Automatic focusing system and method based on multi-depth plane microscope
An autofocus, depth plane technology, applied in microscopes, optics, instruments, etc., can solve the problems of increasing algorithm time and time-consuming, and achieve the effect of speeding up autofocus time
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Embodiment 1
[0060] hardware system such as figure 1 shown. The light source 1 is introduced into the Kohler illumination system 3 through the optical fiber 2, and the illuminated sample 6 is imaged on the main camera 11 through the objective lens 7 and the tube lens 9; Camera 13 on. For example, in this embodiment, the objective lens is Plan NA0.25 10x objective lens and Plan NA0.75 20x objective lens, and the beam splitting systems 10 and 12 are prisms (groups), which can be clamped and installed by machined parts. The splitting ratio of the spectroscopic systems 10 and 12 should be designed so that the brightness of the image formed by each depth plane is uniform, neither overexposed nor too dark; the spectroscopic system 12 needs to be designed so that the image size of each depth plane is basically the same. consistent, and have similar aberrations. In this embodiment, the beam splitting system 12 is a beam splitting prism. Taking this prism group as an example, it can be designed ...
Embodiment 2
[0065] hardware system such as figure 2 shown. The light source 1 is introduced into the Kohler illumination system 3 through the optical fiber 2, and the illuminated sample 6 is imaged on the main camera 11 through the objective lens 7 and the tube lens 9; Camera 13 on. The spectroscopic system 12 is a secondary grating, such as Figure 5 As shown, the secondary grating can be part of a Fresnel grating. The light passing through the secondary grating can exit from different angles through diffraction, such as Image 6 As shown, the 0-order and ±1-order outgoing light are emitted through the secondary grating, and different orders of light will image images with different focal depths on the camera.
[0066] The auxiliary camera 13 can simultaneously image three different focal planes. The motion control system 5 controls the motion platform 4 to move and scan in the x and y directions, and controls the objective lens 7 or the tube lens 9 to perform z-axis focus scanning...
Embodiment 3
[0069] hardware system such as image 3 shown. The light source 1 is introduced into the Kohler illumination system 3 through the optical fiber 2, and the illuminated sample 6 is imaged on the main camera 11 through the objective lens 7 and the tube lens 9; Camera 13 on. The light splitting system 12 is two mutually orthogonal secondary gratings, such as Figure 7 shown. The auxiliary camera 13 can simultaneously image 9 different focusing planes. The motion control system 5 controls the motion platform 4 to move and scan in the x and y directions, and controls the objective lens 7 or the tube lens 9 to perform z-axis focus scanning.
[0070] The autofocus scanning process and algorithm are the same as those in Embodiment 1.
[0071] In order to ensure the consistency and continuity of the focus during microscope scanning, curve fitting or artificial intelligence learning can be performed on the focus z value of the scanned position to predict the focus z value of the nex...
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