CN205280574U - Multi -functional optoacoustic, fluorescence is micro - and fluorescence spectra formation of image analytical equipment - Google Patents

Abstract

The utility model provides a multifunctional photoacoustic, fluorescence microscope and fluorescence spectrum imaging analysis device. The utility model controls the laser scanning device to scan point by point or the three-dimensional precision electric translation platform to scan point by point, and the laser pulse energy is deposited in the sample. The thermal expansion of the sample after absorbing the laser pulse energy produces an ultrasonic signal. The ultrasonic signal is converted into an electrical signal by the ultrasonic transducer and amplified by the signal amplifier. The data acquisition system collects and records the ultrasonic signal emitted by the sample at each scanning point. At the same time, the CCD camera in the spectrometer synchronously collects the spectral information of the back-facing fluorescence of the sample at each scanning point, and the data acquisition system records the spectral information of the spectrometer at each scanning point to complete the sample plane (<i>xy</i> plane ) After the data collection of the last area to be tested, the computer program will generate photoacoustic microscopic images, fluorescent microscopic images, and fluorescent microspectral images of the sample.

Description

Multifunctional Photoacoustic, Fluorescence Microscopy and Fluorescence Spectrum Imaging Analysis Device

technical field

The utility model relates to a multifunctional photoacoustic, fluorescent microscope and fluorescent spectrum imaging analysis device, which belongs to the technical field of microscopic imaging.

Background technique

Microscopic imaging technology is often used to observe samples that cannot be observed by the naked eye. The application fields include biomedicine, chemistry, physics, metallurgy, measurement and other fields, and it has a unique position in the development of many fields. In the process of continuous development and improvement of microscope technology, from the initial field microscope to the later confocal microscope, major breakthroughs have been made in terms of magnification and anti-noise. However, whether it is a traditional field microscope or a confocal microscope, the light intensity of the sample's scattered light is used to image the sample, and the traditional microscopic imaging can only obtain the structural image of the sample. Even fluorescence imaging can only obtain the microscopic image of the fluorescent light intensity of the sample, and cannot obtain the spectral characteristic information of the sample; the absorption characteristic information of the sample to light cannot be obtained. Photoacoustic imaging technology is an imaging technology developed by using the photoacoustic effect of ultrasonic signals generated by the deposition of light in biological tissues and absorbing media. This imaging technology can achieve optical resolution, and its resolution is much higher than that of ultrasonic imaging technology. Since the photoacoustic effect is closely related to the absorption characteristics of the sample, the characteristic information of the light absorption of the sample can be obtained, which has potential application value in functional imaging technology.

Contents of the invention

The purpose of this utility model is to overcome the deficiencies in the prior art, to provide a multi-functional photoacoustic, fluorescence microscope and fluorescence spectrum imaging analysis device, which can simultaneously realize photoacoustic microscope imaging, fluorescence microscope imaging and Fluorescence microscopy imaging.

According to the technical solution provided by the utility model, the multifunctional photoacoustic, fluorescent microscope and fluorescent spectrum imaging analysis device is characterized in that it includes a three-dimensional electric precision translation stage capable of moving in three directions of x, y and z , laser scanning device, pulsed laser light source, spectrometer, ultrasonic signal acquisition system and spectral signal acquisition system; a water box for placing samples is set on the three-dimensional electric precision translation stage, and an ultrasonic signal window is arranged at the bottom of the water box , the first ultrasonic transducer and the second ultrasonic transducer are respectively arranged at the bottom and side of the ultrasonic signal window, the first ultrasonic transducer and the second ultrasonic transducer are respectively connected to the signal amplifier through the signal switching switch, and the signal amplifier is connected to Ultrasonic signal data acquisition system;

A laser scanning device capable of moving the laser in the x and y directions is arranged above the three-dimensional electric precision translation stage, and an objective lens is arranged between the laser scanning device and the three-dimensional electric precision translation stage; A thermal mirror, a beam splitter mirror and a pulsed laser light source are arranged in sequence on one side; the spectral signal data acquisition system is connected to the CCD camera of the spectrometer.

Further, the size of the ultrasonic signal window is larger than the size of the sample.

Further, the ultrasonic signal window is covered with a thin film that can transmit ultrasonic waves.

Further, the ultrasonic signal data acquisition system and the spectral signal data acquisition system are respectively connected to the electrical signal output ends of the photodetectors.

Further, the three-dimensional electric precision translation stage, laser scanning device, pulsed laser light source, ultrasonic signal data acquisition system and spectrum signal data acquisition system are all connected to a computer.

The utility model has the following beneficial effects: under the control of the computer program, the utility model can move the three-dimensional electric precision translation table or the laser scanning device so that the laser can be focused on the position point in the xy plane, and the computer program controls the pulse laser light source to emit Laser pulse, the photoelectric detector converts the measured laser pulse signal into an electrical pulse to provide trigger signals for two sets of data acquisition systems, and the two sets of data acquisition systems collect and record the spectral signals of the spectrometer and the ultrasonic waves received by the ultrasonic transducer Signal. After completing point-by-point scanning of a preset area, the computer program processes the collected data to obtain photoacoustic microscopic images, fluorescent microscopic images, and fluorescent microspectral images of the sample. The utility model can obtain the forward or reverse photoacoustic signal through the signal switching switch. The scanning mode of the laser scanning device or the scanning mode of the three-dimensional precision electric translation stage (sample scanning mode) is selected through the computer program to obtain the scanning mode that meets the actual needs.

Description of drawings

Fig. 1 is a schematic diagram of the imaging analysis device described in the present invention.

Fig. 2 is a signal flow chart of the analysis device described in the present invention.

Fig. 3 is a schematic diagram of the water box.

detailed description

Below in conjunction with specific accompanying drawing, the utility model is further described.

As shown in Figures 1 to 3: the multifunctional photoacoustic, fluorescent microscopy and fluorescence spectrum imaging analysis device includes a computer 1, a three-dimensional electric precision translation stage 2, a laser scanning device 3, a pulsed laser light source 4, a beam splitter mirror 5, photodetector 6, ultrasonic signal data acquisition system 7, signal amplifier 8, thermal mirror 9, spectral signal data acquisition system 10, spectrometer 11, objective lens 12, water box 13, first ultrasonic transducer 14, second Ultrasonic transducer 15, signal switching switch 16, ultrasonic signal window 17, film 18, etc.

As shown in Figure 1, the multifunctional photoacoustic, fluorescent microscope and fluorescent spectrum imaging analysis device described in the utility model includes a three-dimensional electric precision translation stage 2 that can move in three directions of x, y and z. A water box 13 for placing samples is set on the precision translation stage 2; as shown in Figure 3, an ultrasonic signal window 17 is provided at the bottom of the water box 13, and the size of the ultrasonic signal window 17 is generally greater than the size of the sample. Ultrasonic signal window 17 covers the film 18 that can pass through ultrasonic waves; the first ultrasonic transducer 14 and the second ultrasonic transducer 15 that can adjust the orientation are respectively arranged at the bottom and side of the ultrasonic signal window 17. The ultrasonic transducer 14 and the second ultrasonic transducer 15 are respectively connected to the signal amplifier 8 through the signal switching switch 16, the signal amplifier 8 is connected to the ultrasonic signal data acquisition system 7, and the signal output terminal of the ultrasonic signal data acquisition system 7 is connected to the computer 1.

A laser scanning device 3 capable of scanning laser light in x and y directions is arranged above the three-dimensional electric precision translation stage 2, and an objective lens 12 is arranged between the laser scanning device 3 and the three-dimensional electric precision translation stage 2; One side of the laser scanning device 3 is provided with a hot mirror 9, a beam splitter mirror 5 and a pulsed laser light source 4 in sequence. The pulsed laser light emitted by the pulsed laser light source 4 is divided into two beams by the beam splitter mirror 5, and one beam is received by the photodetector 6. , a beam passes through the hot mirror 9 , the laser scanning device 3 and the objective lens 12 to reach the sample in the water box 13 on the three-dimensional electric precision translation stage 2 . The electrical signal output terminals of the photodetector 6 are respectively connected to the ultrasonic signal data acquisition system 7 and the spectral signal data acquisition system 10 . The spectral signal data acquisition system 10 is connected to the CCD camera of the spectrometer 11, and the spectrometer 11 collects the spectral information of back-facing fluorescence of each scanning point of the sample.

The above-mentioned three-dimensional electric precision translation stage 2, laser scanning device 3, pulsed laser light source 4, ultrasonic signal data acquisition system 7 and spectrum signal data acquisition system 10 are all connected to the computer 1, and the computer 1 controls their respective working states.

Aiming at the deficiencies of the traditional microscopic system and ultrasonic imaging technology, in order to obtain more characteristic information and higher spatial resolution of the sample, the utility model simultaneously records the ultrasonic signal generated by the laser pulse energy deposited in the sample and the laser in the sample. The fluorescence spectrum signal excited in the medium can simultaneously realize photoacoustic microscopic imaging, fluorescence microscopic imaging, and fluorescence microspectral imaging of the sample. In the utility model, under the control of a computer program, the movement of the three-dimensional electric precision translation stage or the laser scanning device can make the laser focus on the position point in the xy plane, and the computer program controls the pulsed laser light source to make it emit laser pulses, and the photoelectric detector The measured laser pulse signal is converted into an electrical pulse to provide trigger signals for two sets of data acquisition systems. The two sets of data acquisition systems collect and record the spectral signal of the spectrometer and the ultrasonic signal received by the ultrasonic transducer respectively. After completing point-by-point scanning of a preset area, the computer program processes the collected data to obtain photoacoustic microscopic images, fluorescent microscopic images, and fluorescent microspectral images of the sample.

The multi-functional photoacoustic, fluorescent microscopy and fluorescent spectrum imaging analysis method described in the utility model can realize three functions and has two scanning modes. The following steps are specifically adopted, as shown in Figure 2:

(1) Select the appropriate scanning method according to actual needs: in the control program of the computer 1, select the laser scanning device 3 to perform point-by-point light scanning or select the three-dimensional electric precision translation stage 2 to move to perform point-by-point sample scanning;

Selecting a suitable direction for ultrasonic signal collection: adjusting the spatial positions of the first ultrasonic transducer 14 and the second ultrasonic transducer 15 to a suitable direction;

After selecting the direction of ultrasonic signal acquisition, switch the signal switching switch 16 to the corresponding direction of the ultrasonic transducer (the direction of the first ultrasonic transducer 14 or the direction of the second ultrasonic transducer 15); adjust the position of the sample to focus the laser on In the area to be tested of the sample, the position of the corresponding ultrasonic transducer is precisely adjusted again so that the laser focus is in the center of the sound field detected by the ultrasonic transducer;

(2) The computer 1 program controls the laser scanning device 3 or the three-dimensional electric precision translation stage 2 to move to a predetermined position; the computer 1 sends a trigger signal to trigger the pulse laser light source 4 to send out laser pulses, and the photodetector 6 receives the laser pulse signals Converted into electrical pulse signals to trigger the ultrasonic signal data acquisition system 7 and the spectral signal data acquisition system 10; the laser pulse energy emitted by the pulsed laser light source 4 is deposited in the sample, and the sample absorbs the laser pulse energy and thermally expands to generate an ultrasonic signal. The transducer is converted into an electrical signal and amplified by the signal amplifier, and the ultrasonic signal data acquisition system 7 collects and records the ultrasonic signal sent by each scanning point of the sample; at the same time, the CCD camera in the spectrometer 11 synchronously collects the sample. For fluorescent spectral signals, the spectral signal data acquisition system 10 collects and records the fluorescent spectral information of the spectrometer 11 at each scanning point of the sample;

(3) Under the control of the computer 1 program, the laser scanning device 3 or the three-dimensional electric precision translation stage 2 scans point by point, repeats the data acquisition process of step (2) at each scanning point, and completes the measurement of all points in the area to be measured. data collection work;

(4) Ultrasonic signal data acquisition system 7 and spectral signal ultrasonic acquisition system 10 transmit all the collected data to computer 1, and computer 1 program processes all the data and gives the analysis images of photoacoustic microscopy, fluorescence microscopy and fluorescence spectrum of the sample .

The utility model has the following advantages: (1) The utility model can simultaneously collect the ultrasonic signal generated after the laser pulse energy is deposited in the sample, and the back-facing fluorescence spectrum signal generated by the sample under the excitation of the laser pulse, which can be obtained by computer processing Photoacoustic microscopic image, fluorescent microscopic image and fluorescent microspectral image of the sample; (2) The utility model can realize optical scanning or sample Scanning; (3) The utility model selects different signal sources through the signal switching switch, and can obtain forward or backward photoacoustic signals according to actual needs.

Claims (6)

1. a multi-functional optoacoustic, fluorescence microscopy and fluorescence spectrum imaging analysis device, it is characterized in that: including can at x, three-D electric precision translation stage (2) that y and z moves in three directions, laser scanning device (3), pulsed laser light source (4), spectrogrph (11), ultrasonic signal acquisition system (7), spectroscopic acquisition system (10), first ultrasonic transducer (14) and the second ultrasonic transducer (15), three-D electric precision translation stage (2) is arranged to place the water box (13) of sample, first ultrasonic transducer (14) and the second ultrasonic transducer (15) connect signal amplifier (8) respectively through signal shift switch (16), signal amplifier (8) connects ultrasonic signal data collecting system (7), at the laser scanning device (3) being provided above making laser move in x, y direction of described three-D electric precision translation stage (2), object lens (12) are set between laser scanning device (3) and three-D electric precision translation stage (2), heat mirror (9), beam splitter (5) and pulsed laser light source (4) is set gradually in the side of described laser scanning device (3), described spectral signature data acquisition system (10) connects the CCD camera of spectrogrph (11).

2. multi-functional optoacoustic as claimed in claim 1, fluorescence microscopy and fluorescence spectrum imaging analysis device, is characterized in that: the size of described ultrasonic signal window (17) is more than the size of sample.

3. multi-functional optoacoustic as claimed in claim 1, fluorescence microscopy and fluorescence spectrum imaging analysis device, it is characterized in that: be provided with ultrasonic signal window (17) in described water box (13) bottom, cover at ultrasonic signal window (17) and hyperacoustic thin film (18) can be passed through.

4. multi-functional optoacoustic as claimed in claim 1, fluorescence microscopy and fluorescence spectrum imaging analysis device, is characterized in that: described ultrasonic signal data collecting system (7) and spectral signature data acquisition system (10) connect the electrical signal of photodetector (6) respectively.

5. multi-functional optoacoustic as claimed in claim 1, fluorescence microscopy and fluorescence spectrum imaging analysis device, is characterized in that: described three-D electric precision translation stage (2), laser scanning device (3), pulsed laser light source (4), ultrasonic signal data collecting system (7) and spectral signature data acquisition system (10) are all connected with computer (1).

6. multi-functional optoacoustic as claimed in claim 1, fluorescence microscopy and fluorescence spectrum imaging analysis device, is characterized in that: described first ultrasonic transducer (14) and the second ultrasonic transducer (15) are separately positioned on ultrasonic signal window (17) bottom and sidepiece.