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

Abstract

The present invention discloses a fluorescence microscope illumination method and a fluorescence component using a flash xenon lamp, and the fluorescence component includes a wheel component, a camera component and a light source component. The wheel component includes a wheel mounting plate, and a plurality of three-hole fluorescence filter boxes are arranged on the wheel mounting plate through a central axis and a mounting axis; a through hole is arranged on the wheel mounting plate, and a wheel drive mechanism drives the mounting axis to rotate at a fixed angle to realize that the three-hole fluorescence filter boxes stay in turn directly below the through hole; the light source component includes a high-frequency flash xenon lamp, a lens and a side turntable, the lens is located in front of the high-frequency flash xenon lamp, the side turntable is located in front of the lens, and a plurality of light source filter holes are arranged on the side turntable, and a light source filter is arranged in the light source filter hole; the second stepper motor drives the side turntable to rotate at a fixed angle to realize that the light source filter hole stays in turn directly in front of the lens. The present invention can provide a microsecond-level high-brightness light source, greatly improve the shooting rate without reducing the image quality, and has a short warm-up time and good stability.

Description

Technical Field

[0001] The 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 can be used to image biological samples in a variety of ways, such as two-dimensional, three-dimensional, and time series. Fluorescence microscopy can observe the specific structures and physiological functions of microscopic biological systems such as living cells, animals, and bacteria in real time. Therefore, fluorescence microscopy has been widely used in biomedicine, medical diagnosis, and drug development.

[0003] (a) Application in biomedicine;

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

[0005] (2) Application in medical diagnosis;

[0006] 1. Bio-labeling 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 a variety of diseases; 2. Tissue sections: Fluorescence microscopes can efficiently image tissue sections to help medical scientists conduct more detailed research and diagnosis of diseases.

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

[0008] 1. Drug analysis and screening: Fluorescence microscopes can help scientists quickly screen new drugs and determine their efficacy from the molecular to the cellular level; fluorescent labels can be used to analyze the transport and metabolism of drugs, providing assistance in exploring the mechanism of drug action; 2. Toxicology research: Fluorescence microscopes 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 illumination sources. Monochromatic light is obtained through light source filters and illuminated using epi-illumination methods. Sample fluorescence is obtained through fluorescence filters and then sent to the camera for photographing to obtain sample fluorescence information. However, the high-pressure mercury lamps used in current fluorescence microscopes are continuous light sources. Therefore, when sample fluorescence is weak, the camera requires a longer exposure time (100ms or longer), and the corresponding photographing rate cannot be increased. This results in the inability of existing fluorescence microscopy technology to achieve high-speed image scanning and rapid output of large sample volumes. Summary of the invention

[0010] In view of the defects of the prior art, the present invention provides a fluorescence microscope illumination method and a 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 the 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 by means of 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 flashing light beam filtered by the light source downward;

[0016] Step 4) the scintillation beam reflected downward is irradiated onto the glass slide through the objective lens, and a portion of the scintillation 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 fluorescent filtering of the fluorescent filter and enters the camera.

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

[0019] The wheel disc component comprises a horizontally arranged wheel disc mounting plate, the lower surface of the wheel disc mounting plate is provided with a central axis pointing vertically downward, a mounting shaft driven by a wheel disc driving mechanism is rotatably sleeved on the central shaft, and a plurality of three-hole fluorescent filter boxes are evenly arranged on the circumferential surface of the mounting shaft around its axis; a through hole is provided on the wheel disc mounting plate, and a detection position is located directly below the through hole, and the wheel disc driving mechanism drives the mounting shaft to rotate at a fixed angle, so that all the three-hole fluorescent filter boxes stay in the detection position in turn; the camera component is located directly above the through hole, and the camera component, the through hole and the three-hole fluorescent 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, and a plurality 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 is composed of a first stepper motor, a first spur gear, a second spur gear and an angle sensor. The first stepper motor is vertically mounted 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 surface 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 surface of the central shaft. The magnet seat is fixedly connected to the lower end surface 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 measures the angular displacement of the angle sensor, thereby realizing the fixed-angle rotation of the mounting shaft, and then realizing the alternating docking of all the three-hole fluorescent filter boxes with the lower end of the through hole.

[0022] Furthermore, a first mechanical angle limiting mechanism is provided on the wheel disc component, and the first mechanical angle limiting mechanism includes a first flat spring sheet, a first micro-bearing seat, a first micro-pin shaft, a first roller, and a plurality of first roller positioning countersunk holes and a plurality of first roller rolling grooves provided on the plane circumference of the second spur gear; the inner end of the first flat spring sheet is fixedly connected to the upper end surface of the central axis, the outer end of the first flat spring sheet points to 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 sheet, and the first roller is installed on the first micro-bearing seat through the first micro-pin shaft; 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 incident 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, the number of the dovetail protrusions corresponds to the number of the three-hole fluorescent filter boxes, and a dovetail groove capable of plugging and cooperating with the dovetail protrusion is provided on one side of each of the three-hole fluorescent filter boxes, the position of the dovetail groove is opposite to the position of the light beam side through hole, a limiting plate is provided on the outer sleeve of the upper end of the mounting shaft, and the limiting plate is fixedly connected to the upper end faces of all the dovetail protrusions, and the three-hole fluorescent filter box is connected to the dovetail through the dovetail groove. The cooperation of the protrusion realizes quick installation with the mounting shaft, and positioning is achieved through the limit plate; a C-shaped mounting groove is provided on the surface of each dovetail protrusion, the notch of the C-shaped mounting groove extends to a side edge 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, an 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 retracting or protruding the ramp pressure block.

[0025] Furthermore, the camera component is composed of a camera body, a reducing lens and a lens barrel, the lower end of the lens barrel is fixedly connected to a groove provided 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 arranged 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 C-shaped seat extending forward is arranged 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 part, a side turntable connecting plate is arranged in the front part of the C-shaped seat, a connecting strip extending to one side is arranged on 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 part.

[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 sheet, a second micro-bearing seat, a second micro-pin shaft, a second roller, and a plurality of second roller positioning countersunk holes and a plurality of second roller rolling grooves which are opened on the rear side surface of the side turntable and arranged around its center; the inner end of the second flat spring sheet is fixedly connected to the front side surface of the motor mounting plate through a flat spring sheet fixing seat, the outer end of the second flat spring sheet points in a direction away from the detection position, the second micro-bearing seat is fixedly arranged on the front side surface of the outer end of the second flat spring sheet, and the second roller is installed on the second micro-bearing seat through the second micro-pin shaft; 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 holes corresponds one-to-one to the position of the light source filter holes, 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 160-7500 nm, with a frequency of up to 200 Hz.

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

[0031] The present invention replaces the light source of the fluorescence microscope with a flash xenon lamp and adopts a falling illumination method, thereby providing a microsecond-level high-brightness light source. Without reducing the 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 preheating time is short and the stability is good, so high-speed fluorescence microscope scanning can be achieved.

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

[0033] The present invention adopts dual positioning of an angle sensor and a mechanical limit, so that the rotation angles of the fluorescent filter and the light source filter are more precise and can be kept stable.

[0034] The fluorescent filter box of the present invention is assembled with the installation shaft by means of dovetail slot insertion 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 implement it according to the contents of the specification, the following is a detailed description of the preferred embodiments of the present invention in conjunction with the accompanying drawings. The specific implementation of the present invention is given in detail by the following embodiments and their 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] Figure 1 It is a front perspective stereogram of the present invention;

[0038] Figure 2 It is a back perspective stereogram of the present invention;

[0039] Figure 3 It is a bottom perspective stereogram of the present invention;

[0040] Figure 4 A three-dimensional diagram of a roulette wheel component of the present invention;

[0041] Figure 5 A three-dimensional diagram of the wheel disc component of the present invention after the wheel disc mounting plate is removed;

[0042] Figure 6 It is a schematic diagram of the assembly relationship between the three-hole fluorescent filter box and the mounting shaft in the wheel component of the present invention;

[0043] Figure 7 An exploded view of the central axis, the mounting axis and the angle sensor in the wheel disc component of the present invention;

[0044] Figure 8 It is a front perspective view of a three-hole fluorescent filter box in the wheel disc component of the present invention;

[0045] Fig. 9 It is a front perspective view of a three-hole fluorescent filter box in the wheel disc component of the present invention;

[0046] Fig.10 It is a schematic diagram of the assembly relationship between the oblique pressure block and the mounting shaft in the wheel disc component of the present invention;

[0047] Fig.11 It is a structural diagram of the first mechanical angle limiting mechanism in the wheel disc component of the present invention;

[0048] Fig.12 is a three-dimensional diagram of the camera component of the present invention;

[0049] Fig.13 A rear-side perspective view of the light source component of the present invention;

[0050] Fig.14 It is a front side perspective stereogram of the light source component of the present invention;

[0051] Fig.15 This is a schematic diagram of the installation position of the water-cooling cooler in the light source component of the present invention;

[0052] Fig.16 is a structural diagram of a second mechanical angle limiting mechanism in a light source component of the present invention;

[0053] Fig.17 It is a working principle diagram of the present invention. DETAILED DESCRIPTION

[0054] The preferred embodiments of the present invention will be described in detail below in conjunction with the accompanying drawings so that the purpose, features and advantages of the invention can be more clearly understood. It should be understood that the embodiments shown in the accompanying drawings are not intended to limit the scope of the present invention, but are only intended to illustrate the essential spirit of the technical solution of the present invention.

[0055] In the following description, certain specific details are set forth for the purpose of illustrating various disclosed embodiments 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 the present 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, ie, should be interpreted as "including, but not limited to."

[0057] References throughout the specification to "one embodiment" or "an embodiment" indicate 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 the 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] See also Fig.17 As shown, 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 by means of 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 flashing light beam filtered by the light source downward;

[0064] Step 4) the scintillation beam reflected downward is irradiated onto the glass slide through the objective lens, and a portion of the scintillation 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 fluorescent filtering of the fluorescent filter and enters the camera.

[0066] See also Figures 1 to 3 As shown, an electron microscope fluorescence component 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 a preferred embodiment of the present invention, see Figures 4 to 7 As shown, the roulette component 1 includes a horizontally arranged roulette mounting plate 101, and a vertically downward central axis 102 is arranged on the lower surface of the roulette mounting plate 101. A mounting shaft 104 driven by a roulette drive mechanism is rotatably sleeved on the central axis 102 through two bearings 103. A locking nut 105 is arranged at the bottom of the central axis 102, and a bearing cover plate 106 is sleeved on the outer periphery of the locking nut 105, and the bearing cover plate 106 is fixedly connected to the lower end surface of the mounting shaft 104. A plurality of three-hole fluorescent filter boxes 107 are evenly arranged on the circumferential surface of the mounting shaft 104 around its axis, for example, 6 three-hole fluorescent filter boxes 107 can be arranged to form a six-hole roulette. A through hole 111 is opened on the roulette mounting plate 101, and the detection position is directly below the through hole 111. The roulette drive mechanism drives the mounting shaft 104 to rotate at a fixed angle, so that all the three-hole fluorescent filter boxes 107 stay in the detection position in turn.

[0068] As a preferred embodiment of the present invention, see Figures 8 and 9As shown, the three-hole fluorescent 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, the upper beam through hole 1072 is covered with a fluorescent 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 above and below, a side beam 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, and the light source component 3 provides incident illumination to the slide located below the objective lens 4 through the reflector 1076.

[0069] As a preferred embodiment of the present invention, see Figures 4 and 5 As shown, 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 vertically downwardly mounted on the wheel mounting plate 101, and both are located on one side of the central axis 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 axis 102 and fixedly connected to the upper end surface of the mounting axis 104. The first spur gear 109 and the second spur gear 110 are at the same height position and mesh with each other.

[0070] As a preferred embodiment of the present invention, see Figure 7 As shown, 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, and 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 three-hole fluorescent filter box 107 staying in the detection position in turn.

[0071] Taking setting 6 of the three-hole fluorescent filter boxes 107 as an example, the angular displacement of the first stepper motor 108 for one rotation is set to 60°. When the angle sensor measures that the angular displacement of the mounting shaft 104 reaches 60°, it is recorded as one rotation action 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 a preferred embodiment of the present invention, see Fig.11 As shown, in addition to the angle sensor with the magnetic encoder as the core, the present invention also adopts a first mechanical angle limiting mechanism. The first mechanical angle limiting mechanism includes 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 axis 102, the outer end of the first flat spring piece 116 points to 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, for example, both are 6, and the first roller positioning countersunk holes 120 and the first roller rolling grooves 121 are arranged on the first micro bearing seat 117. 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, see Figures 6 to 10As shown, each of the three-hole fluorescent 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 plurality 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 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, and the position of the dovetail groove 123 is opposite to the position of the beam side through hole 1075. A stopper 124 is sleeved on the outer periphery of the upper end of the mounting shaft 104, and the stopper 124 is fixedly connected to the upper end surfaces of all the dovetail protrusions 122. The three-hole fluorescent filter box 107 is quickly mounted on the mounting shaft 104 through the cooperation between the dovetail groove 123 and the dovetail protrusion 122, and positioning is achieved through the stopper 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 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 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 block 126.

[0074] As a preferred embodiment of the present invention, see Fig.12 As shown, the camera component 2 is located directly above the through hole 111. The camera component 2 is composed of a camera body 201, a reducing lens 202 and a lens barrel 203. The lower end of the lens barrel 203 is fixedly connected to the embedding groove provided 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 through the reducing lens 202.

[0075] As a preferred embodiment of the present invention, see Figures 13-16As shown, 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, and a lamp holder seat 303 is arranged below the light source mounting plate 301 through a lamp holder fixing seat 302, and a high-frequency flashing xenon lamp 304 is fixedly installed in the lamp holder seat 303. A C-shaped seat 305 extending forward is arranged at the bottom end of the lamp holder fixing seat 302, and a lens 307 is fixedly installed in the middle of the C-shaped seat 305 through 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. A side turntable connecting plate 308 is arranged in front of the C-shaped seat 305, and a connecting strip 309 extending to one side is arranged on the top of the side turntable connecting plate 308, and a second stepping motor 311 is fixedly installed on the lower surface of the connecting strip 309 through a motor mounting plate 310, and the second stepping motor 311 is also equipped with an angle sensor. The output shaft of the second stepper motor 311 is fixedly connected to the center of the outer side surface of a side rotating disk 313 through a connecting piece 312. The side rotating disk 313 is located in front of the lens 307. A plurality of light source filter holes 314 are evenly opened on the circumference of the disk surface of the side rotating disk 313, for example, 8 light source filter holes 314 are opened. Each of the light source filter holes 314 is covered with a light source filter 315. The second stepper 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.

[0076] Taking setting 8 of the light source filter holes 314 as an example, the angular displacement of the second stepper motor 311 for one rotation is set to 45°. 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, see Fig.15 As shown, a plurality of water-cooling coolers 316 are fixedly attached to the outer surface of the lamp holder holder 303, and the outer cover of the lamp holder holder 303 is provided with a protective cover 317 for protecting the water-cooling coolers 316. An opening 318 is provided at the rear end of the protective cover 317, and a 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, see Fig.16As shown, 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 which are provided on the rear side of the side turntable 313 and around the center of the circle. The inner end of the second flat spring piece 319 is fixedly connected to the front side of the motor mounting plate 310 through a flat spring piece fixing seat 325, the outer end of the second flat spring piece 319 points to 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 installed 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, for example, both are 8. The opening positions of the second roller positioning countersunk holes 323 correspond one-to-one to the positions of the light source filter holes 314. The opening positions of the second roller rolling grooves 324 are 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. The recessed second roller rolling grooves 324 facilitate 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 provided with an adjusting member for adjusting the verticality of the camera component 2, the wheel component 1 and the objective lens 4, and the light source mounting plate 301 is provided with an adjusting member for adjusting the distance between the light source component 3 and the wheel component 1. After correct installation and adjustment, the camera component 2, the through hole 111, and the upper beam through hole 1072 and the lower beam through hole 1074 of the three-hole fluorescent 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 beam side through hole 1075 of the three-hole fluorescent 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 that of a continuous xenon lamp) xenon gas in a transparent lampshade, and then inputting a 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 luminescent. The high-frequency flashing xenon lamp 304 of the present invention can provide high-frequency flashing light of 160~7500 nm, 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 can reach 6×10 5 W. 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] See also Fig.17 As shown, 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, and rotates the light source filter hole 314 with the required light source filter 315 to the light transmission position. After it is in place, the second roller positioning countersunk hole 323 corresponding to the position of the light source filter hole 314 contacts the second roller 322 as the side turntable 313 rotates, and at the same time, the second roller positioning countersunk hole 323 lifts the second roller 322, and the second flat spring sheet 319 immediately changes from a straight state to an upward state, so that the second roller 322 is clamped with the second roller positioning countersunk hole 323, thereby realizing the mechanical positioning of the light source filter hole 314.

[0083] Then, the first stepper motor 108 drives the installation shaft 104 to rotate a certain angle, and the required three-hole fluorescent filter box 107 is rotated to the detection position. After it is in place, 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 installation shaft 104 rotates, and at the same time, the first roller positioning countersunk hole 120 lifts the first roller 119, and the first flat spring sheet 116 immediately changes from a straight state to an upward state, so that the first roller 119 is clamped with the first roller positioning countersunk hole 120, thereby realizing the mechanical positioning of the three-hole fluorescent filter box 107.

[0084] Finally, turn on the high-frequency flash xenon lamp 304 and the camera body 201. The high-frequency flash xenon lamp 304 provides high-frequency flash light of 160-7500 nm, and the single continuous light emission time is 5μs. The light beam emitted by the high-frequency flash xenon lamp 304 passes through the lens 307 and the light source filter 315 and enters the three-hole fluorescent filter box 107, and then is reflected by the reflector 1076 and emitted downward through the objective lens 4 to the slide, illuminating the sample on the slide, and then the light beam is reflected by the sample and emitted upward through the objective lens 4, the reflector 1076, and the fluorescent filter 1073 to the camera body 201, thereby realizing 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. It can greatly increase the shooting rate (200 frames per second) without reducing the image quality, which is more than 20 times that of the existing technology.

[0086] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. For those skilled in the art, the present invention may have various modifications and variations. Any modification, equivalent replacement, improvement, etc. made within the spirit and principle of the present invention shall be included in the protection scope of the present invention.

Claims

1. A fluorescence component for a fluorescence microscope using a flash xenon lamp, characterized in that: It is composed of a wheel component (1), a camera component (2) and a light source component (3); wherein: The wheel component (1) comprises a horizontally arranged wheel mounting plate (101), the lower surface of the wheel mounting plate (101) being provided with a central axis (102) extending vertically downward, a mounting shaft (104) driven by a wheel driving mechanism being rotatably sleeved on the central axis (102), a plurality of three-hole fluorescent filter boxes (107) being evenly arranged on the circumferential surface of the mounting shaft (104) around its axis; a through hole (111) being provided on the wheel mounting plate (101), a detection position being located directly below the through hole (111), the wheel driving mechanism driving the mounting shaft (104) to rotate at a fixed angle, thereby enabling all the three-hole fluorescent filter boxes (107) to stay in the detection position in turn; the camera component (2) being located directly above the through hole (111), the camera component (2), the through hole (111) and the three-hole fluorescent filter box (107) located at the detection position being vertically coaxial; The light source component (3) is located outside the detection position, and comprises 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); and a plurality of light sources are evenly arranged on the circumference of the disk surface of the side turntable (313). light filter holes (314), each of the light source light filter holes (314) being 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 light filter holes (314) stay in the light transmission position in turn; the high-frequency flashing xenon lamp (304), the lens (307), the light source light filter holes (314) located at the light transmission position, and the three-hole fluorescent filter box (107) located at the detection position are transversely coaxial; 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 mounted vertically downward 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 located at the same height and mesh with each other; the angle sensor comprises 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 the fixed-angle rotation of the mounting shaft (104), and further realizing the alternating docking of all the three-hole fluorescent filter boxes (107) with the lower end of the through hole (111); The wheel disc component (1) is provided with a first mechanical angle limiting mechanism, the first mechanical angle limiting mechanism comprising a first flat spring sheet (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 sheet (116) is fixedly connected to the upper end surface of the central shaft (102), the outer end of the first flat spring sheet (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 sheet (116), and the first roller (119) is fixedly connected to the upper end surface of the central shaft (102). 119) is mounted 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 a card point for the first roller (119); The light source component (3) is provided with a second mechanical angle limiting mechanism, the second mechanical angle limiting mechanism comprising a second flat spring sheet (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 provided on the rear side surface of the side turntable (313) and arranged around the center of the circle; the inner end of the second flat spring sheet (319) is fixedly connected to the front side surface of the motor mounting plate (310) via a flat spring sheet fixing seat (325); the outer end of the second flat spring sheet (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 sheet (319) On the front side, 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); 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 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), and is responsible for providing damping and a clamping point for the second roller (322).

2. The fluorescence component of a fluorescence microscope using a flash xenon lamp according to claim 1, characterized in that: The three-hole fluorescent filter box (107) comprises a square box body (1071), the top surface of the square box body (1071) is provided with an upper light beam through hole (1072) for docking with the lower end of the through hole (111), the upper light beam through hole (1072) is covered with a fluorescent filter (1073), the bottom surface of the square box body (1071) is provided with a lower light beam through hole (1074) for docking with the upper end of the objective lens (4), the upper light beam through hole (1072) is covered with a fluorescent 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 incident illumination to a glass slide located below the objective lens (4) through the reflector (1076).

3. The fluorescence component of a fluorescence microscope using a flash xenon lamp according to claim 2, characterized in that: The circumferential surface of the installation shaft (104) is evenly arranged around its axis with a plurality of vertically arranged dovetail protrusions (122), the number of the dovetail protrusions (122) corresponding to the number of the three-hole fluorescent filter boxes (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 sleeved 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 surfaces of all the dovetail protrusions (122), and the three-hole fluorescent filter box (107) is connected to the dovetail protrusion (122) through the dovetail groove (123). 2) to achieve quick installation with the installation shaft (104), and positioning is achieved through 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 edge of the dovetail protrusion (122), and an inclined pressure block (126) is provided in the C-shaped installation groove (125) and can protrude outward or retract relative to the outer wall of the installation shaft (104), an edge of the inclined 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 inclined pressure block (126).

4. The fluorescence component of a fluorescence microscope using a flash xenon lamp according to claim 1, characterized in that: The light source component (3) comprises a light source mounting plate (301) arranged horizontally, a lamp holder seat (303) being arranged below the light source mounting plate (301) via a lamp holder fixing seat (302), the high-frequency flashing xenon lamp (304) being fixedly mounted in the lamp holder seat (303); a C-shaped seat (305) extending forward is arranged at the bottom end of the lamp holder fixing seat (302), the lens (307) being fixedly mounted on the C-shaped seat (307) via a lens fixing member (306). 5), a side turntable connecting plate (308) is arranged at the front of the C-shaped seat (305), a connecting strip (309) extending to one side is arranged 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) via 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) via a connecting piece (312).

5. The fluorescence component of a fluorescence microscope using a flash xenon lamp according to claim 4, characterized in that: A plurality of water-cooling coolers (316) are fixedly attached to the outer surface of the lamp holder seat (303), and an outer cover of the lamp holder seat (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 facilitating the passage of a cooling liquid circulation pipeline.

6. The fluorescence assembly for a fluorescence microscope using a xenon flash lamp according to claim 1, characterized in that: The high-frequency flashing xenon lamp (304) can provide high-frequency flashing light of 160-7500 nm, with a maximum frequency of 200 Hz.

7. A method for illuminating a fluorescent component of a fluorescence microscope using a flash xenon lamp as claimed in any one of claims 1 to 6, characterized in that: A high-frequency flash xenon lamp is used as the light source, and the sample on the slide is illuminated with high-frequency flashing by 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 flashing light beam filtered by the light source downward; Step 4) the scintillation beam reflected downward is irradiated onto the glass slide through the objective lens, and a portion of the scintillation 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 fluorescent filtering of the fluorescent filter and enters the camera.

Images