CN211927700U - Visual background device - Google Patents
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- CN211927700U CN211927700U CN201921449940.3U CN201921449940U CN211927700U CN 211927700 U CN211927700 U CN 211927700U CN 201921449940 U CN201921449940 U CN 201921449940U CN 211927700 U CN211927700 U CN 211927700U
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Abstract
The embodiment of the application provides a visual background device, which is used for a fluorescence microscopic optical system of a fluorescence staining cell scanning and analyzing system; one side of the visual background device can provide fluorescence as background light of an observed object, and one side of the visual background device capable of providing the fluorescence is provided with a non-reflection area which passes through or absorbs excitation light; wherein the non-reflective region is configured to face an objective lens of the fluorescence microscopy optical system to reduce reflection of excitation light by the visual background device, the excitation light being excitation light transmitted through the objective lens of the fluorescence microscopy optical system. The technical problem that the visual background device reflects exciting light is solved.
Description
Technical Field
The application relates to the technical field of fluorescence microscopy, in particular to a visual background device.
Background
A traditional fluorescence microscopic optical system in a fluorescence staining cell scanning and analyzing system adopts a passive fluorescence lining plate as a visual background of an observed object, namely the passive fluorescence lining plate is used as a visual background device of the fluorescence microscopic optical system. As shown in fig. 1, the excitation light 11 transmitted through the objective lens 10 excites the object to be observed 20 to generate fluorescence, and the objective lens 10 detects the fluorescence to study various living things, wherein the objective lens 10 is an epi-fluorescence microscope objective lens. To identify the edge of the observed object, a passive fluorescent mount 30 is typically provided behind the observed object, the passive fluorescent mount 30 having fluorescent particles thereon that emit fluorescent light that serves as a visual background light source for illuminating the observed object 20. The passive fluorescent lining plate 30 can reflect the exciting light 11 to generate a reflection phenomenon, and a halo can be generated on the surface of an observed object during microscopic imaging, so that the quality of the fluorescent microscopic imaging is reduced.
Therefore, the reflection of the excitation light by the visual background device is a technical problem which needs to be solved urgently by those skilled in the art.
The above information disclosed in the background section is only for enhancement of understanding of the background of the present application and therefore it may contain information that does not form the prior art that is known to those of ordinary skill in the art.
SUMMERY OF THE UTILITY MODEL
The embodiment of the application provides a visual background device to solve the technical problem that the visual background device reflects exciting light.
The embodiment of the application provides a visual background device which is used for a fluorescent staining cell scanning and analyzing system. One side of the visual background device can provide fluorescence as background light of an observed object, and one side of the visual background device capable of providing the fluorescence is provided with a non-reflection area which passes through or absorbs excitation light;
wherein the non-reflective region is configured to face an objective lens of the fluorescence microscopy optical system to reduce reflection of excitation light by the visual background device, the excitation light being excitation light transmitted through the objective lens of the fluorescence microscopy optical system.
Due to the adoption of the technical scheme, the embodiment of the application has the following technical effects:
the side of the visual background device capable of providing fluorescence has a non-reflective region that does not reflect the excitation light, but passes or absorbs it. In this way, the visual background means reflect no or less excitation light due to the presence of the non-reflective areas. Compared with the background technology, the visual background device of the fluorescence microscopic optical system provided by the embodiment of the application has the advantages that the reflection of exciting light is less, the reflection phenomenon is less, the fluorescence microscopic optical system can not generate a halo on the surface of an observed object during microscopic imaging, and the quality of the fluorescence microscopic imaging is improved.
Drawings
The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the application and together with the description serve to explain the application and not to limit the application. In the drawings:
FIG. 1 is a schematic diagram of a conventional fluorescence microscope optical system in the background art using a passive fluorescence lining plate as a visual background of an observed object;
FIG. 2 is a schematic view of a visual background arrangement of a fluorescence microscopy optical system according to an embodiment of the present application;
FIG. 3 is a schematic view of the visual background apparatus and objective lens of the fluorescence microscopy optical system shown in FIG. 2;
FIG. 4 is a schematic view of a fluorescence microscopy optical system according to an embodiment of the present application;
FIG. 5 is a schematic diagram of a PWM dimming device of the fluorescence microscopy optical system shown in FIG. 4;
fig. 6 is a schematic diagram of the pulse voltage output by the PWM controller of the PWM dimming device shown in fig. 5.
Description of reference numerals:
in the background art:
10 objective lens, 11 exciting light, 20 observed object and 30 passive fluorescent lining plate;
in the embodiment of the application:
310 non-reflective area, 320 fluorescent plate, 321 power supply wire, 331 objective lens, 332 observed object,
333PWM dimming means, 334 filtering means,
100LED light source arrangement, 210PWM controller, 220 voltage source.
Detailed Description
In order to make the technical solutions and advantages of the embodiments of the present application more apparent, the following further detailed description of the exemplary embodiments of the present application with reference to the accompanying drawings makes it clear that the described embodiments are only a part of the embodiments of the present application, and are not exhaustive of all embodiments. It should be noted that the embodiments and features of the embodiments in the present application may be combined with each other without conflict.
A fluorescence staining cell scanning and analyzing system, called CTC scanning and analyzing system for short, is a system for scanning and identifying 360-degree images of staining cells attached to a needle-shaped carrier. The fluorescent staining cell scanning and analyzing system comprises a plurality of hardware devices, and the software is loaded with analysis software. The LED light source device in the first embodiment is a visual background device of the fluorescent staining cell scanning and analyzing system, and the fluorescent microscopic optical system in the second embodiment is a fluorescent microscopic optical system of the fluorescent staining cell scanning and analyzing system.
Example one
FIG. 2 is a schematic view of a visual background arrangement of a fluorescence microscopy optical system according to an embodiment of the present application;
fig. 3 is a schematic view of the visual background apparatus and the objective lens of the fluorescence microscopy optical system shown in fig. 2.
As shown in fig. 2 and 3, the visual background device of the fluorescence microscope optical system according to the embodiment of the present application is used for the fluorescence microscope optical system of the fluorescence staining cell scanning and analyzing system. One side of the visual background device can provide fluorescence as background light of the observed object 332, and one side of the visual background device capable of providing fluorescence has a non-reflective region 310, and the non-reflective region 310 passes through or absorbs excitation light;
wherein the non-reflective region 310 is configured to face an objective lens of the fluorescence microscopy optical system to reduce reflection of excitation light by the visual background device, the excitation light being transmitted through the objective lens of the fluorescence microscopy optical system.
The visual background device of the fluorescence microscope optical system of the embodiment of the application has a non-reflection area on one side capable of providing fluorescence, and the non-reflection area does not reflect exciting light but passes or absorbs the exciting light. In this way, the visual background means reflect no or less excitation light due to the presence of the non-reflective areas. Compared with the background technology, the visual background device of the fluorescence microscopic optical system provided by the embodiment of the application has the advantages that the reflection of exciting light is less, the reflection phenomenon is less, the fluorescence microscopic optical system can not generate a halo on the surface of an observed object during microscopic imaging, and the quality of the fluorescence microscopic imaging is improved.
In practice, as shown in fig. 2 and 3, the non-reflective region 310 is a hollow region extending through the thickness of the visual background device.
In this way, the hollow region acts as a non-reflective region through which excitation light can pass directly; meanwhile, the cost of the visual background device is low.
In an embodiment, the outer contour of the side of the visual background means providing fluorescence has a size larger than the diameter of the field of view of the objective lens.
Thus, the visual background device can provide fluorescence for the whole field range of the objective lens and improve the brightness of the field of the objective lens.
In an implementation, the visual background means may be a circular or rectangular frame visual background means.
In this way, the annular or rectangular frame visual background device, the hollow part is used as a non-reflection area, one side of the solid part can provide fluorescence, and the fluorescence of the whole field range of the objective lens is uniform.
In an implementation, as an alternative, as shown in fig. 2 and 3, the visual background device includes:
two symmetrically arranged fluorescent plates 320, one side of which can provide fluorescence, the light emitting side of each fluorescent plate faces to the same side, and two fluorescent plates are arranged at intervals to serve as the hollow area of the visual background device.
The visual background device with the structure has a simple structure and is convenient to process and manufacture.
In implementation, the fluorescent plates are fluorescent plates of monochromatic light sources, and each fluorescent plate is connected with a power supply through a power supply lead 321 and a circuit switch;
the circuit switch is used for controlling the power on-off of the fluorescent plate so as to control the existence of fluorescence of the visual background device.
The fluorescent plate is an active fluorescent plate, firstly, the intensity of the emitted fluorescence is relatively stable, and the imaging effect of a fluorescence microscopic optical system can be relatively stable during microscopic imaging; secondly, the existence of fluorescence of the visual background device can be flexibly controlled, and the device is more flexible; thirdly, the wavelength and the intensity of the fluorescence provided by the fluorescent plate can be flexibly selected according to actual needs.
In practice, as shown in fig. 2 and 3, the phosphor plate 320 is a rectangular phosphor plate.
The rectangular fluorescent plate is simple in shape and convenient to process and manufacture.
In practice, as shown in fig. 3, the width of the hollow area between two of the fluorescent plates 320 satisfies the following relation:
a＞2×s×tanβ；
wherein a is a width of a hollow region between the two fluorescent plates, s is a distance between the objective lens and the visual background device, and β is a divergence angle of the excitation light transmitted through the objective lens.
As shown in fig. 3, s is m + n, n is the distance from the observed object 332 to the objective lens, and m is the distance from the observed object 332 to the side of the visual background device capable of providing fluorescence; or n is the distance from the marker to the objective lens, m is the distance from the marker to the side of the visual background device capable of providing fluorescence, and the distance between the marker and the observed object is fixed.
β is a divergence angle of the excitation light transmitted through the objective lens, and a value of β is determined after the frequencies of the objective lens and the excitation light are determined. The derivation of a > 2 × s × tan β is as follows:
as shown in fig. 3, in Δ XYZ, according to the geometric relationship,
since YZ is the sum of s,
it can be deduced that a > 2 XS tan β.
In practice, the length C of the phosphor plate1The distance between the outer edges of the long sides of the two fluorescent plates is larger than the diameter of the field of view of the objective lens.
The length of the fluorescent plate and the distance between the outer edges of the long edges of the two fluorescent plates are both larger than the diameter of the field of view of the objective lens, and the fluorescence of the field of view of the whole objective lens is uniform.
As an alternative, the length C of the phosphor plate1Is 1 mm larger than the diameter of the field of view of the objective lens.
As an alternative, the width C of the phosphor plate2Greater than or equal to 0.1 mm.
In an implementation, the fluorescent plate, the filter device of the fluorescence microscope optical system and the fluorescence camera satisfy the following relations:
＜λ(f0)×E0＜K；
wherein f is0Frequency of fluorescence provided to said phosphor plate, E0Is a frequency of f0Energy of fluorescence of (2), λ (f)0) For the filter of the fluorescence microscope optical system to a frequency f0The responsivity of the fluorescence of (a) is the minimum sensitivity of the fluorescence camera of the fluorescence microscopy optical system, and K is the maximum sensitivity of the fluorescence camera of the fluorescence microscopy optical system.
λ(f0)×E0Is the energy of fluorescence, < lambda (f)0)×E0And < K is the energy for expressing fluorescence in the light sensitive range of the fluorescence camera.
Example two
FIG. 4 is a schematic view of a fluorescence microscopy optical system according to an embodiment of the present application; FIG. 5 is a schematic diagram of a PWM dimming device of the fluorescence microscopy optical system shown in FIG. 4; fig. 6 is a schematic diagram of the pulse voltage output by the PWM controller of the PWM dimming device shown in fig. 5.
As shown in fig. 4, the fluorescence microscopy optical system of the embodiment of the present application, which is used for a fluorescence-stained cell scanning and analyzing system, includes the visual background device of the first embodiment.
In an implementation, as shown in fig. 4, the fluorescence microscope optical system further includes an objective 331, a PWM dimming device 333, and a filter 334. The PWM dimming device will be explained below.
As shown in fig. 5 and 6, the PWM dimming device includes:
a voltage source 220;
a PWM controller 210 for controlling on/off of the voltage source to output a pulse voltage;
the pulse voltage is applied to the LED light source device 100, and light emitted by the LED light source device 100 passes through the filter 334 of the fluorescence microscopic optical system and then passes through the objective 331 to become excitation light.
The PWM controller is used for controlling the on-off of the voltage source to form pulse voltage and outputting the pulse voltage, namely the pulse voltage loaded on the LED light source device can be controlled through the PWM controller, and the dimming of the LED light source device can be realized by adjusting the pulse voltage. Compared with the background technology, the dimming of the LED light source device by the PWM dimming device is realized by the rapid control of the digital signal of the PWM controller, the adjusting frequency and the adjusting precision are higher, and the reliability is better; meanwhile, the power of the voltage source can be larger, and high-power dimming can be realized; in addition, the cost of the voltage source is low.
In an implementation, as shown in fig. 6, the PWM controller controls a pulse width and a pulse frequency of the pulse voltage to adjust an average brightness of the LED light source device.
The PWM controller can control the pulse width and the pulse frequency of the pulse voltage, so that the average brightness of the LED light source device can be adjusted.
In practice, the voltage source is a constant voltage source. The voltage of the pulse voltage is fixed, the voltage of the pulse voltage is not adjusted, and the average brightness of the LED light source device can be adjusted only by adjusting the pulse width and the pulse frequency.
In practice, as shown in fig. 5, the voltage source 220, the PWM controller 210 and the LED light source apparatus 100 are serially connected in sequence.
The sequential connection enables a PWM controller to control the on-off of the voltage source to output pulse voltage, and the pulse voltage is loaded on the LED light source device.
In an embodiment, the average luminance of the LED light source device satisfies the following relation:
wherein E isLIs the average brightness of the LED light source arrangement,
v is the voltage of the voltage source, R0Is the equivalent resistance of the voltage source,
R1is the equivalent resistance of the LED light source device,
f is the pulse frequency of the pulse voltage, tau is the pulse width of the pulse voltage,
eta is the electro-optic conversion efficiency of the LED light source device,
and delta T is observation time, and when the PWM dimming device is used as the dimming device of the fluorescence microscope optical system, the delta T is smaller than the minimum exposure time of a fluorescence camera of the fluorescence microscope optical system.
the total work W done by the current of the LED light source device is partially the part E of the current which is converted into lightLThe other part is a part E which converts the work of current into heat,
W＝EL+ E. The electro-optic conversion efficiency of the LED light source device is eta,
thus, it can be deduced
Further derive the result
Further, Δ T is eliminated, and finally, the derivation is carried out
In an implementation, the pulse frequency of the pulse voltage satisfies the following relation:
f×ΔT＞100。
the pulse frequency of the pulse voltage according with the relation can ensure the uniformity of the brightness of the LED light source device.
In an implementation, the pulse width of the pulse voltage satisfies the following relation: (ii) a
Wherein is the minimum sensitivity of a fluorescence camera of the fluorescence microscopy optical system; namely, the average brightness of the LED light source device is greater than the minimum sensitivity of a fluorescence camera of the fluorescence microscope optical system, and the fluorescence camera can sense the light emitted by the LED light source device.
EXAMPLE III
A fluorescent-stained cell scanning and analysis system of an embodiment of the present application includes the visual background apparatus of the first embodiment.
Example four
The fluorescent staining cell scanning and analyzing system of the embodiment of the application comprises the fluorescent microscopic optical system of the second embodiment.
In the description of the present application and the embodiments thereof, it is to be understood that the terms "top", "bottom", "height", and the like, indicate orientations or positional relationships based on those shown in the drawings, are merely for convenience in describing the present application and simplifying the description, and do not indicate or imply that the referred device or element must have a specific orientation, be constructed in a specific orientation, and be operated, and thus, should not be construed as limiting the present application.
In this application and its embodiments, unless expressly stated or limited otherwise, the terms "disposed," "mounted," "connected," "secured," and the like are to be construed broadly, e.g., as meaning fixedly connected, detachably connected, or integral to; the connection can be mechanical connection, electrical connection or communication; either directly or indirectly through intervening media, either internally or in any other relationship. The specific meaning of the above terms in the present application can be understood by those of ordinary skill in the art as appropriate.
In this application and its embodiments, unless expressly stated or limited otherwise, the first feature "on" or "under" the second feature may comprise the first and second features being in direct contact, or may comprise the first and second features being in contact, not directly, but via another feature in between. Also, the first feature being "on," "above" and "over" the second feature includes the first feature being directly on and obliquely above the second feature, or merely indicating that the first feature is at a higher level than the second feature. A first feature being "under," "below," and "beneath" a second feature includes the first feature being directly above and obliquely above the second feature, or simply meaning that the first feature is at a lesser level than the second feature.
The above disclosure provides many different embodiments or examples for implementing different structures of the application. The components and arrangements of specific examples are described above to simplify the present disclosure. Of course, they are merely examples and are not intended to limit the present application. Moreover, the present application may repeat reference numerals and/or letters in the various examples, such repetition is for the purpose of simplicity and clarity and does not in itself dictate a relationship between the various embodiments and/or configurations discussed. In addition, examples of various specific processes and materials are provided herein, but one of ordinary skill in the art may recognize applications of other processes and/or use of other materials.
While the preferred embodiments of the present application have been described, additional variations and modifications in those embodiments may occur to those skilled in the art once they learn of the basic inventive concepts. Therefore, it is intended that the appended claims be interpreted as including preferred embodiments and all alterations and modifications as fall within the scope of the application.
It will be apparent to those skilled in the art that various changes and modifications may be made in the present application without departing from the spirit and scope of the application. Thus, if such modifications and variations of the present application fall within the scope of the claims of the present application and their equivalents, the present application is intended to include such modifications and variations as well.
Claims (10)
1. A visual background device is used for a fluorescence microscopic optical system of a fluorescence staining cell scanning and analyzing system; the device is characterized in that one side of the visual background device can provide fluorescence as background light of an observed object, and one side of the visual background device capable of providing the fluorescence is provided with a non-reflection area which passes through or absorbs excitation light;
wherein the non-reflective region is configured to face an objective lens of the fluorescence microscopy optical system to reduce reflection of excitation light by the visual background device, the excitation light being excitation light transmitted through the objective lens of the fluorescence microscopy optical system.
2. The visual background device of claim 1, wherein the non-reflective regions are hollow regions throughout the thickness of the visual background device.
3. The visual background device of claim 2, wherein the outer contour of the side of the visual background device providing fluorescence has a size larger than the diameter of the field of view of the objective lens.
4. The visual background device of claim 3, wherein the visual background device is a circular or rectangular frame visual background device.
5. The visual background apparatus of claim 3, wherein the visual background apparatus comprises:
the fluorescent screen comprises two fluorescent screens which are symmetrically arranged, wherein one side of each fluorescent screen can provide fluorescence, the light emitting sides of the fluorescent screens face to the same side, and the two fluorescent screens are arranged at intervals to serve as hollow areas of the visual background device.
6. The visual background apparatus of claim 5, wherein the phosphor plates are phosphor plates of a monochromatic light source, each of the phosphor plates being connected to a power source via power supply wires and a circuit switch;
the circuit switch is used for controlling the power on-off of the fluorescent plate so as to control the existence of fluorescence of the visual background device.
7. The visual background apparatus of claim 6, wherein the phosphor plate is a rectangular phosphor plate.
8. The visual background apparatus of claim 7, wherein the width of the hollow region between two of the phosphor plates satisfies the following relationship:
a＞2×s×tanβ；
wherein a is a width of a hollow region between the two fluorescent plates, s is a distance between the objective lens and the visual background device, and β is a divergence angle of the excitation light transmitted through the objective lens.
9. The visual background apparatus of claim 8, wherein the length of the phosphor plate is greater than the diameter of the field of view of the objective lens, and the distance between the outer edges of the long sides of the two phosphor plates is greater than the diameter of the field of view of the objective lens.
10. The visual background apparatus of claim 9 wherein the length of the phosphor plate is 1 mm greater than the diameter of the field of view of the objective lens; the width of the fluorescent plate is more than or equal to 0.1 mm;
the fluorescent plate, the filtering device of the fluorescence microscopic optical system and the fluorescence camera satisfy the following relational expression:
＜λ(f0)×E0＜K；
wherein f is0Frequency of fluorescence provided to said phosphor plate, E0Is a frequency of f0Energy of fluorescence of (2), λ (f)0) For the filter of the fluorescence microscope optical system to a frequency f0The responsivity of the fluorescence of (a) is the minimum sensitivity of the fluorescence camera of the fluorescence microscopy optical system, and K is the maximum sensitivity of the fluorescence camera of the fluorescence microscopy optical system.
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| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111239086A (en) * | 2019-08-30 | 2020-06-05 | 北京临近空间飞行器***工程研究所 | Visual background device, fluorescence microscope optical system, scanning analysis system |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN111239086A (en) * | 2019-08-30 | 2020-06-05 | 北京临近空间飞行器***工程研究所 | Visual background device, fluorescence microscope optical system, scanning analysis system |
| CN111239086B (en) * | 2019-08-30 | 2024-04-05 | 北京临近空间飞行器***工程研究所 | Visual background device, fluorescence microscopic optical system and scanning analysis system |
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