CN102590067A - Loading device capable of utilizing common microscope to conduct blood cell phase imaging - Google Patents

Abstract

The invention relates to a biological cell phase microscope imaging device, in particular to a loading device capable of performing phase imaging of blood cells by using an ordinary microscope. The loading device includes a laser, a first prism whose coating surface is a first beam splitting surface and a first reflection surface, a second prism whose coating surface is a second reflection surface and a second beam splitting surface, and a prism with a stage Install the frame body. The invention can be installed on the two-dimensional adjustable stage of a common microscope, applying the principle of MZ interference, combining and interfering the light beam passing through the blood cell and the light beam not passing through the blood cell to form a miniature phase image, which is passed through the loaded common microscope Microscopically magnified imaging, the microscopically magnified image can be connected to a computer for phase unpacking and inversion to form a three-dimensional image of cells, which solves the problem that ordinary microscopes cannot image blood cell transparent bodies.

Description

A loading device capable of phase imaging of blood cells using an ordinary microscope

technical field

The invention relates to a biological cell phase microscope imaging device, in particular to a loading device capable of performing phase imaging of blood cells by using an ordinary microscope.

Background technique

The detection and classification of blood cell morphology has important applications in life sciences, clinical medicine, bioengineering and other fields; the field of blood cell morphology detection technology is a science and technology that crosses multiple technical fields, and optical microscopy is a major method and technology ;Due to its optical transparency, blood cells generally need to be stained in microscopy, and staining is easy to cause damage to the original state of cells, so phase microscopy has become the latest and most useful method; phase microscopy imaging is mainly applied The principle of light interference produces a phase image, and then unpacks and reconstructs the three-dimensional image of the sample. At present, there are some patented application technologies related to microscopes and phase microscopes, such as patented technology CN97108464.5 (stereo false color optical microscope device and its method) Using the point light source to illuminate, the microscope mechanical barrel is transformed, and the color separation grating, Fourier transform lens and four-dimensional adjustable rectangular filter are installed to form a deformed white light image information processing system, thereby realizing the object on the microscope. The direct observation of the false-color stereoscopic images modulated by the black-and-white gray scale difference and the phase change rate of the microstructure according to the oblique illumination, but this inventive technology cannot microscopically image transparent blood cells; another example is the patented technology CN2009

10086592.2 (a quantitative digital microscopic phase contrast imaging method) is implemented on a common digital optical microscope platform and a common personal computer. This technology collects experimental image data on the focal plane by manual or automatic focusing. Control the movement of the stage, collect experimental image data on the upper and lower staggered focal planes, and calculate the change rate of the intensity in the direction of the optical axis based on the image on the focal plane collected above and the two images on the upper and lower staggered focal planes. Solve the light intensity propagation equation to achieve quantitative phase recovery and microscopic phase contrast imaging, but this invention is only invented on the optical path of the microscope, but an invention on the algorithm; there is also foreign patent technology CN101727059 (A ) (Digital holographic micro scopic imaging method based on surface plasma resonance and microscope) This method is mainly based on a holographic display of phase and intensity under the two-beam interference method. Micro method; another example is the foreign patented technology US2009290156 (A1) (Spatial light interference microscopy and fourier transform light scattering for cell and tissue characterization) This technology is mainly a The cell phase imaging method based on the Fourier scattering method; these two foreign patented technologies are very advanced and professional, but the cost of the instrument is high, the purpose is single, and the application threshold is high; the present invention is based on ordinary microscopes Microscopic imaging function, according to the principle of light interference, a patented technology of loading device with stable performance, simple structure, low cost, simple operation and convenient installation is proposed. Features of Phase Imaging.

Contents of the invention

The purpose of the patent of the present invention is to provide a loading device that can be loaded on an ordinary microscope for phase imaging of blood cells.

The technical solution of the present invention is:

A loading device capable of performing phase imaging of blood cells using an ordinary microscope, the loading device is fixed on an adjustable stage of the microscope, and is characterized in that: the loading device includes a laser, and the coating surfaces are respectively the first beam splitting surface and the first The first prism on the reflective surface, the second prism on the second reflective surface and the second spectroscopic surface on the coated surface, and the prism mounting frame with a phase microscope stage; the horizontal central axis of the first prism is the same as the laser axis The axis and level are installed on the lower part of the prism mounting frame, and the first splitting surface of the first prism is located in the middle of the first reflection surface of the first prism and the laser; the second prism is stacked vertically and coaxially with the first prism, and installed on On the upper part of the mounting frame, the second reflective surface of the second prism is coaxial with the first beam splitting surface of the first prism in the vertical direction, and the second beam splitting surface of the second prism is coaxial with the first reflective surface of the first prism in the vertical direction The light transmission hole is placed on the top of the prism mounting frame, coaxial with the second splitting surface of the second prism and the first reflection surface of the first prism; the phase microscope stage is integrated with the prism mounting frame and is located on the second In the middle of the second light splitting surface of the prism and the first reflecting surface of the first prism; there is a detachable cover plate on the left side of the frame body, which is fixed on the prism mounting frame body through the cover plate fixing thread hole.

The laser light source emits a beam of light in the horizontal direction, which is transmitted forward by the first beam splitting surface of the first prism into two beams of light that continue to transmit horizontally and vertically upward, and the beam that continues to transmit horizontally meets the first reflection of the first prism in the forward direction The surface is redirected to a vertically upward beam, which passes through the cell sample placed on the phase microscope stage and becomes the object light, enters the second beam splitting surface of the second prism, and is directly transmitted to the objective lens of the microscope system; by the first prism The first beam splitting surface is divided into the light beam that continues to transmit vertically upwards to the second reflective surface of the second prism along the vertical direction, and after reflection, the light beam in the horizontal direction is transmitted to the second beam splitting surface of the second prism, and becomes the reference light after reflection. The direction is vertical, and it enters the objective lens of the microscope system coaxially and in the same direction as the object light; the object light and the reference light meet and interfere through the microscopic magnification system composed of the microscope objective lens and the eyepiece to form a phase image of the cell sample; the whole device is installed in the microscope On the two-dimensional adjustable stage, clear cell phase images can be obtained after horizontal positioning and vertical focusing.

The prism is made of quartz glass, the reflective surface is the incoming beam surface of the coating total reflection surface; the splitting surface is the incoming beam surface of the coating semi-reflective and semi-transmissive, each of which is 50% of the incident light intensity; the reflecting surface and the splitting surface are strictly Parallel, each intersects with the bottom surface of the prism mounting frame at an angle of 45° to ensure the effect of M-Z interference optical path.

The ordinary microscope should be equipped with a two-dimensional adjustable stage. If it is not configured, the finished product can be purchased; when the device of the present invention is installed on the stage, it can be fine-tuned and positioned in two dimensions to ensure that the object light and reference light are consistent with the microscope light. Axis coaxial, to better realize the goal of the present invention.

When an ordinary microscope performs general microscopic magnification imaging, its sample only needs to be placed on the surface of the transmission hole of the device of the present invention, that is, the ordinary microscope stage can realize the general microscope function; when phase microscopy is required, the cell sample can be placed on the surface The phase microscopy function of the cell can be realized at the phase microscopy stage of the device.

Compared with related technologies, the patent of the present invention has the following remarkable advantages: 1. The patented technology that the device of the present invention can be loaded on an ordinary microscope to realize phase microscopy has not been reported, and is original; 2. The characteristics of the traditional optical path of M-Z guarantee the The high reliability and high stability of the device of the invention; 3. The double prism structure shows the simplicity of the overall structure and the convenience of operation and debugging; 4. The loading method of the device of the present invention not only ensures the function of the original microscope, but also increases the The function of the phase microscope realizes a machine with dual functions; 5, the device of the present invention effectively utilizes the parts and functions thereof of microscopic magnification, digital imaging, longitudinal focusing and plane two-dimensional positioning of the original microscope, therefore, the instrument cost Low, wide range of applications.

Description of drawings

Fig. 1 is a schematic structural view of a dual-purpose loading device for changing an ordinary microscope to a cell phase microscope according to the present invention;

Fig. 2 is a schematic diagram of the M-Z light interference principle of a dual-purpose cell phase microscope changed from an ordinary microscope to that of the present invention;

1. Laser 2. The first splitting surface of the first prism 3. The second reflecting surface of the second prism 4. The second splitting surface of the second prism 5. The first reflecting surface of the first prism 6. Light transmission hole 7. Prism positioning adjustment column 8. Prism mounting frame body 9. Cover plate fixing thread hole 10. Laser positioning thread hole 11. Frame body fixing thread hole 12. Phase microscope stage 13. Microscope adjustable stage 14. Ordinary display Micro stage 15. Microscope system of ordinary microscope 16. Cell samples.

Detailed ways

In combination with Fig. 1 and Fig. 2, a dual-purpose loading device for changing an ordinary microscope to a cell phase microscope in the present invention is an M-Z optical path interference system, including a laser 1, which is respectively coated into a first light splitting surface 2 and a first reflection surface 5 It consists of the first prism, the second prism coated to form the second reflective surface 3 and the second beam splitting surface 4 respectively, the phase microscope stage 12 , the transmission hole 6 and the frame 8 .

The collimated light beam emitted by the laser 1 is divided into two beams of light by the first light splitting surface 2 of the first prism, the horizontally forward and vertically upwardly transmitted light beams, and the horizontally transmitted light beam meets the first reflective surface of the first prism in the forward direction 5 is redirected to a vertically upward light beam, which becomes object light after passing through the cell sample 16 and enters the second light splitting surface 4 of the second prism, and is directly transmitted to the ordinary microscope system 15; The light beam transmitted vertically upwards goes to the second reflective surface 3 of the second prism along the vertical direction, and after reflection, the light beam in the horizontal direction is transmitted to the second light splitting surface 4 of the second prism, and becomes the reference light after reflection and is redirected to the vertical direction, Enter the ordinary microscope system 15 coaxially and in the same direction as the object light; the object light and the reference light meet and interfere through the microscope system 15 to form a phase image of the cell sample; the adjustment function of the prism positioning adjustment column 7 ensures that each reflection surface and light splitting surface are parallel to each other; A laser is installed in the laser positioning wire hole 10, which is used to ensure that the optical axis of the laser 1 is coaxial with the first beam splitting surface 2 of the first prism and the center of the first reflective surface 5 of the first prism; the cover plate fixing wire hole 9 is used for The fixing of the frame body cover plate; the whole loading device is installed on the two-dimensional adjustable stage 13 of the microscope through the frame body fixing thread hole 11, and the two-dimensional adjustable stage 13 is adjusted to adjust the coherent light beam and the light exit hole 6 of the device. The optical axis of the microscope is coaxial, and then a clear cell phase image can be obtained after the vertical focus of the common microscope system 15; the phase microscopic stage 12 and the common microscopic stage 14 are used for the phase and common phase of the device of the present invention The object of the microscopic pattern, so as to finally realize the object of the present invention.

Claims (2)

1. A loading device that can carry out blood cell phase imaging using a common microscope, the loading device is fixed on the adjustable stage of the microscope, it is characterized in that: the loading device includes a laser, and the coating surface is respectively the first beam splitting surface and the The first prism on the first reflective surface, the second prism whose coating surface is the second reflective surface and the second beam splitting surface respectively, the prism mounting frame body with the phase microscope stage; the horizontal central axis of the first prism and the laser The axis is coaxial and installed horizontally on the lower part of the prism mounting frame, and the first splitting surface of the first prism is located in the middle of the first reflective surface of the first prism and the laser; the second prism is stacked vertically and coaxially with the first prism, On the upper part of the prism mounting frame, the second reflective surface of the second prism is coaxial with the first light splitting surface of the first prism in the vertical direction, and the second light splitting surface of the second prism is vertical to the first reflective surface of the first prism. Coaxial; the light transmission hole is placed on the top of the prism mounting body, coaxial with the second splitting surface of the second prism and the first reflecting surface of the first prism; the phase microscope stage is integrated with the prism mounting body and located In the middle of the second splitting surface of the second prism and the first reflecting surface of the first prism; there is a detachable cover plate on the left side of the frame body, which is fixed on the prism mounting frame body through the cover plate fixing thread hole. 2. A loading device capable of performing phase imaging of blood cells using an ordinary microscope as claimed in claim 1, wherein the prism is made of quartz glass, and the reflection surface is the incoming beam surface of the coating total reflection surface; the beam splitting surface It is coated semi-reflective and semi-transmissive facing beam surfaces, each of which is 50% of the incident light intensity; the reflective surface and the splitting surface are strictly parallel, and each intersects with the bottom surface of the prism mounting frame at a 45° angle.

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