CN106610522A - Three-dimensional microscopic imaging device and method - Google Patents

Abstract

The invention discloses a three-dimensional microscopic imaging device and method. The device sequentially comprises an LED array, a sample carrying table, an objective lens, a red-blue color filter, a lens and a camera from the bottom to the top, and also comprises a computer and a red-blue stereo glasses. The center of the LED array is located on the central axis of the objective lens and the lens. The LED array and the camera are respectively connected with the computer. The method comprises the steps: placing a to-be-imaged sample on the sample carrying table; enabling the computer to transmit a control signal to the LED array, and enabling the LED array to generate white circular transparent light, wherein the light passes through the sample carrying table to be collected by the objective lens, and then is amplified for imaging, and finally enters the lens through the red-blue color filter; enabling the computer to drive the lens to carry out the sampling of a sample image passing through the lens, and enabling the camera to input the collected sample image into the computer through a data line for display; and carrying out the three-dimensional microscopic observation of the sample image displayed by the computer through the red-blue stereo glasses. The device is simple in structure, is low in cost, is good in effect, and can achieve the clear observation of the three-dimensional structure of a sample.

Description

A stereomicroscopic imaging device and method

technical field

The invention belongs to the technical field of optical microscopic imaging, in particular to a stereoscopic microscopic imaging device and method.

background introduction

As a general-purpose observation and measurement instrument, the microscope has long been used in all walks of life around us. It is the "materialized judge" of today's society and a "key to explore the microscopic world". As human beings observe the material world towards the micron and nanometer scales, the need for three-dimensional observation of tiny objects promotes the development of stereomicroscopic imaging technology. Stereo microscopes can also be called stereo microscopes or dissecting microscopes. They are widely used in various scientific research activities such as biology, chemistry, physics, metallurgy, brewing, and medicine, and have a significant impact on human development and progress.

The core idea of the existing traditional stereo microscope is to simulate the binocular parallax of human eyes to form stereo imaging. Human perception of the depth of nature is caused by the slight difference between the left and right eyes viewing the real world. There is a distance of 4-6cm between the human eyes, so in fact, when we see objects, the images in the two eyes are different. The left eye sees more components on the left side of the object, and the right eye sees more components on the right side of the object. After these two images are synthesized by the brain, the front, back, and distance of the object can be distinguished, thereby producing stereoscopic vision. What we see is an image with depth of field. People's three-dimensional stereoscopic vision of objects is produced by binocular parallax, so all optical devices or structures that can cause human eyes to produce binocular parallax can produce three-dimensional stereoscopic vision. Traditional stereo microscopes use a common primary objective lens to image the object, then separate them through two groups of intermediate objective lens groups with adjustable magnification, and then form images through eyepieces with an angle of 12 degrees to 15 degrees, so that the left and right eyes can be provided with different angles. After comprehensive processing by the brain, what the human eyes see is a stereoscopic image with depth of field.

The advantages of traditional stereo microscopes are obvious: easy to use and easy to operate. But the problem that it exists also can't make up for: (1) the included angle of the tube lens of two eyepieces is 12 degree～15 degree, and the body angle of view of such microscope mass production is certain. However, in real life, due to individual differences in the interpupillary distance of the human eye, the stereoscopic viewing angle of the human eye may not always be within this angle range, which will lead to unsatisfactory personal viewing effects; (2) The dual-channel optical path not only makes the instrument bulky but also costs a lot (3) The depth of focus cannot be adjusted, and the observer cannot independently control the observation thickness of the observed sample; (4) The imaging quality is poor, and the detailed information of the object cannot be viewed.

Contents of the invention

The object of the present invention is to provide a three-dimensional microscopic imaging device and method with simple structure, low cost and good effect, so as to realize clear viewing of the three-dimensional structure of a sample.

The technical solution that realizes the object of the present invention is: a kind of stereomicroscopic imaging device, comprises LED array, sample stage, objective lens, red and blue color filter, lens, camera, computer and red and blue stereoscopic glasses; Wherein LED array, The sample stage, objective lens, red and blue color filters, lens, and camera are arranged in sequence from bottom to top, and the center of the LED array is located on the central axis of the objective lens and lens; both the LED array and the camera are connected to the computer;

The sample to be imaged is placed on the sample stage, and the computer sends a control signal to the LED array to make the LED array generate a white circular illumination light with a radius of R pixels. The illumination light passes through the sample stage and is collected by the objective lens. The objective lens magnifies the collected illumination light and enters the lens through the red and blue color filters. The computer drives the camera to sample the sample image passing through the lens. Stereoscopic glasses perform stereomicroscopic observation of sample images displayed on a computer.

Preferably, the LED array uses a P4 series LED array, and the P4 series LED array provides seven colors of illumination light: red, green, blue, cyan, pink, yellow, and white light. The present invention uses white illumination light; the P4 series LED array The size of the cell board is 128mm*128mm, the number of pixels is 32*32, and the pixel pitch is 4mm. Each pixel can be lighted up individually.

Preferably, the distance between the LED array and the upper surface of the sample stage is 75-85 mm.

Further, the red and blue color filter is a circular light-transmitting film, and the filter is divided into two semicircles, and the two semicircles are respectively a red filter and a blue filter; the red and blue color filters The film is set on the back focal plane of the objective lens to separate the image formed by the objective lens.

Further, the CCD lens of the camera is located on the back focal plane of the lens.

Further, the red-blue anaglyph glasses have the same color as the red-blue color filter.

A stereomicroscopic imaging method, the steps are as follows:

Step 1. Place the sample to be imaged on the sample stage, and the computer sends a control signal to the LED array to make the LED array generate a white circular illumination light with a radius of R, where R is the number of pixels;

Step 2. The illumination light generated in step 1 passes through the sample stage and is collected by the objective lens. The objective lens magnifies the collected illumination light and then enters the lens through the red and blue color filters;

Step 3, the computer drives the camera to sample the sample image passing through the lens, and the camera inputs the collected sample image into the computer through the data line for display;

Step 4: Perform stereomicroscopic viewing of the sample image displayed by the computer through red and blue stereoscopic glasses, and at the same time adjust the radius R of the white circular illumination light generated by the LED array through the computer, and change the illumination numerical aperture angle to realize the different thickness layers of the sample. imaging.

Further, in step 4, the radius R of the white circular illumination light generated by the LED array is adjusted by the computer, and the illumination numerical aperture angle is changed to realize imaging of layers of different thicknesses of the sample, wherein the illumination numerical aperture angle θ _NA and the circular illumination light The relationship between the radius R is as follows:

θ _NA =arctan R/H

Wherein, H is the distance between the LED array and the upper surface of the sample stage.

Compared with the prior art, the present invention has significant advantages in that: (1) by adding red and blue color filters on the rear focal plane of the microscope objective lens, thereby replacing the dual optical path of the traditional stereo microscope, the system design is simplified, and the imaging system Simple and cost-effective; (2) By using LED arrays as the illumination source, the focal depth of the microscope can be flexibly adjusted; (3) By transferring the images collected by the camera to the computer display screen, it is possible to realize the three-dimensional observation of the sample. Clear view of the structure; (4) With the light source of the system itself, it is less affected by external interference, and the restriction of observation conditions is reduced, and it can be viewed under a dark background.

Description of drawings

Fig. 1 is a schematic structural diagram of a stereomicroscopic imaging device of the present invention.

Fig. 2 is a schematic diagram of the white circular illumination light generated by the LED array in the present invention.

Fig. 3 is a schematic structural view of the red and blue color filters in the present invention.

Fig. 4 is a sample diagram displayed by the computer in the embodiment of the present invention.

detailed description

The present invention will be described in further detail below in conjunction with the accompanying drawings and specific embodiments.

In conjunction with Fig. 1, the stereomicroscopic imaging device of the present invention comprises an LED array 1, a sample stage 2, an objective lens 3, a red and blue color filter 4, a lens 5, a camera 6, a computer 7 and red and blue stereoscopic glasses 8; wherein the LED Array 1, sample stage 2, objective lens 3, red and blue color filter 4, lens 5, and camera 6 are arranged in sequence from bottom to top, and the center of LED array 1 is located on the central axis of objective lens 3 and lens 5; LED array 1 and camera 6 are all connected with computer 7;

The sample to be imaged is placed on the sample stage 2, and the computer 7 sends a control signal to the LED array 1, so that the LED array 1 generates a white circular illumination light with a radius of R pixels, as shown in Figure 2, the illumination light Through the sample stage 2, it is collected by the objective lens 3, the objective lens 3 magnifies the collected illumination light and then enters the lens 5 through the red and blue color filter 4, and the computer 7 drives the camera 6 to sample the sample image passing through the lens 5 , the camera 6 inputs the sample image collected by the computer 7 through the data line for display, and the sample image displayed by the computer 7 is viewed through a stereoscopic microscope through the red and blue stereoscopic glasses 8 .

Preferably, the LED array 1 adopts a P4 series LED array, and the P4 series LED array provides seven colors of illumination light of red, green, blue, cyan, pink, yellow, and white light, and the present invention adopts a white illumination light source; the P4 series LED The size of the unit board of the array is 128mm*128mm, the number of pixels is 32*32, and the pixel pitch is 4mm. Each pixel can be individually lit.

Preferably, the distance between the LED array 1 and the upper surface of the sample stage 2 is 75-85 mm.

Further, as shown in Figure 3, the red and blue color filter 4 is a circular light-transmitting film, and the filter is divided into two semicircles, and the two semicircles are respectively the red filter and the blue filter. Light sheet; the red and blue color filter 4 is arranged on the rear focal plane of the objective lens 3, and separates the image formed by the objective lens 3.

Further, the CCD lens of the camera 6 is located on the back focal plane of the lens 5 .

Further, the red and blue anaglyph glasses 8 and the red and blue color filter 4 have the same color.

Stereomicroscopic imaging method of the present invention, the steps are as follows:

Step 1, place the sample to be imaged on the sample stage 2, and the computer 7 sends a control signal to the LED array 1, so that the LED array 1 generates a white circular illumination light with a radius of R, where R is the number of pixels;

Step 2, the illumination light generated in step 1 passes through the sample stage 2 and is collected by the objective lens 3, and the objective lens 3 magnifies and forms the collected illumination light and then enters the lens 5 through the red and blue color filters 4;

Step 3, the computer 7 drives the camera 6 to sample the sample image passing through the lens 5, and the camera 6 inputs the collected sample image through the data line to the computer 7 for display;

Step 4: Perform stereomicroscopic viewing of the sample image displayed by the computer 7 through the red and blue stereoscopic glasses 8, and at the same time adjust the radius R of the circular illumination light generated by the LED array 1 through the computer 7, and change the illumination numerical aperture angle to achieve different samples. Imaging of a thick layer, where the relationship between the illumination numerical aperture angle θ _NA and the radius R of the circular illumination light is as follows:

θ _NA =arctan R/H

Wherein, H is the distance between the LED array 1 and the upper surface of the sample stage 2 .

Example 1

The device of the invention and the steps for achieving stereo viewing of samples will be described in detail below in conjunction with the accompanying drawings.

(1) Introduce this inventive device in detail in conjunction with accompanying drawing:

In conjunction with Fig. 1, the stereomicroscopic imaging device of the present invention comprises a P4 series LED array 1, a sample stage 2, an objective lens 3, a red and blue color filter 4, a lens 5, a camera 6, a computer 7 and red and blue stereoscopic glasses 8 . The P4 series LED array 1 is installed under the sample stage 2, about 80 mm away from the upper surface of the sample stage 2, and its center is located on the central axis of the objective lens 3 and the lens 5. The computer 7 controls the P4 series LED array 1 to generate white illumination light with the required radius R (here, the radius R refers to the number of pixels), and the generated white illumination light passes through the sample stage 2 and is collected by the objective lens 3, and then undergoes amplification processing After imaging, the light is split by the red and blue color filters 4, and then further imaging processing by the lens 5, and the camera 6 is driven to collect pictures under the control of the computer 7. The computer 7 is connected with the P4 series LED array 1 and the camera 6, and its control is realized through a software system.

The P4 series LED array 1 can provide seven colors of illumination light: red, green, blue, cyan, pink, yellow and white. P4 series LED array 1 (unit board size 128mm*128mm, number of pixels 32*32, pixel pitch 4mm) is placed directly under the sample stage 2, the distance H from the upper surface of the stage is generally about 80mm, and The center of the P4 series LED array 1 is on the optical axis of the microscope optical system. Each pixel can be lighted up individually. The radius R and color of the circle displayed on the P4 series LED unit board are controllable, and the coordinates of the center of the circle are set at the center of the LED array, as shown in Figure 2. The radius R and the color of the lighting source are controlled by the computer 7, and only white light is used in this embodiment. The red and blue filter 4 is placed on the rear focal plane of the microscope objective lens, so that the image can be split, as shown in FIG. 3 . The lens 5 is a barrel lens, which realizes further magnification and processing of the image. The camera 6 collects images under the control of the computer 7, and displays them on the display screen of the computer 7, and the observer wearing the red and blue stereoscopic glasses 8 can realize stereo viewing of the sample.

(2) The specific steps for realizing the viewing of the three-dimensional object presented by the device include:

Step 1, place the sample to be imaged on the sample stage 2, and the computer 7 sends a control signal to the LED array 1, so that the LED array 1 generates a white circular illumination light with a radius of R, where R is the number of pixels;

Step 2, the illumination light generated in step 1 passes through the sample stage 2 and is collected by the objective lens 3, and the objective lens 3 magnifies and forms the collected illumination light and then enters the lens 5 through the red and blue color filters 4;

Step 3, the computer 7 drives the camera 6 to sample the sample image passing through the lens 5, and the camera 6 inputs the collected sample image through the data line to the computer 7 for display;

Step 4: Perform stereomicroscopic viewing of the sample image displayed by the computer 7 through the red and blue stereoscopic glasses 8, and at the same time adjust the radius R of the circular illumination light generated by the LED array 1 through the computer 7, and change the illumination numerical aperture angle to achieve different samples. Imaging of a thick layer, where the relationship between the illumination numerical aperture angle θ _NA and the radius R of the circular illumination light is as follows:

θ _NA =arctan R/H

Wherein, H is the distance between the LED array 1 and the upper surface of the sample stage 2, which is 80 mm in this embodiment.

Fig. 4 is a display image of a diffractive optical element sample captured by the device, which is displayed on a computer monitor, and a clear stereoscopic image can be observed by wearing red and blue stereoscopic glasses.

In summary, the present invention replaces the dual optical paths of a traditional stereomicroscope by adding red and blue color filters to the rear focal plane of the microscope objective lens, which simplifies system design, reduces costs, and improves imaging quality; The size is 128mm*128mm, the number of pixels is 32*32, and the pixel pitch is 4mm) is used as the illumination source to realize the flexible adjustment of the focal depth of the microscope. The system itself has its own light source, which is less affected by external interference; The display screen can realize the clear viewing of the three-dimensional structure of the sample.

Claims (8)

1. a kind of stereo micrography imaging device, it is characterised in that including LED array (1), sample stage (2), Object lens (3), red blue light filter (4), lens (5), camera (6), computer (7) and red blue anaglyph spectacleses (8)； Wherein LED array (1), sample stage (2), object lens (3), red blue light filter (4), lens (5), camera (6) set gradually from bottom to up, and LED array (1) is centrally located at object lens (3), the central shaft of lens (5) On line；LED array (1) and camera (6) are connected with computer (7)；

Sample to be imaged is placed in into sample stage (2), computer (7) sends control letter to LED array (1) Number, make LED array (1) produce the white circle illumination light that radius is R pixel, the illumination light is carried through sample Thing platform (2) is collected by object lens (3), and object lens (3) are amplified after imaging the illumination light of collection through red blue colour filter Piece (4) is incident to lens (5), and computer (7) drives camera (6) to carry out the sample image through lens (5) The sample image for gathering is shown by sampling, camera (6) through data wire input computer (7), by red blue vertical Body glasses (8) carry out stereomicroscopy viewing to the sample image that computer (7) shows.

2. stereo micrography imaging device according to claim 1, it is characterised in that the LED array (1) Using P4 series LED arrays, P4 series LEDs array provide redness, green, blueness, cyan, pink colour, yellow, The illumination light of seven kinds of colors of white light, the present invention adopt white illumination light；The unit board size of P4 series LED arrays is 128mm*128mm, number of pixels are 32*32, pel spacing 4mm, and each pixel individually can light.

3. stereo micrography imaging device according to claim 1, it is characterised in that the LED array (1) It is 75～85mm apart from the distance of sample stage (2) upper surface.

4. stereo micrography imaging device according to claim 1, it is characterised in that the red blue light filter (4) For circular light transmission piece, the optical filter is divided into two semicircles, and two semicircles are respectively Red lightscreening plate and blue filter Piece；Red blue light filter (4) is arranged on the back focal plane of object lens (3), will be object lens (3) imaging separate.

5. stereo micrography imaging device according to claim 1, it is characterised in that the CCD of the camera (6) Camera lens is located on the back focal plane of lens (5).

6. stereo micrography imaging device according to claim 1, it is characterised in that the red blue anaglyph spectacleses (8) With the solid colour of red blue light filter (4).

7. a kind of stereo micrography imaging method based on stereo micrography imaging device described in claim 1, it is characterised in that Step is as follows：

Sample to be imaged is placed in sample stage (2) by step 1, and computer (7) is sent to LED array (1) Control signal, makes LED array (1) produce the white circle illumination light that radius is R, numbers of the wherein R for pixel；

Step 2, the illumination light that step 1 is produced are collected by object lens (3) through sample stage (2), and object lens (3) will The illumination light of collection is incident to lens (5) through red blue light filter (4) after being amplified imaging；

Step 3, computer (7) drive camera (6) to sample the sample image through lens (5), camera (6) The sample image of collection is shown through data wire input computer (7)；

Step 4, carries out stereomicroscopy viewing to the sample image that computer (7) shows by red blue anaglyph spectacleses (8), Adjust the radius R of circular illumination light produced by LED array (1) simultaneously by computer (7), change illumination numerical Angular aperture is realizing the imaging to sample different-thickness layer.

8. stereo micrography imaging method according to claim 7, it is characterised in that by calculating described in step 4 Machine (7) adjusts the radius R of white circle illumination light produced by LED array (1), change illumination numerical aperture angle with Realize the imaging to sample different-thickness layer, wherein illumination numerical aperture angle θ_NAAnd the radius R of circular illumination light between Relation is as follows：

θ_NA=arctan R/H

Wherein, H is distance of the LED array (1) apart from sample stage (2) upper surface.