CN117801946A - Light stimulation system capable of being combined with multiple electrophysiological devices and application method thereof - Google Patents

Abstract

The invention discloses a light stimulation system that can be combined with a variety of electrophysiological equipment and a method of using it. The light stimulation system includes a computer, a projector, a base plate, a plurality of liftable pillars, a convex lens assembly and a light splitting prism. Elevating pillars are installed on the base plate, and the computer is connected to the projector and electrophysiological equipment. The projector is placed on one of the elevating pillars, and a convex lens assembly is installed on the upper end of the remaining elevating pillars. A plurality of sliding rods are installed transversely on the convex lens assembly, and a dichroic prism is installed on the sliding rod. The light stimulation system of the present invention is suitable for use with a variety of equipment and is not limited by instruments, making the system widely applicable. It can carry out related tests on living animals, isolated retinas, and optic cup organoids, and carry out research on visual systems. , to expand the functions of existing instruments.

Description

A light stimulation system that can be combined with multiple electrophysiological devices and its use method

Technical field

The present invention relates to the technical field of medical devices, and specifically to a light stimulation system that can be combined with multiple electrophysiological devices and a method of using the same.

Background technique

The formation of vision is that the photoreceptor cells on the retina in the eyeball convert external light signals into electrical signals. The electrical signals are transmitted to ganglion cells via bipolar cells, and the ganglion cells convert them into nerve signals and transmit them backward until they reach the cerebral cortex. To form vision. The complex visual nervous system is the necessary foundation to support the realization of visual functions. In recent years, in order to explore the encoding, decoding and information transmission mechanisms of neural networks in the visual system, the retinas of vertebrates and primates have been stimulated, combined with electrophysiological recordings and many other methods. technology to explore the deep mechanisms of visual neural networks. In existing research, the retina is used as the research object, and the neural response of the retina is induced by using light stimulation of a fixed wavelength and a single shape, and its response pattern is analyzed. However, the stimulation paradigm is simple and the stimulation parameters are fixed, which makes it impossible to fully verify the retina's ability to encode and decode arbitrary illumination stimuli in parallel.

Moreover, in the existing technology, a set of electrophysiological equipment is only paired with a set of light stimulation systems. For example, the polygon series modulators and spatial illuminators of MIGHTEX can only be used in conjunction with the patch clamp system; the light stimulation module STG4002-1.6A-opto of multi-channel systems can only be used in conjunction with the multi-electrode array MEA2100, and the light stimulation can only cover the entire chip area, and cannot select a specific location for stimulation; the light stimulation module of Axion Biosystems is integrated into the multi-electrode array system. It can be seen that a set of light stimulation systems are needed to be used in conjunction with the multi-electrode array and the patch clamp.

Summary of the invention

In order to solve the deficiencies of the above technical solutions, the object of the present invention is to provide a light stimulation system that can be used in conjunction with a variety of electrophysiological devices.

Another object of the present invention lies in the method of using the above-mentioned light stimulation system.

The object of the present invention is achieved through the following technical solutions.

A light stimulation system that can be used in conjunction with a variety of electrophysiological devices includes a computer, a projector, a base plate, a plurality of liftable supports, a convex lens assembly and a dichroic prism, wherein the liftable supports are mounted on the base plate, the computer is connected to the projector and the electrophysiological device, the projector is placed on one of the liftable supports, a convex lens assembly is mounted on the upper end of the remaining liftable supports, a plurality of slide bars are horizontally mounted on the convex lens assembly, and a dichroic prism is mounted on the slide bars.

In the above technical solution, four mounting holes are arranged on the convex lens assembly, and a sliding rod is installed in the mounting holes on the same horizontal line.

In the above technical solution, the bottom plate is a porous plate.

In the above technical solution, the number of the liftable pillars is 3, a support platform is installed on one of the liftable pillars, the projector is placed on the support platform, and the remaining two liftable pillars are placed on the support platform. A bracket is installed, and a convex lens assembly is installed on each bracket.

In the above technical solution, the convex lens assembly includes a convex lens and a convex lens bracket, the convex lens is embedded in the convex lens bracket, the convex lens bracket is rectangular, and a mounting hole is opened at each of the four right angles of the convex lens bracket, 2 Each of the convex lens brackets is equipped with a sliding rod in the installation hole on the same horizontal line.

In the above technical solution, the number of the sliding rods is 4, the 4 sliding rods are transversely parallel, 4 through holes are opened on the dichroic prism, and the 4 sliding rods are inserted into the 4 through holes. .

In the above technical solution, the liftable pillar includes a base and a sliding rod. The sliding rod is slidably installed in the base, and an adjustment knob is installed on the base.

The method of using the above-mentioned light stimulation system includes the following steps:

Step 1. Connect the electrophysiological equipment to the computer, use the software to record data, and set the recording and stimulation to start simultaneously;

Step 2, control the light stimulation parameters through the programming of the computer of the light stimulation system, generate a light stimulation image of any shape, speed, light intensity and color and transmit it to the projector;

Step 3. Use the adjustment button on the liftable pillar to adjust the height of the sliding rod so that the convex lens and the projector are on the same level. Move the convex lens assembly on the sliding rod to adjust the distance between the convex lens and the projector so that the light emitted by the projector After passing through the convex lens, the light path direction is changed through the dichroic prism, so that the light stimulation pattern on the projector is illuminated on the electrophysiological equipment containing the retina;

Step 4: The retina converts light signals into electrical signals, triggering the flow of ions inside and outside the optic ganglion cells, generating a potential difference on both sides of the membrane, putting the optic ganglion cells in an excited state, and the electrophysiological equipment detects the electrical signals.

In the above technical solution, the electrophysiological device is a patch clamp system or a multi-electrode array.

The advantages and beneficial effects of the present invention are:

The light stimulation system of the present invention is suitable for use with a variety of devices and is not limited by instruments, making the system widely applicable and capable of conducting relevant tests on living animals, isolated retinas, and optic cup organoids, conducting research on the visual system, and expanding the functions of existing instruments.

When the light stimulation system of the present invention is used in conjunction with an electrophysiological device, the light stimulation system can achieve synchronization with the data recording of the electrophysiological device through the HDMI interface of the projector and the USB interface of the electrophysiological device, achieving millisecond accuracy.

The light stimulation system of the present invention, when used in conjunction with electrophysiological equipment, can implement a variety of stimulation schemes through programming, including stimulation at different frequencies, different brightness, different colors, different shapes, different speeds and different times, and the stimulation parameters are flexible and adaptable. Adjustment can support the research of visual systems and expand the functions of existing instruments.

Description of drawings

Figure 1 is a schematic structural diagram of the light stimulation system of the present invention;

Figure 2 is a schematic structural diagram of the combination of the light stimulation system and the patch clamp system of the present invention;

FIG3 is a schematic diagram of the structure of the light stimulation system and the multi-electrode array system of the present invention;

Figure 4 is a schematic diagram of the stimulation scheme of different illumination parameters of the light stimulation system of the present invention;

Figures 5-8 are schematic diagrams of stimulation schemes of different shapes, sizes and colors of the light stimulation system of the present invention;

FIG9 is a schematic diagram of stimulation schemes of different motion directions and speeds of the light stimulation system of the present invention;

Figure 10 is a data diagram of the combination of the light stimulation system and the patch clamp system of the present invention;

FIG11 is a data diagram of the combination of the light stimulation system and the patch clamp system of the present invention;

Figure 12 is a data diagram of the light stimulation system of the present invention combined with a multi-electrode array;

Figure 13 is a data diagram of the light stimulation system of the present invention combined with a diaphragm multi-electrode array.

in,

1: computer, 2: projector, 3: base plate, 4: liftable support, 4.1: base, 4.2: sliding rod, 4.3: adjustment knob, 5: convex lens, 6: beam splitter prism, 7: electrophysiological equipment, 8: sliding rod, 9: bracket, 10: convex lens bracket, 11: support table.

Detailed ways

The technical solution of the present invention will be further described below with reference to specific embodiments.

Example 1

As shown in Figures 1 to 3, a light stimulation system that can be used in conjunction with a variety of electrophysiological devices includes a computer 1, a projector 2, a base plate 3, a plurality of liftable pillars 4, a convex lens assembly and a dichroic prism 6, wherein the liftable pillars 4 are fixedly mounted on the base plate 3 by screws, the computer 1 is connected to the projector 2 and the electrophysiological device 7, the projector 2 is placed on one of the liftable pillars 4, a bracket 9 is mounted on the upper end of the remaining liftable pillars 4, a convex lens assembly 5 is mounted on the bracket 9 (fixed with screws), four mounting holes are provided on the convex lens assembly, a slide bar 8 is installed in the mounting holes on the same horizontal line, and a dichroic prism 6 is installed on the slide bar 8.

Specifically, the bottom plate 3 is a porous plate, the number of the liftable pillars 4 is 3, a support platform 11 is installed on one of the liftable pillars 4, and the projector 2 is placed on the support platform 11. A bracket 9 is installed on the remaining two liftable pillars 4 (that is, there are two brackets 9), and a convex lens assembly is installed on each bracket 9 (that is, there are two convex lenses 5, installed through screws), The convex lens assembly includes a convex lens 5 and a convex lens bracket 10. The convex lens 5 is embedded in the convex lens bracket 10. The convex lens bracket 10 is rectangular, and a mounting hole is opened at each of the four right angles of the convex lens bracket 10. 2 Each of the convex lens brackets 10 is installed with a sliding rod 8 in the installation hole on the same horizontal line. The two sliding rods 8 below are fixed on the bracket 9 through buckles. The number of the sliding rods 8 is 4. 4 The sliding rods 8 are transversely parallel, four through holes are opened on the dichroic prism 6, and the four sliding rods 8 are inserted into the four through holes.

Specifically, the liftable pillar 4 includes a base 4.1 and a sliding rod 4.2. The sliding rod 4.2 is slidably installed in the base 4.1. An adjusting knob 4.3 is installed on the base 4.1, and the adjusting knob 4.3 is rotated. To realize the lifting and lowering of the sliding rod 4.2 in the base 4.1. As shown in Figure 2, the above-mentioned light stimulation system provided in this embodiment is used in conjunction with a patch clamp system:

The patch clamp system includes a dual-electrode patch clamp amplifier 700B, a digital-to-analog/analog-to-digital converter 1550A, a left- and right-hand electric micromanipulator MPC-385-2, and an inverted microscope IX73; the specification model of the projector 2 is OBEM3 A 250lm programmable LED projector; the specification model of the convex lens 5 is a plano-convex lens 5 with a radius of 50.8mm and a focal length of 100mm of GCL-010118A; psychtoolbox is pre-installed on the computer 1, and written through the EDITOR interface of the Matlab software Stimulation code, set parameters such as stimulation frequency, duration, color, shape, speed and recording synchronization, use HDMI data transmission cable to connect computer 1 and programmable LED projector to present stimulation, and at the same time pass data through the USB port of the patch clamp Connect the cable to the USB port of computer 1 to synchronize stimulation and data recording.

The method of using the above-mentioned light stimulation system in combination with the patch clamp system to test the accuracy of light stimulation of the retina includes the following steps:

First, Ames culture medium is prepared, and the retina removed from C57 mice is placed in the culture medium and oxygenated to maintain its activity. The retina is stimulated by light. The light stimulation parameters are controlled by computer programming to generate light stimulation images of any shape, speed, light intensity and color (as shown in Figures 4-9), and transmitted to the projector through the data transmission line. The adjustment button on the lifting support is adjusted to adjust the height of the sliding rod, and finally the convex lens and the projector are located at the same level. The convex lens assembly on the sliding rod is slid to adjust the distance between the convex lens and the projector, so that the light emitted by the projector passes through the convex lens. The distance between the two is different, and the size of the final light stimulation pattern is also different. After that, the light stimulation pattern changes the direction of the light path through the dichroic prism, so that the stimulation pattern is irradiated to the electrophysiological device containing the retina. The retina has a photoelectric conversion function, which converts light signals into electrical signals, which will trigger the flow of ions in and out of the retinal ganglion cells, and a potential difference is generated on both sides of the membrane, which makes the retinal ganglion cells in an excited state, and the electrophysiological device can detect the electrical signals. Use a data cable to connect the electrophysiological device and the computer, use the Multi Channel System software to record data, and set it to start recording and stimulation synchronously. As shown in FIG10 , the recording electrode forms a giant resistance seal with the retinal ganglion cell membrane, and the retinal ganglion cells are activated by light stimulation, and the action potential generated immediately after the light stimulation can be synchronously recorded.

It can be seen from the above tests that the light stimulation system of the present invention can be used in conjunction with the patch clamp system, and can accurately stimulate the area and size. The light stimulation and membrane potential recording are synchronized. The system has good stability. After integration, the software is easy to use and It has good flexibility and can set stimulation parameters arbitrarily, and run light stimulation and patch clamp recording at the same time.

The method of using the above-mentioned light stimulation system in conjunction with the patch clamp system to test the accuracy of light stimulation of optic cup organoids includes the following steps:

Well-cultured optic cup organoids, which have photoreceptor cells and optic ganglion cells, use a light stimulation system to stimulate the optic cup organoids, activate the optic ganglion cells, and simultaneously record the action potentials of the optic ganglion cells through patch clamp, as shown in Figure 11 Shown: the recording electrode forms a giant resistance seal with the optic cup organoid, and using light stimulation on it can simultaneously record the action potential generated at the moment of light stimulation or after stimulation. As can be seen from Figure 11, the light stimulation system of the present invention can perform very well. When used in conjunction with a patch clamp system, the combined software and hardware operations are easy, convenient, simple and flexible, and can meet the optical stimulation settings of various parameters. Combined with the patch clamp system, the instrument functions are expanded and the optical stimulation is included. Research methods were introduced into electrophysiological studies.

Therefore, from the above two sets of tests, it can be concluded that the light stimulation system provided by the present invention can be well combined with the patch clamp system, and the integrated software and hardware can meet the various parameters required for light stimulation experiments. set up. And in this example, the computer is connected to the projector and patch clamp system respectively through data lines, and then through programming, stimulation and recording are synchronized to achieve millisecond-level accuracy.

As shown in Figure 3, the optical stimulation accuracy test of the retina using the above-mentioned optical stimulation system provided by this embodiment and the multi-electrode array MEA2100 includes the following steps:

First, configure the Ames culture medium, place the retina removed from the C57 mouse in the culture medium, and ventilate it with oxygen to maintain its activity. Use the light stimulation of the present invention to stimulate the retina. The retina has a photoelectric conversion function and converts light signals into electrical signals. , will trigger the flow of ions inside and outside the optic ganglion cells, causing the cells to generate action potentials, and the extracellular spike signals will be recorded simultaneously through a multi-electrode array system. As shown in Figure 12, the light stimulation system of the present invention can accurately stimulate the retina, use a designed experimental paradigm for stimulation, and can simultaneously detect the discharge signals of the optic ganglion cells in the retina.

This embodiment provides a method for using the above-mentioned light stimulation system in combination with the multi-electrode array MEA2100 to test the light stimulation accuracy of optic cup organoids, including the following steps:

Well-cultured optic cup organoids, which have photoreceptor cells and retinal ganglion cells, are stimulated by a light stimulation system to activate retinal ganglion cells, and the extracellular spike signals of retinal ganglion cells are recorded by the multi-electrode array MEA2100, thereby realizing the combination of the light stimulation system and the multi-electrode array system. As shown in Figure 13: The optic cup organoid can be precisely stimulated, and the discharge signals of retinal ganglion cells in the optic cup organoid can be synchronously detected by using a designed experimental paradigm for stimulation.

In summary, the optical stimulation system of the present invention is suitable for use with multiple sets of equipment (patch clamp system and multi-electrode array system), and is not subject to instrument restrictions, making the system widely applicable and capable of performing isolated retinal and visual culture experiments. Conduct corresponding research on light stimulation related tests on cup organoids, expand the functions of existing instruments, and support neurobiology research on the regulation mechanism of visual neural circuits.

Figure 4 is a schematic diagram of the stimulation scheme of the light stimulation system of the present invention under different illumination parameters, first in the dark, then illumination for 1 s, resting in the dark for 1 s, illuminating for 2 s, resting for 1 s, illuminating for 3 s, resting for 1 s, and finally changing the illumination frequency according to By adjusting the light intensity using the sine wave method, the duration, intensity and frequency parameters of light stimulation can be flexibly adjusted, and the effects of different light stimulation parameters on experimental subjects can be explored in detail.

Figures 5 to 8 are schematic diagrams of the stimulation schemes of the light stimulation system of the present invention in different shapes, sizes and colors. It can not only realize simple shapes such as triangles, circles, squares, etc., but also realize natural images, and explore the effects of different shapes and different colors on light stimulation. Effects of Experimental Subjects.

Figure 9 is a schematic diagram of the stimulation scheme of the light stimulation system of the present invention with different movement directions and speeds. The red object is a moving bar, which moves along the diameter in different directions. It is divided into eight directions. The speed of the moving bar can also be adjusted to explore the experiment. Subject preferences for direction and speed.

The present invention has been illustratively described above. It should be noted that without departing from the core of the present invention, any simple deformations, modifications or other equivalent substitutions that can be made by those skilled in the art without spending creative efforts fall within the scope of this invention. protection scope of the invention.

Claims (9)

1. The utility model provides a but light stimulation system of multiple electrophysiology equipment of combination, its characterized in that includes computer, projecting apparatus, bottom plate, a plurality of liftable pillar, convex lens subassembly and beam splitter prism, the liftable pillar is installed on the bottom plate, the computer is connected projecting apparatus and electrophysiology equipment, the projecting apparatus is placed on one of them the liftable pillar, at the installation convex lens subassembly of remaining liftable pillar upper end transversely wear to adorn a plurality of slide bars on the convex lens subassembly wear to adorn a beam splitter prism on the slide bar.

2. The optical stimulation system according to claim 1, wherein 4 mounting holes are provided in the convex lens assembly, and a slide bar is inserted into the mounting holes on the same horizontal line.

3. The light stimulation system of claim 1, wherein the base plate is a porous plate.

4. The optical stimulation system according to claim 1, wherein the number of the liftable struts is 3, a support table is mounted on one of the liftable struts, the projector is placed on the support table, a bracket is mounted on each of the remaining 2 liftable struts, and a convex lens assembly is mounted on each of the brackets.

5. The optical stimulation system according to claim 1, wherein the convex lens assembly comprises a convex lens and a convex lens support, the convex lens is embedded in the convex lens support, the convex lens support is rectangular, each of four right angles of the convex lens support is provided with a mounting hole, and 2 mounting holes of the convex lens support on the same horizontal line are penetrated with a sliding rod.

6. The optical stimulation system according to claim 1, wherein the number of the sliding rods is 4,4 sliding rods are transversely parallel, 4 through holes are formed in the beam splitting prism, and 4 sliding rods are installed in the 4 through holes in a penetrating mode.

7. The light stimulation system of claim 1, wherein the liftable support post comprises a base and a sliding bar slidably mounted within the base, the base having an adjustment knob mounted thereon.

8. Use of the optical stimulation system according to any of claims 1-7, characterized in that it comprises the following steps:

step 1, connecting electrophysiological equipment with a computer, and recording data by using software, wherein the recording and the stimulation are synchronously started;

step 2, controlling the light stimulation parameters through the programming of a computer of the light stimulation system, generating light stimulation images with any shape, speed, illumination intensity and color, and transmitting the light stimulation images to a projector;

step 3, adjusting the height of the sliding rod through an adjusting button on the lifting support column so that the convex lens and the projector are positioned on the same horizontal plane, moving a convex lens component on the sliding rod, adjusting the distance between the convex lens and the projector, enabling light emitted by the projector to pass through the convex lens, and changing the direction of a light path through the beam splitting prism, so that a light stimulation pattern on the projector is irradiated on the electrophysiological equipment containing retina;

and 4, converting the optical signals into electric signals by the retina, inducing the ions of the ganglion cells to flow inside and outside, generating potential difference on two sides of the membrane, enabling the ganglion cells to be in an excited state, and detecting the electric signals by the electrophysiological equipment.

9. The method of use according to claim 8, wherein the electrophysiology is provided as a patch clamp system or a multi-electrode array.