a camera component 2 and a light source component 3 .

[0067] As shown in Figures 4-7 , a preferred embodiment of the present invention comprises a horizontally disposed wheel mounting plate 101. A vertically downwardly extending central shaft 102 is disposed on the lower surface of the wheel mounting plate 101. A mounting shaft 104, driven by the wheel drive mechanism, is rotatably mounted on the central shaft 102 via two bearings 103. A locking nut 105 is disposed at the bottom of the central shaft 102. A bearing cover 106 is mounted around the outer periphery of the locking nut 105 and fixedly connected to the lower end surface of the mounting shaft 104. Multiple three-hole fluorescence filter boxes 107, for example, six, may be disposed on the circumferential surface of the mounting shaft 104, evenly spaced around its axis. A through hole 111 is provided on the wheel mounting plate 101 , and a detection position is located directly below the through hole 111 . The wheel driving mechanism drives the mounting shaft 104 to rotate at a fixed angle, thereby enabling all the three-hole fluorescence filter boxes 107 to stay in the detection position in turn.

[0068] As a preferred embodiment of the present invention, referring to Figures 8-9, the three-hole fluorescence filter box 107 includes a square box body 1071. The top surface of the square box body 1071 is provided with an upper beam through hole 1072 for docking with the lower end of the through hole 111, and the upper beam through hole 1072 is covered with a fluorescence filter 1073. The bottom surface of the square box body 1071 is provided with a lower beam through hole 1074 for docking with the upper end of the objective lens 4. The upper beam through hole 1072 and the lower beam through hole 1074 are concentric with each other. A side beam through hole 1075 is provided on one side surface of the square box body 1071 for docking with the inner end of the light source filter hole 314. A reflector 1076 is obliquely arranged inside the square box body 1071. The light source component 3 provides epi-illumination to the slide located below the objective lens 4 through the reflector 1076.

[0069] As a preferred embodiment of the present invention, referring to Figures 4-5, the wheel drive mechanism consists of a first stepper motor 108, a first spur gear 109, a second spur gear 110 and an angle sensor. The first stepper motor 108 is mounted vertically downward on the wheel mounting plate 101, and both are located on one side of the central shaft 102 and the camera component 2. The first spur gear 109 is fixedly mounted on the output shaft of the first stepper motor 108. The second spur gear 110 is sleeved on the periphery of the central shaft 102 and fixedly connected to the upper end face of the mounting shaft 104. The first spur gear 109 and the second spur gear 110 are at the same height and mesh with each other.

[0070] As a preferred embodiment of the present invention, referring to FIG7 , the angle sensor includes a magnetic encoder 112, a central magnet 113, a magnet seat 114 and a socket 115. The magnetic encoder 112 is fixedly arranged on the axis of the lower end surface of the central shaft 102. The magnet seat 114 is fixedly connected to the lower end surface of the mounting shaft 104. The central magnet 113 is fixedly arranged inside the magnet seat 114 and is located directly below the magnetic encoder 112. The cable of the magnetic encoder 112 passes upward through the hollow inner cavity of the central shaft 102 and is connected to the socket 115. The first stepper motor 108 measures the angular displacement of the angle sensor, thereby realizing a fixed-angle rotation of the mounting shaft 104, and further realizing the three-hole fluorescent filter box 107 staying in the detection position in turn.

[0071] Taking the arrangement of six three-hole fluorescent filter boxes 107 as an example, the angular displacement of the first stepper motor 108 during one rotation is set to 60°. Then, when the angle sensor measures that the angular displacement of the mounting shaft 104 reaches 60°, it is recorded as one rotation of the first stepper motor 108, thereby realizing the fixed angle rotation of the mounting shaft 104 in units of 60°, and further realizing the six three-hole fluorescent filter boxes 107 staying in the detection position in turn.

[0072] As shown in FIG11 , in addition to the angle sensor with a magnetic encoder as its core, the present invention also employs a first mechanical angle limiting mechanism. The first mechanical angle limiting mechanism comprises a first flat spring piece 116, a first micro-bearing seat 117, a first micro-pin 118, a first roller 119, and a plurality of first roller positioning countersunk holes 120 and a plurality of first roller rolling grooves 121 formed on the upper plane circumference of the second spur gear 110. The inner end of the first flat spring piece 116 is fixedly connected to the upper end surface of the central shaft 102, and the outer end of the first flat spring piece 116 points in the direction of the detection position. The first micro-bearing seat 117 is fixedly mounted on the lower surface of the outer end of the first flat spring piece 116, and the first roller 119 is mounted on the first micro-bearing seat 117 via the first micro-pin 118. The number of the first roller positioning countersunk holes 120 and the first roller rolling grooves 121 corresponds to the number of the three-hole fluorescence filter box 107, for example, six. The opening position of the positioning countersunk hole 120 corresponds one-to-one to the position of the three-hole fluorescent filter box 107, and the opening position of the first roller rolling groove 121 is located between two adjacent first roller positioning countersunk holes 120. The upper plane of the first roller positioning countersunk hole 120 is higher than the upper plane of the first roller rolling groove 121. The recessed first roller rolling groove 121 facilitates the rolling of the first roller 119, and the protruding first roller positioning countersunk hole 120 is responsible for providing damping for the first roller 119 and providing a card point for the first roller 119.

[0073] As a preferred embodiment of the present invention, referring to FIG. 6-10 , each of the three-hole fluorescence filter boxes 107 can be detachably mounted on the circumferential surface of the mounting shaft 104 . The circumferential surface of the mounting shaft 104 is evenly arranged with a number of vertically arranged dovetail protrusions 122 around its axis, and the number of the dovetail protrusions 122 corresponds to the number of the three-hole fluorescent filter boxes 107. A dovetail groove 123 that can be plugged into and cooperate with the dovetail protrusion 122 is opened on one side of each of the three-hole fluorescent filter boxes 107. The position of the dovetail groove 123 is opposite to the position of the beam side through-hole 1075. A limiting plate 124 is provided on the outer periphery of the upper end of the mounting shaft 104, and the limiting plate 124 is fixedly connected to the upper end surfaces of all the dovetail protrusions 122. The three-hole fluorescent filter box 107 is quickly installed on the mounting shaft 104 through the cooperation between the dovetail groove 123 and the dovetail protrusion 122, and positioning is achieved through the limiting plate 124. A C-shaped mounting groove 125 is provided on the surface of each dovetail protrusion 122, and the notch of the C-shaped mounting groove 125 extends to a side edge of the dovetail protrusion 122. A ramp pressure block 126 is provided in the C-shaped mounting groove 125, which can protrude or retract relative to the outer wall of the mounting shaft 104. An edge of the ramp pressure block 126 is located at the notch of the C-shaped mounting groove 125 and is flush with the edge of the dovetail protrusion 122. The dovetail groove 123 is locked or unlocked with the dovetail protrusion 122 by the retraction or protrusion of the ramp pressure block 126.

[0074] As shown in FIG12 , a preferred embodiment of the present invention, the camera assembly 2 is located directly above the through hole 111. The camera assembly 2 comprises a camera body 201, a zoom lens 202, and a lens barrel 203. The lower end of the lens barrel 203 is fixedly connected to a bezel formed around the upper end of the through hole 111, and the upper end of the lens barrel 203 is connected to the lens of the camera body 201 via the zoom lens 202.

[0075] As a preferred embodiment of the present invention, as shown in Figures 13-16, the light source component 3 is located outside the detection position. The light source component 3 includes a horizontally arranged light source mounting plate 301. Below the light source mounting plate 301, a lamp holder 303 is provided via a lamp holder fixing seat 302. A high-frequency flashing xenon lamp 304 is fixedly mounted in the lamp holder 303. A forward-extending C-shaped seat 305 is provided at the bottom end of the lamp holder fixing seat 302. A lens 307 is fixedly mounted in the middle of the C-shaped seat 305 via a lens fixing member 306. The lens 307 is located directly in front of the high-frequency flashing xenon lamp 304, and the light transmission position is directly in front of the lens 307. The front of the C-shaped base 305 is equipped with a side turntable connecting plate 308. A connecting bar 309 extends to one side from the top of the side turntable connecting plate 308. A second stepper motor 311, also equipped with an angle sensor, is fixedly mounted to the bottom surface of the connecting bar 309 via a motor mounting plate 310. The output shaft of the second stepper motor 311 is fixedly connected to the center of the outer surface of a side turntable 313 via a connector 312. The side turntable 313 is located in front of the lens 307. Several light source filter apertures 314, for example, eight, are evenly spaced around its circumference. Each of these light source filter apertures 314 is covered with a light source filter 315. The second stepper motor 311 rotates the side turntable 313 at a fixed angle, ensuring that all of the light source filter apertures 314 rotate in the light-transmitting position.

[0076] Taking the setting of 8 light source filter holes 314 as an example, the angular displacement of the second stepper motor 311 when rotating once is set to 45°. Then, when the angle sensor of the second stepper motor 311 measures that the angular displacement of the side turntable 313 reaches 45°, it is recorded as one rotation action of the second stepper motor 311, thereby realizing the fixed angle rotation of the side turntable 313 in units of 45°, and further realizing the 8 light source filter holes 314 staying in the light-transmitting position in turn.

[0077] As a preferred embodiment of the present invention, as shown in Figure 15, a plurality of water-cooling coolers 316 are fixedly attached to the outer surface of the lamp holder 303, and the outer cover of the lamp holder 303 is provided with a protective cover 317 for protecting the water-cooling cooler 316. The rear end of the protective cover 317 is provided with an opening 318, and the coolant circulation pipeline passes through the opening 318 and is connected to the water inlet and outlet of the water-cooling cooler 316.

[0078] As a preferred embodiment of the present invention, as shown in FIG16 , a second mechanical angle limiting mechanism is provided on the light source component 3. The second mechanical angle limiting mechanism includes a second flat spring piece 319, a second micro bearing seat 320, a second micro pin 321, a second roller 322, and a plurality of second roller positioning countersunk holes 323 and a plurality of second roller rolling grooves 324 provided on the rear side of the side turntable 313 and arranged around its center. The inner end of the second flat spring piece 319 is fixedly connected to the front side of the motor mounting plate 310 via a flat spring piece fixing seat 325, the outer end of the second flat spring piece 319 points in a direction away from the detection position, the second micro bearing seat 320 is fixedly provided on the front side of the outer end of the second flat spring piece 319, and the second roller 322 is mounted on the second micro bearing seat 320 via the second micro pin 321. The number of the second roller positioning countersunk holes 323 and the second roller rolling grooves 324 corresponds to the number of the light source filter holes 314, for example, both are 8. The opening position of the second roller positioning countersunk holes 323 corresponds one-to-one to the position of the light source filter holes 314. The opening position of the second roller rolling groove 324 is located between two adjacent second roller positioning countersunk holes 323. The upper plane of the second roller positioning countersunk holes 323 is higher than the upper plane of the second roller rolling groove 324. The recessed second roller rolling groove 324 facilitates the rolling of the second roller 322. The protruding second roller positioning countersunk holes 323 are responsible for providing damping for the second roller 322 and providing a card point for the second roller 322.

[0079] As a preferred embodiment of the present invention, the wheel mounting plate 101 is equipped with an adjustment member for adjusting the perpendicularity between the camera component 2, the wheel component 1, and the objective lens 4. The light source mounting plate 301 is equipped with an adjustment member for adjusting the spacing between the light source component 3 and the wheel component 1. After proper installation and adjustment, the camera component 2, the through hole 111, and the upper beam hole 1072 and lower beam hole 1074 of the three-hole fluorescence filter box 107 located at the detection position are vertically coaxial. The high-frequency flash xenon lamp 304, the lens 307, the light source filter hole 314 located at the light transmission position, and the side beam hole 1075 of the three-hole fluorescence filter box 107 located at the detection position are horizontally coaxial.

[0080] As a preferred embodiment of the present invention, the high-frequency flashing xenon lamp 304 is a light source designed and manufactured using the principle of high-voltage ionized xenon luminescence. The high-frequency flashing xenon lamp 304 is manufactured by encapsulating high-voltage (pressure is about 1 / 10 or less of a continuous xenon lamp) xenon gas in a transparent lampshade, and then inputting a beam of high-frequency pulse voltage signal to both ends of the metal electrodes, so that the xenon gas between the electrodes in the tube is ionized and emits light. The high-frequency flashing xenon lamp 304 of the present invention can provide high-frequency flashing light of 160-7500nm, and its frequency can reach 200Hz. The frequency of the light source used in the prior art is mostly 10Hz. The frequency of the high-frequency flashing xenon lamp 304 of the present invention is 20 times that of the prior art. In comparison, the two can be called high frequency. The single luminous energy of the high-frequency flashing xenon lamp 304 can reach 3 joules, and the continuous luminous time is 5μs. If converted into continuous light, its power output can reach 6×10 5 The high-frequency flash xenon lamp 304 has the characteristics of high luminous intensity, short warm-up time and good stability, and is very suitable for high-speed fluorescence microscopy.

[0081] Referring to FIG17 , the working principle of the present invention is as follows:

[0082] First, the second stepper motor 311 drives the side turntable 313 to rotate a certain angle, rotating the light source filter aperture 314 with the desired light source filter 315 to the light-transmitting position. Once in position, the second roller positioning countersunk hole 323 corresponding to the position of the light source filter aperture 314 contacts the second roller 322 as the side turntable 313 rotates. Simultaneously, the second roller positioning countersunk hole 323 lifts the second roller 322 upward, causing the second flat spring 319 to shift from a straight position to an upward position, locking the second roller 322 with the second roller positioning countersunk hole 323, thereby mechanically positioning the light source filter aperture 314.

[0083] The first stepper motor 108 then drives the mounting shaft 104 to rotate a certain angle, rotating the desired three-hole fluorescent filter box 107 to the detection position. Once in position, the first roller positioning countersunk hole 120 corresponding to the position of the three-hole fluorescent filter box 107 contacts the first roller 119 as the mounting shaft 104 rotates. Simultaneously, the first roller positioning countersunk hole 120 lifts the first roller 119, and the first flat spring 116 then changes from a straight state to an upward state, causing the first roller 119 to lock with the first roller positioning countersunk hole 120, thereby achieving mechanical positioning of the three-hole fluorescent filter box 107.

[0084] Finally, turn on the high-frequency xenon flash lamp 304 and the camera body 201. The high-frequency xenon flash lamp 304 provides high-frequency flash light with a frequency range of 160-7500 nm, with a single burst duration of 5 μs. The light beam from the high-frequency xenon flash lamp 304 passes through lens 307 and light source filter 315 and enters the three-hole fluorescence filter box 107. It is then reflected by mirror 1076 and directed downward through objective lens 4 toward the slide, illuminating the sample on the slide. The light beam is then reflected by the sample and directed upward through objective lens 4, mirror 1076, and fluorescence filter 1073 into the camera body 201, achieving epi-illumination for the high-speed fluorescence microscope.

[0085] The flash xenon lamp technology proposed in the present invention can provide a microsecond-level high-brightness light source without the need for preheating. Without reducing image quality, it can significantly increase the shooting rate (200 frames per second), which is more than 20 times that of existing technologies.

[0086] [Corrected 07.03.2025 in accordance with Rule 26] The foregoing description is merely a preferred embodiment of the present invention and is not intended to limit the present invention. Those skilled in the art will readily appreciate that various modifications and variations of the present invention are possible. Any modifications, equivalent substitutions, or improvements made within the spirit and principles of the present invention shall be included within the scope of protection of the present invention.

Claims

1. A fluorescence microscope illumination method using a flash xenon lamp, characterized in that: Using a high-frequency flash xenon lamp as the light source, the sample on the slide is illuminated by high-frequency flashing through epi-illumination. The specific steps are as follows: Step 1) using a high-frequency xenon flash lamp to emit a lateral flashing light beam; Step 2) filtering the scintillation light beam using a light source filter; Step 3) using a reflector to reflect the scintillation light beam filtered by the light source downward; Step 4) the scintillation light beam reflected downward is irradiated onto a glass slide through an objective lens, wherein a portion of the scintillation light beam is reflected upward after contacting the sample; Step 5) The upward reflected scintillation light beam passes through the objective lens and the reflector in sequence, and then passes through the fluorescence filtering of the fluorescence filter and enters the camera.

2. A fluorescent component based on the fluorescence microscope illumination method according to claim 1, characterized in that: It consists of a wheel component (1), a camera component (2) and a light source component (3); wherein, The wheel disc component (1) comprises a horizontally arranged wheel disc mounting plate (101), a vertically downward central axis (102) is provided on the lower surface of the wheel disc mounting plate (101), a mounting shaft (104) driven by a wheel disc driving mechanism is rotatably sleeved on the central axis (102), and a plurality of three-hole fluorescent filter boxes (107) are evenly arranged on the circumferential surface of the mounting shaft (104) around its axis; a through hole (111) is provided on the wheel disc mounting plate (101), and a detection position is located directly below the through hole (111); the wheel disc driving mechanism drives the mounting shaft (104) to rotate at a fixed angle, thereby realizing that all the three-hole fluorescent filter boxes (107) stay in the detection position in turn; the camera component (2) is located directly above the through hole (111), and the camera component (2), the through hole (111) and the three-hole fluorescent filter box (107) located at the detection position are vertically coaxial; The light source component (3) is located outside the detection position, and the light source component (3) includes a high-frequency flashing xenon lamp (304), a lens (307), and a side turntable (313) driven by a second stepper motor (311). The lens (307) is located directly in front of the high-frequency flashing xenon lamp (304). The position directly in front of the lens (307) is a light-transmitting position. The side turntable (313) is located in front of the lens (307). A plurality of light sources are evenly arranged on the circumference of the disk surface of the side turntable (313). The light filter holes (314) are each covered with a light source filter (315); the second stepping motor (311) drives the side rotating disk (313) to rotate at a fixed angle, so that all the light source filter holes (314) stay in the light transmission position in turn; the high-frequency flashing xenon lamp (304), the lens (307), the light source filter holes (314) located at the light transmission position, and the three-hole fluorescence filter box (107) located at the detection position are transversely coaxial.

3. The fluorescent assembly according to claim 2, wherein: The wheel drive mechanism is composed of a first stepper motor (108), a first spur gear (109), a second spur gear (110) and an angle sensor. The first stepper motor (108) is vertically downwardly mounted on the wheel mounting plate (101) and is located on one side of the central shaft (102) and the camera component (2). The first spur gear (109) is fixedly mounted on the output shaft of the first stepper motor (108). The second spur gear (110) is sleeved on the periphery of the central shaft (102) and is fixedly connected to the upper end surface of the mounting shaft (104). The first spur gear (109) and the second spur gear (110) are at the same height and mesh with each other. The angle sensor includes a magnetic encoder (112), a central magnet (113) and a magnetic encoder (112). , a magnet seat (114) and a socket (115), the magnetic encoder (112) is fixedly arranged on the axis of the lower end surface of the central shaft (102), the magnet seat (114) is fixedly connected to the lower end surface of the mounting shaft (104), the central magnet (113) is fixedly arranged inside the magnet seat (114) and is located directly below the magnetic encoder (112), the cable of the magnetic encoder (112) passes upward through the hollow inner cavity of the central shaft (102) and is connected to the socket (115), the first stepper motor (108) measures the angular displacement of the angle sensor, thereby realizing the fixed angle rotation of the mounting shaft (104), and further realizing the alternating docking of all the three-hole fluorescence filter boxes (107) with the lower end of the through hole (111).

4. The fluorescent assembly according to claim 3, wherein: The wheel disc component (1) is provided with a first mechanical angle limiting mechanism, which comprises a first flat spring piece (116), a first micro bearing seat (117), a first micro pin (118), a first roller (119), and a plurality of first roller positioning countersunk holes (120) and a plurality of first roller rolling grooves (121) provided on the plane circumference of the second spur gear (110); the inner end of the first flat spring piece (116) is fixedly connected to the upper end surface of the central shaft (102), the outer end of the first flat spring piece (116) points in the direction of the detection position, the first micro bearing seat (117) is fixedly arranged on the lower surface of the outer end of the first flat spring piece (116), and the first roller ( 119) is installed on the first micro bearing seat (117) through the first micro pin (118); the number of the first roller positioning countersunk holes (120) and the first roller rolling grooves (121) corresponds to the number of the three-hole fluorescent filter box (107), the opening position of the first roller positioning countersunk holes (120) corresponds to the location of the three-hole fluorescent filter box (107), the opening position of the first roller rolling groove (121) is located between two adjacent first roller positioning countersunk holes (120), the upper plane of the first roller positioning countersunk hole (120) is higher than the upper plane of the first roller rolling groove (121), and is responsible for providing damping and card points for the first roller (119).

5. The fluorescent assembly according to claim 2, wherein: The three-hole fluorescence filter box (107) comprises a square box body (1071), the top surface of the square box body (1071) is provided with a light beam upper through hole (1072) for docking with the lower end of the through hole (111), the light beam upper through hole (1072) is covered with a fluorescence filter (1073), the bottom surface of the square box body (1071) is provided with a light beam lower through hole (1074) for docking with the upper end of the objective lens (4), the light beam upper through hole (1072) is covered with a fluorescence filter (1073), The hole (1072) is concentric with the lower light beam through hole (1074) at the top and bottom. A light beam side through hole (1075) for docking with the inner end of the light source filter hole (314) is provided on one side surface of the square box body (1071). A reflector (1076) is obliquely arranged inside the square box body (1071). The light source component (3) provides epi-illumination to the slide located below the objective lens (4) through the reflector (1076).

6. The fluorescent assembly according to claim 4, characterized in that: The circumferential surface of the installation shaft (104) is uniformly arranged with a plurality of vertically arranged dovetail protrusions (122) around its axis. The number of the dovetail protrusions (122) corresponds to the number of the three-hole fluorescent filter box (107). A dovetail groove (123) capable of plugging and matching with the dovetail protrusion (122) is provided on one side of each of the three-hole fluorescent filter boxes (107). The position of the dovetail groove (123) is opposite to the position of the light beam side through hole (1075). A limiting plate (124) is provided on the outer periphery of the upper end of the installation shaft (104), and the limiting plate (124) is fixedly connected to the upper end faces of all the dovetail protrusions (122). The three-hole fluorescent filter box (107) is connected to the dovetail protrusion (122) through the dovetail groove (123). 22) is matched to realize quick installation with the installation shaft (104), and positioning is realized by the limit plate (124); a C-shaped installation groove (125) is provided on the surface of each dovetail protrusion (122), the notch of the C-shaped installation groove (125) extends to a side of the dovetail protrusion (122), and a slanting pressure block (126) is provided in the C-shaped installation groove (125) and can protrude or retract relative to the outer wall of the installation shaft (104), an edge of the slanting pressure block (126) is located at the notch of the C-shaped installation groove (125) and is flush with the edge of the dovetail protrusion (122), and the dovetail groove (123) is locked or unlocked with the dovetail protrusion (122) by the retraction or protrusion of the slanting pressure block (126).

7. The fluorescent assembly according to claim 2, characterized in that: The light source component (3) comprises a horizontally arranged light source mounting plate (301); a lamp holder seat (303) is provided below the light source mounting plate (301) via a lamp holder fixing seat (302); the high-frequency flash xenon lamp (304) is fixedly mounted in the lamp holder seat (303); a forwardly extending C-shaped seat (305) is provided at the bottom end of the lamp holder fixing seat (302); the lens (307) is fixedly mounted on the C-shaped seat (305) via a lens fixing member (306); 5), a side turntable connecting plate (308) is provided at the front of the C-shaped seat (305), a connecting strip (309) extending to one side is provided at the top of the side turntable connecting plate (308), the second stepper motor (311) is fixedly mounted on the lower surface of the connecting strip (309) through a motor mounting plate (310), and the center of the outer side surface of the side turntable (313) is fixedly connected to the output shaft of the second stepper motor (311) through a connecting piece (312).

8. The fluorescent assembly according to claim 7, characterized in that: A plurality of water-cooling coolers (316) are fixedly attached to the outer surface of the lamp holder (303), and an outer cover of the lamp holder (303) is provided with a protective cover (317) for protecting the water-cooling coolers (316), and an opening (318) is provided at the rear end of the protective cover (317) for the cooling liquid circulation pipeline to pass through.

9. The fluorescent assembly according to claim 7, characterized in that: The light source component (3) is provided with a second mechanical angle limiting mechanism, which comprises a second flat spring piece (319), a second micro bearing seat (320), a second micro pin (321), a second roller (322), and a plurality of second roller positioning countersunk holes (323) and a plurality of second roller rolling grooves (324) which are arranged on the rear side of the side turntable (313) and around the center thereof; the inner end of the second flat spring piece (319) is fixedly connected to the front side of the motor mounting plate (310) via a flat spring piece fixing seat (325); the outer end of the second flat spring piece (319) points in a direction away from the detection position; the second micro bearing seat (320) is fixedly arranged at the outer end of the second flat spring piece (319). The front side of the second roller (322) is mounted on the second micro bearing seat (320) through the second micro pin (321); the number of the second roller positioning countersunk holes (323) and the second roller rolling grooves (324) corresponds to the number of the light source filter holes (314); the opening position of the second roller positioning countersunk holes (323) corresponds to the location of the light source filter holes (314); the opening position of the second roller rolling grooves (324) is located between two adjacent second roller positioning countersunk holes (323); the upper plane of the second roller positioning countersunk holes (323) is higher than the upper plane of the second roller rolling grooves (324), and is responsible for providing damping and a card point for the second roller (322).

10. The fluorescent assembly according to claim 2, characterized in that: The high-frequency flash xenon lamp (304) can provide high-frequency flash light of 160-7500 nm, with a maximum frequency of 200 Hz.

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