CN214584911U

CN214584911U - Detection device

Info

Publication number: CN214584911U
Application number: CN202120638158.7U
Authority: CN
Inventors: 马炯; 糜岚; 费义艳; 龙相安; 陈丽雯
Original assignee: Fudan University; Zhuhai Fudan Innovation Research Institute
Current assignee: Fudan University; Zhuhai Fudan Innovation Research Institute
Priority date: 2021-03-29
Filing date: 2021-03-29
Publication date: 2021-11-02
Anticipated expiration: 2031-03-29

Abstract

The utility model provides a detection device, among the detection device, with first light filter and second light filter integration in the light filter, coordinate the position of switching mechanism adjustment first light filter and second light filter on the light path through the controller to let photoelectric detector obtain first fluorescence signal and second fluorescence signal, make the light path save a light shade and a photoelectric detector at least, be favorable to simplifying the light path, reduce the cost of manufacture.

Description

Detection device

Technical Field

The embodiment of the utility model provides a relate to the optical detection field, especially relate to a detection device.

Background

Optical methods are commonly used to detect whether a lesion occurs in a body organ, and fluorescence lifetime is one of the optical parameters used to detect a lesion.

When a certain substance is excited and transits to a certain excited state, and then returns to the ground state in the form of radiative transition, the average residence time at the upper energy level is the fluorescence lifetime. Referring to fig. 1, a fluorescence lifetime diagram is shown with time (t) on the abscissa and fluorescence intensity (I) on the ordinate. As shown in the schematic diagram, the fluorescence intensity of the molecule is reduced to the maximum value I_maxThe time required for 1/e of (1) was taken as the fluorescence lifetime.

Fluorescence lifetime can be described by the single e index formula:

I(t)＝I_maxexp(-t/τ)

where τ is the fluorescence lifetime, I_maxThe maximum fluorescence intensity value emitted, i (t), is the fluorescence intensity at time t.

The fluorescence lifetime of the fluorescent substance is related to the conditions such as the structure of the fluorescent substance, the polarity of the microenvironment, the viscosity, the pH value and the like, so that the measurement of the fluorescence lifetime can reflect the change of the system, the fluorescence lifetime is mostly in the nanosecond level, and various complex intermolecular interaction processes can be observed through the fluorescence technology.

When the human organ is diseased, the metabolism of the microenvironment where the human cells or tissues are located is abnormal, so that the fluorescence life of certain molecules of the human cells or tissues is influenced.

However, the existing optical detection devices have insufficient accuracy and sensitivity, have a certain rate of missed diagnosis, and usually require a detection device to be placed in a diseased region or a patient tissue sample to be obtained through an operation or biopsy, so that the detection is traumatic to the human body; in addition, the optical path of the existing optical detection device is complex, and a plurality of optical devices are correspondingly used, which is not beneficial to reducing the manufacturing cost of the optical detection device.

Disclosure of Invention

The embodiment of the utility model provides a problem solved provides a detection device, optimizes the light path, reduces the cost of manufacture.

In order to solve the above problem, an embodiment of the present invention provides a detection device, including: a light emitter for providing excitation light, wherein the excitation light projected on the detection object can generate fluorescence; the optical filter comprises a first optical filter, a second optical filter and a switching mechanism, wherein the switching mechanism is used for adjusting the positions of the first optical filter and the second optical filter so that the first optical filter or the second optical filter is positioned on the light path to transmit fluorescence; the fluorescence penetrates through the first optical filter to form first fluorescence, and the fluorescence penetrates through the second optical filter to form second fluorescence; a photodetector for detecting the first fluorescence or the second fluorescence; the controller is connected with the photoelectric detector and the switching mechanism and used for enabling the photoelectric detector to detect first fluorescence to obtain a first fluorescence signal when the switching mechanism controls the first optical filter to be in the position of the optical path; the switching mechanism is further used for enabling the photoelectric detector to obtain a second fluorescence signal when the photoelectric detector detects a second fluorescence when the switching mechanism controls the second optical filter to be in the optical path position; and the time counter is used for obtaining first fluorescence lifetime data based on time according to the first fluorescence signal and is also used for obtaining second fluorescence lifetime data based on time according to the second fluorescence signal.

Optionally, the switching mechanism includes: a carrier; the rotating support is rotatably arranged on the bearing frame, the first optical filter and the second optical filter are positioned on the rotating support, and the planes of the first optical filter and the second optical filter are both vertical to the light path; the controller is used for driving the rotary bearing to rotate relative to the bearing frame, and a rotating plane of the rotary bearing is perpendicular to the light path, so that the first light filter and the second light filter are respectively located at the light path position.

Optionally, the rotation bearing is the ring, the rotation center of rotation bearing does the centre of a circle of rotation bearing, crosses the straight line of the centre of a circle of rotation bearing will the rotation bearing divide into first region and second region, first light filter is located in the first region, the second light filter is located in the second region.

Optionally, the distance from the position where the fluorescence penetrates through the first optical filter or the second optical filter to the circle center is one third to two thirds of the radius of the circular ring.

Optionally, the first optical filter and the second optical filter are both one, the first optical filter and the second optical filter are spaced apart, and the first optical filter and the second optical filter are both circular.

Optionally, the frame of the rotary bearing is a circle, the rotation center of the rotary bearing is the circle center of the rotary bearing, the first optical filter and the second optical filter are located at two sides of the circle center of the rotary bearing, and the circle center of the first optical filter and the circle center of the second optical filter are located on the same diameter of the rotary bearing.

Optionally, the switching mechanism includes: a carrier; the movable support is movably arranged on the bearing frame, the first optical filter and the second optical filter are positioned on the movable support, and the planes of the first optical filter and the second optical filter are vertical to the light path; the controller is used for driving the movable bearing to linearly move relative to the bearing frame, and the moving direction of the movable bearing is perpendicular to the light path, so that the first light filter and the second light filter are respectively located at the light path position.

Optionally, the controller controls the switching mechanism to periodically adjust the positions of the first optical filter and the second optical filter.

Optionally, the light emitter is a laser, and the laser is a femtosecond laser; the femtosecond laser is used for exciting the femtosecond laser from 700 nanometers to 820 nanometers.

Optionally, the light emitter is a laser, and the laser is a non-tunable picosecond laser; the picosecond laser is used for exciting a picosecond laser with the wavelength of 350 nanometers to 410 nanometers.

Optionally, the first optical filter is a band-pass filter from 420nm to 540nm, and the second optical filter is a long-pass filter from 490nm or a band-pass filter from 500nm to 620 nm.

Compared with the prior art, the embodiment of the utility model provides a technical scheme has following advantage:

in the detecting device provided by the embodiment of the present invention, the positions of the first optical filter and the second optical filter are adjusted by the switching mechanism in the optical filter, so that the first optical filter or the second optical filter is respectively located on the light path, so as to transmit the fluorescence and respectively form a first fluorescence or a second fluorescence, the first fluorescence and the second fluorescence are received by the same photoelectric detector, the controller coordinates the switching mechanism and the photodetector to enable the first optical filter to be positioned on the light path, the photodetector obtains a first fluorescence signal based on the first fluorescence such that when the second optical filter is positioned on the optical path, the photodetector obtains a second fluorescence signal based on the second fluorescence, and the time counter is configured to obtain time-based first fluorescence lifetime data from the first fluorescence signal and is further configured to obtain time-based second fluorescence lifetime data from the second fluorescence signal. With being equipped with dichroic mirror and two photoelectric detector's detection device, dichroic mirror falls into first fluorescence and second fluorescence with fluorescence, and first fluorescence and second fluorescence are compared by the condition that two photoelectric detector received respectively, the embodiment of the utility model provides a, with first light filter and second light filter integration in the light filter, coordinate the position of switching mechanism adjustment first light filter and second light filter on the light path through the controller to let photoelectric detector obtain first fluorescence signal and second fluorescence signal, make a dichroic mirror and a photoelectric detector have been saved at least on the light path, be favorable to simplifying the light path, reduce the cost of manufacture.

In an alternative, the light emitter comprises a laser, and excitation light of the laser is simultaneously located in an absorption spectrum band of NADH and FAD, and can simultaneously enable NADH and FAD to generate fluorescence. The laser device can be adjusted in the detection process by utilizing the detection device, so that the structure of the detection device can be simplified.

In an alternative scheme, the controller controls the switching mechanism to periodically adjust the positions of the first optical filter and the second optical filter, so that fluorescence can periodically penetrate through the first optical filter or the second optical filter to form first fluorescence and second fluorescence respectively, the controller connects the photoelectric detector with the switching mechanism, the first fluorescence signal and the second fluorescence signal can be correspondingly and periodically obtained, the time counter can obtain first fluorescence life data and second fluorescence life data based on time based on the first fluorescence signal and the second fluorescence signal, accuracy and precision of the first fluorescence life data and the second fluorescence life data are high, and detection sensitivity is improved.

Drawings

FIG. 1 is a schematic diagram of fluorescence lifetime;

FIG. 2 is a functional block diagram of a detection device in the disclosed technology;

fig. 3 is a functional block diagram of a detection apparatus according to an embodiment of the present invention;

fig. 4 is a schematic structural diagram of a detection device according to an embodiment of the present invention;

FIG. 5 is a schematic diagram of the first embodiment of the switching mechanism of FIG. 3 showing the structure of the second filter for transmitting fluorescence;

FIG. 6 is a schematic diagram of the first embodiment of the switching mechanism of FIG. 3 showing the structure of the first filter for transmitting fluorescence;

FIG. 7 is a schematic structural diagram of a second embodiment of the switching mechanism of FIG. 3;

FIG. 8 is a schematic structural diagram of a third embodiment of the switching mechanism of FIG. 3;

FIG. 9 is a functional block diagram of the time counter of FIG. 3

FIG. 10 is a result of the detection by the detection apparatus shown in FIG. 4;

fig. 11 is a diagram based on the detection result in the phasor domain obtained by the phasor domain conversion unit.

Detailed Description

As known from the background art, the conventional detection device has a complex optical path and high manufacturing cost. The cause of the above technical problem is now analyzed in conjunction with a detection device of the disclosed technology.

Referring to fig. 2, a schematic structural diagram of a detection device in the disclosed technology is shown.

The detection device comprises: the light-emitting unit 100, the dichroic mirror 104, the first optical filter 101, the second optical filter 102, the first photodetector 111, the second photodetector 112, and the time-dependent single photon counting unit 110. Wherein:

a light emitting unit 100 for providing excitation light capable of generating fluorescence when projected onto the detection object; a dichroic mirror 104 for reflecting light of a first wavelength band in the fluorescence and transmitting light of a second wavelength band in the fluorescence; a first filter 101 for transmitting light of a first wavelength band among the fluorescent light to form first fluorescent light; a second filter 102 for transmitting light of a second wavelength band in the fluorescence to form second fluorescence; a first photodetector 111 for detecting the first fluorescence to form a first fluorescence signal; a second photodetector 112 for detecting the second fluorescence to form a second fluorescence signal; and a time-dependent single photon counting unit 110, configured to obtain time-based first fluorescence lifetime data according to the first fluorescence signal, and further configured to obtain time-based second fluorescence lifetime data according to the second fluorescence signal.

In the detection device, a dichroic mirror 104 is used to reflect light of a first wavelength band in fluorescence, a light region of a second wavelength band is transmitted, so that light of the first wavelength band and light of the second wavelength band are separated, and the light of the first wavelength band and the light of the second wavelength band are respectively transmitted through a first optical filter 101 and a second optical filter 102 to form first fluorescence and second fluorescence, the first fluorescence and the second fluorescence are respectively detected by a first photoelectric detector 111 and a second photoelectric detector 112 to form a first fluorescence signal and a second fluorescence signal, that is, the detection device divides light of the first wavelength band and light of the second wavelength band in fluorescence into two optical paths through the dichroic mirror 104, and the light of the two wavelength bands is matched with one color filter and one photoelectric detector, so that the structure of the detection device is complex, the corresponding detection optical path is complex, and in addition, the detection device is divided into two optical paths according to the light of the first wavelength band and the light of the second wavelength band, the number of optical devices used is large, which results in high manufacturing cost of the detection device.

In order to solve the above technical problem, the detecting device of the embodiment of the present invention adjusts the positions of the first optical filter and the second optical filter by the switching mechanism in the optical filter, so that the first filter or the second filter is respectively positioned on the light path and transmits the fluorescence to respectively form a first fluorescence or a second fluorescence, the first fluorescence and the second fluorescence are received by the same photoelectric detector, and the controller coordinates the switching mechanism and the photoelectric detector to enable the first optical filter to be positioned on the light path, the photodetector obtains a first fluorescence signal based on the first fluorescence such that when the second optical filter is positioned on the optical path, the photodetector obtains a second fluorescence signal based on the second fluorescence, and the time counter is configured to obtain time-based first fluorescence lifetime data from the first fluorescence signal and is further configured to obtain time-based second fluorescence lifetime data from the second fluorescence signal. With being equipped with dichroic mirror and two photoelectric detector's detection device, dichroic mirror falls into first fluorescence and second fluorescence with fluorescence, and first fluorescence and second fluorescence are compared by the condition that two photoelectric detector received respectively, the embodiment of the utility model provides a, with first light filter and second light filter integration in the light filter, coordinate the position of switching mechanism adjustment first light filter and second light filter on the light path through the controller to let photoelectric detector obtain first fluorescence signal and second fluorescence signal, make a dichroic mirror and a photoelectric detector have been saved at least on the light path, be favorable to simplifying the light path, reduce the cost of manufacture.

The technical solutions in the embodiments of the present invention will be described clearly and completely with reference to the accompanying drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only some embodiments of the present invention, not all embodiments.

Referring to fig. 3 and 4, fig. 3 is a functional block diagram of a detecting device according to an embodiment of the present invention, and fig. 4 is a schematic structural diagram of the detecting device according to an embodiment corresponding to fig. 3. The utility model discloses detection device includes:

a light emitter 200 for providing excitation light capable of generating fluorescence when projected onto the detection object; the optical filter 201 comprises a first optical filter 2011, a second optical filter 2012 and a switching mechanism 2013, wherein the switching mechanism 2013 is used for adjusting the positions of the first optical filter 2011 and the second optical filter 2012, so that the first optical filter 2011 or the second optical filter 2012 are positioned on an optical path to transmit fluorescence; the fluorescence passes through the first filter 2011 to form first fluorescence, and the fluorescence passes through the second filter 2012 to form second fluorescence; a photodetector 202 for detecting the first fluorescence or the second fluorescence; the controller 203 is connected to the photodetector 202 and the switching mechanism 2013, and configured to enable the photodetector 202 to detect a first fluorescence when the switching mechanism 2013 controls the first optical filter 2011 to be in the optical path position, so as to obtain a first fluorescence signal; the switching mechanism 2013 is further configured to obtain a second fluorescence signal when the photodetector 202 detects a second fluorescence when the second filter 2012 is controlled to be at the optical path position; a time counter 204 for obtaining time-based first fluorescence lifetime data from the first fluorescence signal and for obtaining time-based second fluorescence lifetime data from the second fluorescence signal.

The positions of the first filter 2011 and the second filter 2012 are adjusted by the switching mechanism 2013 in the filter 201, so that the first filter 2011 or the second filter 2012 are respectively located on the light path, to transmit the fluorescence light to form a first fluorescence light or a second fluorescence light, respectively, which are received by the same photodetector 202, the controller 203 coordinates the switching mechanism 2013 with the photodetector 202, so that when the first filter 2011 is positioned on the light path, the photodetector 202 obtains a first fluorescence signal based on the first fluorescence, so that when the second filter 2012 is located on the light path, the photodetector 202 obtains a second fluorescence signal based on the second fluorescence, a time counter 204, for obtaining time-based first fluorescence lifetime data from the first fluorescence signal and for obtaining time-based second fluorescence lifetime data from the second fluorescence signal. With being equipped with dichroic mirror and two photoelectric detector's detection device, dichroic mirror falls into first fluorescence and second fluorescence with fluorescence, and first fluorescence and second fluorescence are compared by the condition that two photoelectric detector received respectively, the embodiment of the utility model provides a, with first light filter 2011 and second light filter 2012 integration in light filter 201, coordinate switching mechanism adjustment first light filter 2011 and second light filter 2012 on the light path position through controller 203, and let photoelectric detector 202 obtain first fluorescence signal and second fluorescence signal, make a dichroic mirror and a photoelectric detector 202 have been saved at least on the light path, be favorable to simplifying the light path, reduce the cost of manufacture.

It should be noted that, in this embodiment, two coenzymes are detected in the cervical tissue section (i.e. the detection object is the cervical tissue section): fluorescence of Nicotinamide Adenine Dinucleotide (NADH) and Flavin Adenine Dinucleotide (FAD) will be exemplified. In other embodiments, fluorescence corresponding to other molecules may also be detected.

In this embodiment, the light emitter 200 is a laser. The exciting light provided by the laser is simultaneously positioned in NADH and FAD absorption spectrum bands, so that NADH and FAD can generate fluorescence simultaneously. The fluorescence generated after the excitation light provided by the laser is projected onto the detection object is respectively formed into the first fluorescence and the second fluorescence after passing through the first optical filter 2011 and the second optical filter 2012, so that the laser does not need to be adjusted in the detection process by using the detection device, and the structure and the detection method of the detection device can be simplified.

In this embodiment, the light emitter 200 is a picosecond laser, and is configured to emit a picosecond laser (for example, a 400nm laser) with a wavelength of 350nm to 410nm, where the picosecond laser is configured to excite NADH in a cervical tissue slice to reach an excited state, and then emit first fluorescence, and is configured to excite FAD in the cervical tissue slice to reach the excited state, and then emit second fluorescence. In other embodiments, the light emitter is a non-tunable femtosecond laser for emitting a femtosecond laser of 700nm to 820nm, the femtosecond laser is used for exciting NADH in the cervical tissue slice to an excited state to emit a first fluorescence, and is used for exciting FAD in the cervical tissue slice to an excited state to emit a second fluorescence.

In this embodiment, the filter 201 is used to transmit the first fluorescence or the second fluorescence. Compared with the detection device provided with a dichroic mirror and two photodetectors, in the detection device, the dichroic mirror divides fluorescence into first fluorescence and second fluorescence, and the first fluorescence and the second fluorescence are received by the two photodetectors respectively, the optical filter 201 of the present embodiment integrates the first optical filter 2011 and the second optical filter 2012 together, and the switching mechanism 2013 adjusts the position of the first optical filter 2011 or the second optical filter 2012, so that the fluorescence passing through the optical filter 201 can periodically provide the first fluorescence or the second fluorescence to the photodetector 202, which is beneficial to simplifying a light path and reducing the manufacturing cost.

Specifically, the first filter 2011 of the filter 201 is configured to transmit light of a first wavelength band of the fluorescent light to form first fluorescent light. In this embodiment, for NADH, the first filter 2011 may be a band pass filter of 420nm to 540nm, because the first filter 2011 may pass light within an NADH emission spectrum band.

Specifically, the second filter 2012 in the filter 201 is configured to transmit light of a second wavelength band in the fluorescent light to form second fluorescent light. In this embodiment, for the FAD, the second filter 2012 only needs to enable light in an emission spectrum band of the FAD to pass through, and therefore, the second filter 2012 may be a long-pass filter greater than 490nm or a band-pass filter from 500nm to 620 nm.

In other embodiments, for other species of detection molecules, a filter corresponding to the emission band of the molecule may be selected for passing the fluorescence generated by the excited molecule.

In this embodiment, the switching mechanism 2013 includes: a carriage (not shown); a rotating support (not shown in the figures) rotatably disposed on the carrier, wherein the first optical filter 2011 and the second optical filter 2012 are located on the rotating support, and planes of the first optical filter 2011 and the second optical filter 2012 are both perpendicular to the optical path; and the controller 203 is used for driving the rotary bearing to rotate relative to the bearing frame, and the rotary plane of the rotary bearing is perpendicular to the light path, so that the first light filter 2011 and the second light filter 2012 are respectively positioned at the light path positions.

When the detection device works, the controller 203 drives the rotation support to rotate relatively to the bearing frame, and accordingly the positions of the first optical filter 2011, the second optical filter 2012 and the light path on the rotation support are changed, so that the first optical filter 2011 and the second optical filter 2012 are respectively located on the light path, and the fluorescence can be respectively transmitted through the first optical filter 2011 or the second optical filter 2012.

It should be noted that the plane where the first optical filter 2011 and the second optical filter 2012 are located is perpendicular to the optical path, and what is indicated by the perpendicular direction of the rotation plane of the rotation bearing and the optical path is that, when the detection device works, the fluorescence penetrates through the first optical filter 2011 and the second optical filter 2012, so that the accuracy and precision of the subsequently obtained first fluorescence life data and second fluorescence life data can be improved, and the detection sensitivity is improved.

As shown in fig. 5 and fig. 6, which illustrate a schematic structure of a first embodiment of the switching mechanism in the present embodiment, specifically, fig. 5 is a schematic structure of the first embodiment of the switching mechanism in fig. 3, in which fluorescence passes through the second filter 2012; fig. 6 is a schematic diagram illustrating a structure of a first embodiment of the switching mechanism in fig. 3 for transmitting fluorescence through a first filter 2011.

In a first embodiment, the rotating bearing is a circular ring, the center of rotation of the rotating bearing is the center of the circle of the rotating bearing, a straight line passing through the center of the circle of the rotating bearing divides the rotating bearing into a first region and a second region, the first optical filter 2011 is located in the first region, and the second optical filter 2012 is located in the second region.

In this embodiment, the first region and the second region are divided into semi-circles, that is, the first region and the second region have equal areas, and correspondingly, the first filter 2011 and the second filter 2012 have equal areas. First region and second region make full use of the region of rotation bearing makes the area of first light filter 2011 and second light filter 2012 is great, and in detection device working process, fluorescence passes through first light filter 2011 or second light filter 2012 all the time, thereby photodetector can detect first fluorescence or second fluorescence always, photodetector 202 obtains not having the neutral gear period between first fluorescence signal and the second fluorescence signal, that is to say photodetector 202 can obtain continuous fluorescence signal, thereby time counter 204 can obtain more first fluorescence life data and second fluorescence life data, is favorable to making the accuracy and the precision of first fluorescence life data and second fluorescence life data higher, has improved and has detected the sensitivity.

In this embodiment, the distance from the position 300 where the fluorescence passes through the first filter 2011 or the second filter 2012 to the center of the circle is one third to two thirds of the radius of the circle. If the ratio is too large, the position where the fluorescence penetrates through the first optical filter 2011 or the second optical filter 2012 is close to the edge of the rotating support, and if a slight deviation exists in the light path of the detection device, the fluorescence cannot penetrate through the first optical filter 2011 or the second optical filter 2012, so that the photoelectric detector 202 cannot obtain the first fluorescence signal and the second fluorescence signal, and the corresponding time counter 204 cannot obtain the first fluorescence lifetime data and the second fluorescence lifetime data. If the proportion is too small, the position of the fluorescence passing through the first optical filter 2011 or the second optical filter 2012 is too close to the center of the rotating bearing, if a slight deviation exists in the light path of the detection device, the situation that the preset fluorescence passes through the first optical filter 2011 but passes through the second optical filter 2012 occurs, or the situation that the fluorescence passes through the first optical filter 2011 or the second optical filter 2012 all the time occurs, so that the photoelectric detector 202 cannot accurately obtain the first fluorescence signal and the second fluorescence signal, the accuracy and precision of the first fluorescence life data and the second fluorescence life data obtained by the corresponding time counter 204 are poor, and the detection sensitivity is low.

In this embodiment, when the switching mechanism 2013 works, the rotation support rotates clockwise relative to the carrier, and the corresponding first optical filter 2011 and the second optical filter 2012 rotate clockwise. In other embodiments, the pivoting support may also pivot counterclockwise relative to the carriage.

As shown in fig. 7, which illustrates a schematic structural diagram of a second embodiment of the switching mechanism in this embodiment, the first optical filter 3011 and the second optical filter 3012 are both one, the first optical filter 3011 and the second optical filter 3012 are spaced apart, and the first optical filter 3011 and the second optical filter 3012 are both circular.

The circular first optical filter 3011 and the circular second optical filter 3012 are common type optical filters, and there is no need to customize the optical filters, so that the manufacturing cost of the optical filters can be reduced.

In this embodiment, when the switching mechanism works, the rotation support rotates clockwise relative to the supporting frame, and the corresponding first optical filter 3011 and second optical filter 3012 rotate clockwise. In other embodiments, the pivoting support may also pivot counterclockwise relative to the carriage.

In this embodiment, the frame of the rotary support is circular, the rotation center of the rotary support is the center of the circle of the rotary support, the first optical filter 3011 and the second optical filter 3012 are located on both sides of the center of the rotary support, and the center of the circle of the first optical filter 3011 and the center of the circle of the second optical filter 3012 are located on the same diameter of the rotary support. The first optical filter 3011 and the second optical filter 3012 are arranged on the rotary support in such a way, when the detection device works, the time interval between the movement of the position of the fluorescence on the rotary support from the first optical filter 3011 to the movement of the fluorescence into the second optical filter 3012 is the same as the time interval between the movement of the position of the fluorescence on the rotary support from the second optical filter 3012 to the movement of the fluorescence into the first optical filter 3011, which is beneficial to making the time counter obtain the first fluorescence life data of the first fluorescence signal and the second fluorescence life data of the second fluorescence signal according to a certain period, so that the controller can conveniently and accurately control the photoelectric detection opportunity, thereby further improving the detection accuracy and precision and further improving the detection sensitivity.

The photoelectric detector is used for detecting the first fluorescence which passes through the first optical filter 3011, so that an optical signal is converted into an electric signal to form a first fluorescence signal; and is further configured to detect the second fluorescence passing through the second optical filter 3012, so that the optical signal is converted into an electrical signal to form a second fluorescence signal.

Specifically, the photodetectors are each a photomultiplier tube (PMT), and can convert relatively weak light into an electrical signal with a relatively large gain, thereby improving detection sensitivity.

In other embodiments, the photodetector may also be other photodetectors such as a photodiode, a Charge Coupled Device (CCD), an Avalanche Photodiode (APD), and the like.

The double-channel PMT is arranged in the embodiment, so that the change conditions of two coenzyme factors, namely NADH and FAD, can be obtained simultaneously, more detail information of early tumor lesions is provided, and the missed diagnosis rate caused by small lesion regions is greatly reduced.

As shown in fig. 8, a schematic structural view of a third embodiment of the switching mechanism is shown.

The switching mechanism includes: a carrier; the movable support is movably arranged on the bearing frame, the first optical filter 4011 and the second optical filter 4012 are positioned on the movable support, and the planes of the first optical filter 4011 and the second optical filter 4012 are vertical to the light path; the controller is used for driving the movable bearing to linearly move relative to the bearing frame, the plane where the movable bearing is located is perpendicular to the light path, and the moving direction of the movable bearing is perpendicular to the light path, so that the first light filter and the second light filter are located on the light path respectively.

When the detection device works, the controller drives the moving support to move relatively to the bearing frame in a reciprocating mode, and accordingly the positions of the first light filter 4011 and the second light filter 4012 on the moving support and the positions of the light paths are changed, so that the fluorescence can penetrate through the first light filter 4011 or the second light filter 4012, and the photoelectric detector 202 can detect the first fluorescence and the second fluorescence respectively.

It should be noted that the plane where the first optical filter 4011 and the second optical filter 4012 are located is perpendicular to the optical path, and the moving direction of the moving support is perpendicular to the optical path, that is, when the detection device works, the fluorescence penetrates through the first optical filter 4011 and the second optical filter 4012, so that the energy loss of the fluorescence after passing through the first optical filter 4011 and the second optical filter 4012 is reduced, the accuracy and precision of the subsequently obtained first fluorescence life data and second fluorescence life data can be improved, and the detection sensitivity is improved.

The controller 203 is connected to the photodetector 202 and the switching mechanism 2013.

The controller 203 controls the switching mechanism to periodically adjust the positions of the first optical filter 2011 and the second optical filter 2012, so that the fluorescent light can periodically penetrate through the first optical filter 2011 or the second optical filter 2012 to form first fluorescent light and second fluorescent light respectively, the controller 203 connects the photodetector 202 with the switching mechanism, the first fluorescent signal and the second fluorescent signal can be periodically obtained correspondingly, the time counter 204 can obtain first fluorescent life data and second fluorescent life data based on time based on the first fluorescent signal and the second fluorescent signal, the accuracy and precision of the first fluorescent life data and the second fluorescent life data are high, and the detection sensitivity is improved.

The controller 203 includes a central processing unit, and a memory, a control circuit and a power control hardware board card connected to the central processing unit, the power control hardware board card is connected to the photodetector 202.

The power control hardware board card is used for limiting the line overload power consumption; the central processing unit is an 16/32-bit singlechip processor based on an ARM core; the memory stores control parameters for controlling the switching mechanism and the periodic adjustment of the photoelectric detector 202, and the control parameters in the memory can be called by the central processing unit in real time through the control circuit in the detection process.

In the first and second embodiments, the controller is further connected to a motor, which is a part of the rotary support and is used for driving the first color filter and the second color filter to rotate.

In a third embodiment, the controller is connected to a microplate oscillator, and the microplate oscillator is a part of the movable support and is used for driving the first color filter and the second color filter to reciprocate.

Referring collectively to fig. 9, fig. 9 shows a functional block diagram of the time counter 204, the time counter 204 including a processor 2041 and a driver 2042.

The time counter 204 processes the fluorescence signals (the electrical signal corresponding to the first fluorescence and the electrical signal corresponding to the second fluorescence) collected by the photodetector 202, so as to obtain a first fluorescence lifetime curve and a second fluorescence lifetime curve, so as to further determine whether the fluorescence lifetime of two coenzyme factors, namely NADH and FAD, changes abnormally, and to determine whether a cervical tissue section is diseased.

The processor 2041 is configured to perform data processing on the signals detected and collected by the photodetector 202.

Specifically, the processor 2041 is a time-dependent single photon counting card installed in a computer.

It should be noted that the fluorescence lifetime can be described by a single e-index formula,

I(t)＝I₀exp(-t/τ)

where τ is the fluorescence lifetime, I₀The maximum fluorescence intensity value emitted, i (t), is the fluorescence intensity at time t.

Based on the fitting results, the weighted average lifetime of each pixel in the image acquired by the photodetector 202 is obtained by the following formula:

wherein t is_iFluorescence lifetime of different composition, a_iIs the ratio of different components.

In this embodiment, the driver 2042 is further connected to the scanning unit 206 (shown in fig. 4) and the processor 2041, and is configured to synchronize the scanning of the scanning unit 206 with the processor 2041.

Specifically, the driver 2042 is an external board card connected to the computer through a USB.

It should be noted that, as shown in fig. 4, a first conjugate aperture 209 is further disposed on the optical path between the light emitter 200 and the detection object for filtering the fluorescence of the non-focal plane, so as to improve the detection accuracy and resolution. In other embodiments, the first conjugate orifice may not be provided.

In this embodiment, a dichroic mirror 205 is further disposed on an optical path between the light emitter 200 and the cervical tissue slice, and is used for distinguishing the excitation light from the fluorescence.

Specifically, the dichroic mirror 205 includes a first surface and a second surface opposite to the first surface, the laser light emitted by the laser is incident on the first surface and is transmitted, and after reaching the cervical tissue slice, first fluorescence and second fluorescence are formed, and the first fluorescence and the second fluorescence reach the second surface and are reflected, so that the discrimination between the excitation light and the fluorescence is realized.

In this embodiment, by providing the dichroic mirror 205, interference of the excitation light on the fluorescence can be reduced, thereby improving the detection accuracy. In other embodiments, the detection device may not be provided with the dichroic mirror, so as to simplify the optical path.

With continuing reference to fig. 4, the detecting apparatus of the present embodiment further includes: a scanning unit 206, configured to scan the cervical tissue slice with excitation light, or control the excitation light to detect a preset position of the cervical tissue slice.

Specifically, the scanning unit 206 is a galvanometer driving system, which includes an x-direction galvanometer and a y-direction galvanometer, and is configured to perform surface scanning on the cervical tissue slice.

The scanning unit 206 can control the excitation light to scan the preset position of the cervical tissue slice in addition to the surface scanning, so that the excitation light can detect the specific position of the detected object.

In this embodiment, an objective lens group 207 is further disposed between the scanning unit 206 and the object to be detected, and is used for magnifying the image of the cervical tissue slice, so that a magnified image of the cervical tissue slice can be obtained, and thus, the detection position of the cervical tissue slice is determined, and the detection accuracy is further improved.

The objective lens group 207 is further configured to transmit the fluorescence, so that the fluorescence is transmitted to the photodetector 202 of the subsequent optical path for detection.

Specifically, the objective lens group 207 is an objective lens of a confocal laser microscope.

It should be noted that, in other embodiments, by setting the self-focusing motor and the objective lens to adjust the focal helix, continuous scanning acquisition in the z direction of the cervical tissue slice can be realized, so that one more dimension of the cervical tissue slice can be detected, and similar detection accuracy can be obtained for tissues at different depths.

As shown in fig. 4, the examination apparatus further comprises a stage 208 for placing the examination object cervical tissue slice.

In other embodiments, the detection device may not be provided with the stage, but may include an optical fiber for transmitting the excitation light to the detection object and for transmitting the fluorescence. Thereby realizing the detection of the detected object through the optical fiber. The optical fiber is made of soft materials, and can detect organs which are not beneficial to detection through a probe at the end part of the optical fiber.

When the exciting light reaches the cervical tissue section, molecules (including NADH, FAD, collagen, elastin, retinol and the like) in the cervical tissue section are excited to an excited state, then the cervical tissue section returns to a ground state in a radiation transition mode, fluorescence of the NADH and FAD is filtered through an optical filter, so that fluorescence life information of the NADH and FAD is measured, and whether the cervical tissue is diseased or not is judged by judging the change of the fluorescence life of the NADH and FAD.

The principle of judging whether cervical tissues are diseased or not through the change of the fluorescence life of NADH and FAD is as follows: the cells in cervical tissue section require energy to maintain homeostasis, all processes of cellular activity are energy dependent, the main energy producing activities are glycolysis and oxidative phosphorylation, and two coenzyme factors, NADH and FAD, are electron donors and acceptors involved therein. NADH and FAD have two states in the organism, one is in a bound state (bound) and the other is in a free radical state (free). The life of NADH and FAD in free radical state is hundreds of picoseconds, and the life of FAD and NADH in free radical state is thousands of picoseconds. The energy metabolism activity of normal cells differs significantly from the cellular metabolism in pathological conditions. Tumor cells are characterized by hyperactive and uncontrollable cell proliferation. When the metabolism of the microenvironment where the cells are located is abnormal, the autofluorescence lifetime of NADH and FAD is changed. Therefore, whether metabolism in a human body is abnormal can be judged by detecting the ratio of the combination states of the two coenzymes and the change of the service life, and further, whether organs are diseased or not can be judged.

In order to measure the fluorescence lifetime, in this embodiment, the excitation light causes the cervical tissue slice to generate fluorescence, the fluorescence reaches the dichroic mirror 205 through the objective lens group 207 and the scanning unit 206, and the dichroic mirror 205 reflects the fluorescence, so that the fluorescence is distinguished from the excitation light on the optical path, and the subsequent optical path detects the fluorescence.

It should be noted that, in the detection apparatus of this embodiment, the fluorescence is reflected by the dichroic mirror 205 and then directly permeates through the optical filter 201 to reach the photodetector 202, and no reflecting mirror is arranged between the dichroic mirror 205 and the photodetector 202, which is beneficial to reducing the use of optical devices, and can reduce the cost of the detection apparatus and simplify the optical path. In other embodiments, a reflecting mirror may be further disposed between the dichroic mirror and the photodetector on the optical path, and the reflecting mirror is used to change the direction of light propagation, so as to improve the compactness of the detection apparatus.

With continuing reference to fig. 4, in this embodiment, the detecting device further includes: an electrical signal coupling unit 214 for transmitting the electrical signal detected by the photodetector 202 to the time counter 204.

In other embodiments, the electrical signal coupling unit may not be provided.

When malignant tumors occur in organs, the adjacent regions of the diseased organ are also affected. Taking ovary and uterus as examples, when there is malignant tumor in ovary or uterus, metabolism of cervix shows abnormality, further influencing microenvironment of cervix tissue. Similarly, when the digestive organs are diseased, the metabolism of the oral tissues is also altered.

With continued reference to fig. 4, the detection apparatus may further include: a data processing unit 221, configured to process the first fluorescence lifetime data and the second fluorescence lifetime data, and determine whether a processing result falls within a reference range, where the reference range is obtained based on data corresponding to a distal portion, where the distal portion is a portion that is harder to detect than the detection object, or the reference range is obtained based on data corresponding to the detection object.

In this embodiment, the data processing unit 221 is disposed in a computer.

It should be noted that, in the case where the reference range is obtained based on data corresponding to the distal site, the reference range is obtained from data of a healthy normal person and a patient with a lesion at the distal site. For example, the distal site refers to the ovary, the test object is a section of cervical tissue, and the ovary is a relatively difficult site to detect relative to the cervical tissue. The reference range is set by the data of healthy normal people and patients with ovarian diseases (the data can be the detection data obtained by the detection device of the utility model in advance, and also can be the reference data obtained according to the literature). When the detection is carried out, the cervical tissue section is used as a detection object, fluorescence lifetime data of coenzyme or other molecules is obtained by detecting the cervical tissue section, and whether the ovary is abnormal is judged based on whether the fluorescence lifetime data falls into the reference range.

The detection result of the far-end part (such as ovary or digestive tract organ) can be obtained by detecting the detection object (such as cervical tissue or oral tissue), the detection difficulty of the detection object is relatively low, and especially for the detection of the far-end position with difficult biopsy positions, the embodiment of the utility model can reduce the detection difficulty and even avoid traumatic biopsy operation.

On the other hand, the reference range may be obtained based on data corresponding to the detection object, and refers to a reference range obtained from data of a normal person who is healthy and a patient in which a lesion occurs in the detection object. For example, the test object is a cervical tissue section. The reference range is set by data of a healthy normal person and a patient with a lesion of cervical tissue. When the detection is carried out, the cervical tissue is used as a detection object, fluorescence lifetime data of coenzyme or other molecules is obtained by detecting the cervical tissue, and whether the cervical tissue is abnormal or not is judged based on whether the fluorescence lifetime data falls into the reference range or not.

It should be noted that the reference range includes one or more numerical ranges, and in the case where the reference range is a plurality of numerical ranges, by determining which numerical range the detected data falls into, the abnormal situation can be more finely distinguished.

Specifically, in order to combine the obtained first fluorescence lifetime data and the obtained second fluorescence lifetime for analysis, the data processing unit 221 may establish a coordinate system having the first fluorescence lifetime as an abscissa (ordinate) and the second fluorescence lifetime as an ordinate (abscissa), set the measured first fluorescence lifetime data and the second fluorescence lifetime data into the coordinate system as detection data points, and also set the data of the distal end portion as reference data into the coordinate system as reference data points, may obtain a reference range based on the distribution of the reference points, and may obtain a determination result by determining whether the detection data points fall within the reference range.

Referring to fig. 10, the result of the detection by the detection apparatus shown in fig. 4 is shown. Wherein the abscissa is the lifetime of the bound FAD autofluorescence, and the ordinate is the lifetime of the bound NADH autofluorescence. The square data points are fluorescence lifetime data of the binding state FAD and the binding state NADH of the normal human cervix (reference data points obtained by the detection device), and the circular data points (detection results obtained by the detection device) are fluorescence lifetime data of the binding state NADH (first fluorescence lifetime data) and fluorescence lifetime data of the binding state FAD (second fluorescence lifetime data) obtained from the cervix tissue of the patient with ovarian cancer. The fluorescence lifetime data of FAD and NADH detected include fluorescence lifetime data of the bound state. The values for the individual data points in FIG. 10 are fluorescence lifetime data for the bound state.

The results of fig. 10 show that the fluorescence lifetime data of NADH (first fluorescence lifetime data) and the fluorescence lifetime data of FAD (second fluorescence lifetime data) obtained by detecting cervical tissue have a certain difference in the value distribution from the reference data, and the reference range can be set by using the dotted line in the figure as a boundary. During detection, if the detected data point falls into the lower left of the dotted line, the data point is abnormal, and the data point falls into the upper right of the dotted line, the data point is normal. This demonstrates that detection of the test substance (cervical tissue) can determine whether or not an abnormality has occurred in a distal organ (ovary) that is difficult to detect.

Because the detection accuracy of the detection device of the embodiment is higher, the detection of the ovarian lesion can be realized by detecting the cervical tissue. Cervical tissue can be detected by colposcopy, which reduces the difficulty of detection compared to obtaining tissue from the uterus or from the distal organs, such as the ovary.

Similarly, when other organs are detected, the organs far away from the malignant tumor can be detected more easily to find the tumor deep in the abdominal cavity. For example: by detecting the oral cavity, detection data are provided for judging esophageal lesions.

In actual operation, the detection object may be detected, and the reference range may be obtained based on data corresponding to the detection object, so as to determine whether the detection object itself is abnormal.

The phasor spectrometry can eliminate the influence of impurities such as blood in a complex environment on data, performs real-time data self-detection and correction, has stronger adaptability and specificity, and helps to improve the diagnosis sensitivity of early lesions or precancerous lesions.

With continued reference to fig. 3 and 4, to further improve detection accuracy. The detection apparatus may further include a phasor domain conversion unit 220 connected to the time counter 204, for converting the time domain data of the time counter 204 into a phasor spectrum for analysis.

The specific fitting formula is as follows:

here, i, j is defined as one pixel of the image, ω is the angular frequency, ω is 2 pi f, and f is the repetition frequency of the laser.

The position of the phasor curve is directly linked with the lifetime, and the fluorescence lifetime obtained through the single e index in the time counter 204 can find the corresponding position on a semicircle with the center coordinates of (0.5, 0) and the radius of 0.5 after phasor conversion.

Specifically, the phasor domain conversion unit 220 is configured to convert the first fluorescence lifetime data and the second fluorescence lifetime data from a time domain to a phasor spectrum, so as to obtain first phasor spectrum information corresponding to the first fluorescence and second phasor spectrum information corresponding to the second fluorescence.

By converting the fluorescence lifetime data from the time domain to the phasor domain, the measured fluorescence lifetime curve can be converted into a phasor, the phase being the angle of the phasor, and the modulation being the amplitude of the phasor. Through analyzing the detected data in the phasor domain, the influence on blood, lymph fluid and the like in the organ detection process can be eliminated, so that the detection accuracy and precision are further improved.

Each image obtained by the time counter 204 is converted into a scatter diagram in a phasor coordinate system by the phasor domain conversion unit 220. For each acquisition area, the density distribution of a scatter diagram in a phasor coordinate system can be obtained, and the position with the most concentrated density distribution is selected as the central coordinate value of the acquisition area.

After the obtained fluorescence lifetime data is converted into the phasor spectrum by the phasor domain conversion unit 220, the data processing unit may determine whether the detected object is abnormal based on the phasor spectrum information corresponding to the first fluorescence lifetime data and the second fluorescence lifetime data.

Referring to fig. 11, a result of processing by the data processing unit 221 based on information obtained by the phasor domain conversion unit 220 is shown. The difference from fig. 10 is that the reference range in fig. 11 is a reference range obtained based on data corresponding to the detection object. Specifically, the object to be detected is a specimen (cervical tissue slice) corresponding to an organ which is easy to detect, and the reference range is set by data acquired from the organ which is easy to detect.

Normal represents the test result of a Normal person (the test result obtained by the detection device), and CIN3 represents the test result obtained by the device of the embodiment of the present invention for a patient with high risk of cervical cancer (high risk precancerous lesion), and the fluorescence lifetime data obtained by the two before the phasor domain conversion is performed are relatively close. In the phasor domain coordinate system shown in fig. 11, CIN3 is located at the lower right of the Normal value, so that the detection data of the high risk group of cervical cancer before cancer is distinguished from the detection data of the Normal person, and effective diagnosis of the high risk group of cervical cancer before lesion is conveniently realized. Here, CIN3 is a result of a phasor domain analysis including information on the fluorescence lifetime of the first fluorescence and the fluorescence lifetime of the second fluorescence, and specifically, is a result obtained by combining the fluorescence lifetime data of the first fluorescence and the fluorescence lifetime data of the second fluorescence.

Fig. 11 schematically shows a reference range obtained by detecting a detection object and based on data corresponding to the detection object, and further, whether or not abnormality is determined. In other embodiments, the detection object may be detected, and the reference range is a reference range of the phasor spectrum obtained through data corresponding to the distal portion, so as to determine whether the distal portion is abnormal through the data detected from the detection object.

It should be noted that in fig. 10 and 11, the reference range is one, and in other embodiments, the reference range is a plurality of ranges, for example, different reference ranges are ranges set for data of different lesion stages, and the relationship between the processing result obtained by the detection and the reference range is used to realize more detailed detection.

Although the embodiments of the present invention are disclosed above, the embodiments of the present invention are not limited thereto. Various changes and modifications may be effected therein by one of ordinary skill in the pertinent art without departing from the scope or spirit of the embodiments of the present invention, and it is intended that the scope of the embodiments of the present invention be defined by the appended claims.

Claims

1. A detecting device for detecting a detection object, comprising:

a light emitter for providing excitation light, wherein the excitation light projected on the detection object can generate fluorescence;

the optical filter comprises a first optical filter, a second optical filter and a switching mechanism, wherein the switching mechanism is used for adjusting the positions of the first optical filter and the second optical filter so that the first optical filter or the second optical filter is positioned on the light path to transmit fluorescence; the fluorescence penetrates through the first optical filter to form first fluorescence, and the fluorescence penetrates through the second optical filter to form second fluorescence;

a photodetector for detecting the first fluorescence or the second fluorescence;

the controller is connected with the photoelectric detector and the switching mechanism and used for enabling the photoelectric detector to detect first fluorescence to obtain a first fluorescence signal when the switching mechanism controls the first optical filter to be in the position of the optical path; the switching mechanism is further used for controlling the second optical filter to be in the optical path position, so that when the photoelectric detector detects second fluorescence, a second fluorescence signal is obtained;

and the time counter is used for obtaining first fluorescence lifetime data based on time according to the first fluorescence signal and is also used for obtaining second fluorescence lifetime data based on time according to the second fluorescence signal.

2. The detection device of claim 1, wherein the switching mechanism comprises:

a carrier;

the rotating support is rotatably arranged on the bearing frame, the first optical filter and the second optical filter are positioned on the rotating support, and the planes of the first optical filter and the second optical filter are both vertical to the light path;

the controller is used for driving the rotary bearing to rotate relative to the bearing frame, and a rotating plane of the rotary bearing is perpendicular to the light path, so that the first light filter and the second light filter are respectively located at the light path position.

3. The detecting device according to claim 2, wherein the rotating support is a circular ring, the center of rotation of the rotating support is the center of the circle of the rotating support, a straight line passing through the center of the circle of the rotating support divides the rotating support into a first region and a second region, the first optical filter is located in the first region, and the second optical filter is located in the second region.

4. The detecting device for detecting the rotation of a motor rotor as claimed in claim 3, wherein the distance from the position where the fluorescence passes through the first filter or the second filter to the center of the circle is one third to two thirds of the radius of the circle.

5. The detecting device for detecting the rotation of a motor rotor as claimed in claim 2, wherein the first optical filter and the second optical filter are both one and spaced apart from each other, and the first optical filter and the second optical filter are both circular.

6. The detecting device according to claim 5, wherein the frame of the rotating holder is a circle, the center of rotation of the rotating holder is a center of the circle of the rotating holder, the first optical filter and the second optical filter are located at two sides of the center of the circle of the rotating holder, and the center of the circle of the first optical filter and the center of the circle of the second optical filter are located on the same diameter of the rotating holder.

7. The detection device of claim 1, wherein the switching mechanism comprises:

a carrier;

the movable support is movably arranged on the bearing frame, the first optical filter and the second optical filter are positioned on the movable support, and the planes of the first optical filter and the second optical filter are vertical to the light path;

the controller is used for driving the movable bearing to linearly move relative to the bearing frame, and the moving direction of the movable bearing is perpendicular to the light path, so that the first light filter and the second light filter are respectively located at the light path position.

8. The detecting device for detecting the rotation of a motor rotor as claimed in claim 1, wherein the controller controls the switching mechanism to periodically adjust the positions of the first filter and the second filter.

9. The detection device according to any one of claims 1 to 8, wherein the light emitter is a laser, and the laser is a femtosecond laser;

the femtosecond laser is used for exciting the femtosecond laser from 700 nanometers to 820 nanometers.

10. The device according to any one of claims 1 to 8, wherein the light emitter is a laser, the laser being a non-tunable picosecond laser;

the picosecond laser is used for exciting a picosecond laser with the wavelength of 350 nanometers to 410 nanometers.

11. The detecting device for detecting the rotation of a motor rotor as claimed in claims 1 to 8, wherein the first filter is a band-pass filter of 420nm to 540nm, and the second filter is a long-pass filter of 490nm or a band-pass filter of 500nm to 620 nm.