A modular deep ultraviolet multi-dimensional laser confocal microscopy device

Abstract

The present invention relates to the field of deep ultraviolet multi-dimensional laser confocal microscopy technology, in particular to a modular deep ultraviolet multi-dimensional laser confocal microscopy device, including a laser confocal scanning microscope, a confocal mainframe connected to the laser confocal scanning microscope; a fluorescence spectrometer and a fluorescence lifetime imaging component are provided on the top of the confocal mainframe, and a device for introducing part of the emitted light into the fluorescence spectrometer and the fluorescence lifetime imaging component through a spectroscope is provided in the confocal mainframe; a light source introduction interface is provided on one side of the confocal mainframe, a multi-channel light source beam combining box is connected to the light source introduction interface, a first continuous laser, a pulsed laser and a second continuous laser are connected to the multi-channel light source beam combining box through an optical fiber, and a spectroscope for combining the multi-channel light sources is provided in the multi-channel light source beam combining box. The present invention not only has the ability to overcome the interference of complex background fluorescence and scattered light, but also ensures high light transmittance in the deep ultraviolet region.

Description

Technical Field

[0001] The invention relates to the technical field of deep ultraviolet multi-dimensional laser confocal microscopy, and in particular to a modular deep ultraviolet multi-dimensional laser confocal microscopy device. Background Art

[0002] Traditional LCSM instruments have a transmittance of deep ultraviolet light close to zero and the sample matrix often has interferences such as strong background signal scattered light. The LCSM technology alone cannot achieve microscopic quantitative analysis of a class of organic pollutants represented by PAHs in complex matrices. Therefore, we proposed a modular deep ultraviolet multi-dimensional laser confocal microscopy device. Summary of the invention

[0003] The purpose of the present invention is to solve the shortcomings of the prior art and to propose a modular deep ultraviolet multi-dimensional laser confocal microscopy device.

[0004] In order to achieve the above object, the present invention adopts the following technical solutions:

[0005] A modular deep ultraviolet multi-dimensional laser confocal microscopy device is designed, comprising a laser confocal scanning microscope, wherein a confocal host is connected to a fluorescence introduction port of the laser confocal scanning microscope;

[0006] The top of the confocal mainframe is provided with a first interface and a second interface, the first interface is connected to a fluorescence spectrometer via a liquid optical fiber, the second interface is connected to a fluorescence lifetime imaging component via a liquid optical fiber, and a device is provided in the confocal mainframe to guide part of the emitted light to the first interface and the second interface via a spectroscope;

[0007] A light source introduction interface is provided on one side of the confocal host, and a multi-way light source combining box is connected to the light source introduction interface. The multi-way light source combining box is provided with a first light input interface, a second light input interface, and a third light input interface. The first light input interface is connected to a first continuous laser via an optical fiber, the second light input interface is connected to a pulsed laser via an optical fiber, and the third light input interface is connected to a second continuous laser via an optical fiber. A spectroscope for combining multiple light sources is provided in the multi-way light source combining box.

[0008] Furthermore, a fixing plate is provided on one side of the light source introduction interface of the confocal mainframe, and mounting seats for connecting the first continuous laser, the pulse laser and the second continuous laser are respectively provided on the fixing plate.

[0009] Furthermore, the multi-path light source beam combining box is provided with a connection port connected to the light source introduction interface, and a light path adjustment mechanism is provided between the connection port and the light source introduction interface.

[0010] Furthermore, the optical path adjustment mechanism includes an adjustment plate and three groups of adjustment units, each group of adjustment units includes a fixed connecting seat, an adjustment bolt and a support spring, the adjustment plate is fixed on the circumference of the connection port, the fixed connecting seat is evenly distributed on the circumference of the light source introduction interface, the adjustment bolt passes through the adjustment plate and is threadedly connected to the fixed connecting seat, the support spring is arranged on the adjustment bolt, and the two ends of the support spring respectively abut against the adjustment plate and the fixed connecting seat.

[0011] Furthermore, a light path calibration mechanism is also provided inside the connection port and the light source introduction interface.

[0012] Furthermore, the optical path calibration mechanism includes a first circuit board and a second circuit board in an annular structure, a signal transmitter is provided on the circumference of the first circuit board and below each adjustment unit, a signal receiver is provided on the circumference of the second circuit board and below each adjustment unit, and the confocal host is provided with a display screen for displaying the correction alignment of the signal transmitter and the signal receiver.

[0013] Furthermore, the first continuous laser is a 266nm continuous ultraviolet laser.

[0014] Furthermore, the pulse laser is a 266nm pulsed ultraviolet laser.

[0015] Furthermore, the second continuous laser is a 355nm continuous ultraviolet laser.

[0016] The modular deep ultraviolet multi-dimensional laser confocal microscopy device proposed by the present invention has the following beneficial effects: the present invention adopts modular design, which is convenient for the installation and assembly of the device, and can ensure the acquisition of clear fluorescence images of fluorescent substances in the sample micro-region; the four-dimensional fluorescence spectrum of excitation-emission-nanosecond time resolution-fluorescence intensity of fluorescent substances in the sample micro-region can be quickly obtained. At the same time, all functions of mainstream confocal can be achieved at ultraviolet excitation wavelengths of 266nm and 355nm, and it not only has the ability to overcome the interference of complex background fluorescence and scattered light, but also ensures high transmittance in the deep ultraviolet region (230-350mm), so as to microscopically and quantitatively analyze the migration and transformation of organic pollutants represented by PAHs on typical environmental micro-interfaces. BRIEF DESCRIPTION OF THE DRAWINGS

[0017] The accompanying drawings are used to provide a further understanding of the present invention and constitute a part of the specification. Together with the embodiments of the present invention, they are used to explain the present invention and do not constitute a limitation of the present invention. In the accompanying drawings:

[0018] Figure 1 It is a three-dimensional structural schematic diagram of the present invention;

[0019] Figure 2 It is a front structural schematic diagram of the present invention;

[0020] Figure 3 yes Figure 2 An enlarged schematic diagram of point A in FIG.

[0021] Figure 4 yes Figure 2 Schematic diagram of the internal structure about point A;

[0022] Marked in the figure are: laser confocal scanning microscope 1, confocal host 2, light source introduction interface 21, first interface 22, second interface 23, fluorescence spectrometer 4, fluorescence lifetime imaging component 5, fixing plate 6, mounting seat 61, multi-channel light source combining box 7, first light input interface 71, second light input interface 72, third light input interface 73, connecting port 74, first continuous laser 81, pulse laser 82, second continuous laser 83, optical path adjustment mechanism 9, adjustment plate 91, fixed connecting seat 92, adjustment bolt 93, support springs 94, 95, first circuit board 101, second circuit board 102, signal transmitter 103, signal receiver 104, display screen 105. DETAILED DESCRIPTION

[0023] The present invention is further described below in conjunction with specific embodiments. These embodiments are only used to illustrate the present invention and are not used to limit the scope of the present invention. In the description of the present invention, it should be noted that, unless otherwise clearly specified and limited, the terms "installed", "provided with", and "connected" should be understood in a broad sense. For example, it can be a fixed connection, a detachable connection, or an integral connection; it can be a mechanical connection or an electrical connection; it can be directly connected, or indirectly connected through an intermediate medium, or it can be internal communication between two components. For ordinary technicians in this field, the specific meanings of the above terms in the present invention can be understood according to specific circumstances.

[0024] The structural features of the present invention are now described in detail with reference to the accompanying drawings.

[0025] See also Figure 1-4 A modular deep ultraviolet multi-dimensional laser confocal microscopy device includes a laser confocal scanning microscope 1. A confocal host 2 is connected to the fluorescence introduction port of the laser confocal scanning microscope 1. The laser confocal scanning microscope 1 adopts an FV3000 laser confocal scanning microscope. When replacing the confocal host 2, two groups of lenses and objective lens groups are required for the incident light path. The lens adopts an ultraviolet lens group, and the objective lens group adopts an ultraviolet transmitting objective lens. The ultraviolet transmitting objective lenses are LUM-UVB ultraviolet transmitting objective lens and MPALNUV ultraviolet transmitting objective lens respectively.

[0026] A first interface 22 and a second interface 23 are provided on the top of the confocal host 2. The first interface 22 is connected to a fluorescence spectrometer 4 via a liquid optical fiber. The second interface 23 is connected to a fluorescence lifetime imaging component 5 via a liquid optical fiber. The fluorescence lifetime imaging component 5 adopts a FLIM detector. A device is provided in the confocal host 2 to introduce part of the emitted light into the first interface 22 and the second interface 23 via a spectroscope. The fluorescence lifetime imaging component 5 adopts a PDL828 fluorescence lifetime imaging component, and the fluorescence spectrometer 4 adopts a fluorescence spectrometer FLS1000.

[0027] The confocal host 2 is provided with a FLIM system optical path. The FLIM system optical path enters and exits the confocal system after beam combining by a beam splitter prism, passes through a scanning head and optical components, and reaches the sample. After the sample is excited, it passes through optical components and has an upper outlet, and reaches the FLIM detector and spectrometer respectively through a beam splitter.

[0028] A light source introduction interface 21 is provided on one side of the confocal host 2, and a multi-way light source combining box 7 is connected to the light source introduction interface 21. The multi-way light source combining box 7 is provided with a first light input interface 71, a second light input interface 72, and a third light input interface 73. The first light input interface 71 is connected to a first continuous laser 81 through an optical fiber, the second light input interface 72 is connected to a pulse laser 82 through an optical fiber, and the third light input interface 73 is connected to a second continuous laser 83 through an optical fiber. A spectrometer for combining multiple light sources is provided in the multi-way light source combining box 7. The first continuous laser 81 is a 266nm continuous ultraviolet laser, the pulse laser 82 is a 266nm pulsed ultraviolet laser, and the second continuous laser 83 is a 355nm continuous ultraviolet laser.

[0029] The confocal host 2 is equipped with a confocal system, which is controlled by the original confocal software. The fluorescence lifetime imaging system of the fluorescence lifetime imaging component 5 also has its own software and can also be controlled by the Olympus external triggering function. The three lasers, the first continuous laser 81, the pulse laser 82 and the second continuous laser 83, can be linearly adjusted on the PC for light intensity, and the overall system runs smoothly without being too cumbersome.

[0030] The modular deep ultraviolet multi-dimensional laser confocal microscopy device of the present invention adopts a modular design, which is convenient for the installation and assembly of the device, and can ensure the acquisition of a clear fluorescent image of the fluorescent substance in the sample micro-region; the four-dimensional fluorescence spectrum of excitation-emission-nanosecond time resolution-fluorescence intensity of the fluorescent substance in the sample micro-region can be quickly obtained. At the same time, all the functions of mainstream confocal can be achieved at 266nm and 355nm ultraviolet excitation wavelengths, and it not only has the ability to overcome the interference of complex background fluorescence and scattered light, but also ensures high transmittance in the deep ultraviolet region (230-350nm), and can further microscopically and quantitatively analyze the migration and transformation of organic pollutants represented by PAHs on typical environmental micro-interfaces.

[0031] See also Figure 3 and 4 Furthermore, a fixing plate 6 is provided on one side of the light source introduction interface 21 of the confocal main unit 2, and mounting seats 61 for connecting the first continuous laser 81, the pulse laser 82 and the second continuous laser 83 are provided on the fixing plate 6.

[0032] A connection port 74 is provided on the multi-channel light source combining box 7 for docking with the light source introduction interface 21, and an optical path adjustment mechanism 9 is provided between the connection port 74 and the light source introduction interface 21; the optical path adjustment mechanism 9 includes an adjustment plate 91 and three groups of adjustment units, each group of adjustment units includes a fixed connection seat 92, an adjustment bolt 93 and a support spring 94, the adjustment plate 91 is fixed on the circumference of the connection port 74, the fixed connection seats 92 are evenly distributed on the circumference of the light source introduction interface 21, the adjustment bolt 93 passes through the adjustment plate 91 and is threadedly connected to the fixed connection seat 92, the support spring 94 is arranged on the adjustment bolt 93, and the two ends of the support spring 94 respectively abut against the adjustment plate 91 and the fixed connection seat 92.

[0033] An optical path calibration mechanism is also provided inside the connection port 74 and the light source introduction interface 21; the optical path calibration mechanism includes a first circuit board 101 and a second circuit board 102 of an annular structure, a signal transmitter 103 is provided on the circumference of the first circuit board 101 and below each adjustment unit, a signal receiver 104 is provided on the circumference of the second circuit board 102 and below each adjustment unit, and a display screen 105 for displaying the correction alignment of the signal transmitter 103 and the signal receiver 104 is provided on the confocal host 2.

[0034] The above description is only a preferred embodiment of the present invention and is not intended to limit the present invention. Although the present invention is described in detail with reference to the aforementioned embodiments, those skilled in the art can still modify the technical solutions described in the aforementioned embodiments or replace some of the technical features therein by equivalents. 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 modular deep ultraviolet multi-dimensional laser confocal microscopy device, comprising a laser confocal scanning microscope (1), characterized in that: The fluorescence introduction port of the laser confocal scanning microscope (1) is connected to a confocal host (2); The top of the confocal mainframe (2) is provided with a first interface (22) and a second interface (23); the first interface (22) is connected to a fluorescence spectrometer (4) via a liquid optical fiber; the second interface (23) is connected to a fluorescence lifetime imaging component (5) via a liquid optical fiber; and a device is provided inside the confocal mainframe (2) for introducing part of the emitted light into the first interface (22) and the second interface (23) via a spectroscope; A light source introduction interface (21) is provided on one side of the confocal mainframe (2); a multi-path light source beam combining box (7) is connected to the light source introduction interface (21); a first light input interface (71), a second light input interface (72), and a third light input interface (73) are provided on the multi-path light source beam combining box (7); the first light input interface (71) is connected to a first continuous laser (81) via an optical fiber; the second light input interface (72) is connected to a pulse laser (82) via an optical fiber; the third light input interface (73) is connected to a second continuous laser (83) via an optical fiber; and a beam splitter for combining the multi-path light sources is provided in the multi-path light source beam combining box (7); The confocal mainframe (2) is provided with a fixing plate (6) on one side of the light source introduction interface (21), and mounting seats (61) for connecting the first continuous laser (81), the pulse laser (82) and the second continuous laser (83) are respectively provided on the fixing plate (6); The multi-path light source beam combining box (7) is provided with a connection port (74) connected to the light source introduction interface (21), and a light path adjustment mechanism (9) is provided between the connection port (74) and the light source introduction interface (21); The first continuous laser (81) is a 266nm continuous ultraviolet laser; The pulse laser (82) is a 266nm pulsed ultraviolet laser; The second continuous laser (83) is a 355nm continuous ultraviolet laser.

2. A modular deep ultraviolet multi-dimensional laser confocal microscopy device according to claim 1, characterized in that: The optical path adjustment mechanism (9) comprises an adjustment plate (91) and three groups of adjustment units, each group of adjustment units comprises a fixed connection seat (92), an adjustment bolt (93) and a support spring (94), the adjustment plate (91) is fixed on the circumference of the connection port (74), the fixed connection seat (92) is evenly distributed on the circumference of the light source introduction interface (21), the adjustment bolt (93) passes through the adjustment plate (91) and is threadedly connected to the fixed connection seat (92), the support spring (94) is arranged on the adjustment bolt (93), and the two ends of the support spring (94) respectively contact the adjustment plate (91) and the fixed connection seat (92).

3. A modular deep ultraviolet multi-dimensional laser confocal microscopy device according to claim 2, characterized in that: An optical path calibration mechanism is also provided inside the connection port (74) and the light source introduction interface (21).

4. A modular deep ultraviolet multi-dimensional laser confocal microscopy device according to claim 3, characterized in that: The optical path calibration mechanism comprises a first circuit board (101) and a second circuit board (102) of an annular structure; a signal transmitter (103) is provided on the circumference of the first circuit board (101) and below each adjustment unit; a signal receiver (104) is provided on the circumference of the second circuit board (102) and below each adjustment unit; and a display screen (105) for displaying the alignment correction of the signal transmitter (103) and the signal receiver (104) is provided on the confocal main unit (2).

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