A fully automatic fast focusing method for large field of view acquisition of microscopic digital images
A digital image and focusing method technology, which is applied in microscopes, instruments, optics, etc., can solve the time-consuming problem of sample collection and focusing, and achieve the effect of speeding up and improving the quality of collection
Active Publication Date: 2020-10-16
EAST CHINA NORMAL UNIV +1
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[0004] The purpose of the present invention is to provide a fully automatic fast focusing method for large field of view acquisition of microscopic digital images, aiming to solve the problem that the sample acquisition focusing process is very time-consuming in the prior art
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[0033] In the present invention, there are two ways of three-axis movement of X, Y, and Z: one is that the stage can move in three axes of X, Y, and Z, and the objective lens and camera are fixed; the other is that the stage can only move horizontally , The objective lens and camera move vertically.
[0034] In this embodiment, the microscopic digital image large field of view acquisition system is composed of a light source, a stage that can move in X, Y, and Z three axes, an electric rotating stage that can switch the object lens, and a camera. The structure diagram of the system is as figure 2 As shown, it includes but is not limited to: 201 light source, 202 electric stage, 203 switchable objective lens electric rotating stage, 204 camera. The relationship between the sample plane and the reference plane is as follows image 3 As shown, the sample plane on the glass slide and the reference plane often have a certain inclination angle.
[0035] The flow chart of the fully auto...
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The invention discloses a fully-automatic fast focusing method used for large-field-of-view collection of a microscopic digital image. The method comprises the steps of: shooting an overall image of asample on a glass slide, using an image algorithm to identify a sample contour, and obtaining a sample region; selecting three noncollinear positions with samples from the sample region, respectivelycarrying out Z-axis focusing operation on the three positions on the basis of a sharpness evaluation function, and recording height of each focal surface relative to a Z-plane zero-point position; obtaining a sample space plane equation according to the focal-surface height of the three positions; and then carrying out large-field-of-view collection of the microscopic digital image, calculating and obtaining focal-surface adjustment height, which corresponds to focusing under the positions, according to a center position of the collection region and the sample plane equation, and thus realizing fast focusing of the region. Compared with methods of adjusting focal surfaces on the basis of sharpness evaluation functions for all collection regions, the fully-automatic fast focusing method greatly increases collection speed while image collection quality is guaranteed.
Description
Technical field [0001] The invention belongs to the field of digital image acquisition technology and focusing technology, and in particular relates to an automatic fast focusing acquisition method for microscopic digital image acquisition with a large field of view. Background technique [0002] Microscopic observation of biological samples has always been very time-consuming and inefficient. In recent years, the automatic microscopy platform has gradually replaced the original manual microscopy. The fast-developing high-speed network technology, image technology and computer technology have provided a solid foundation for the collection of microscopic digital images, and also put forward requirements for the speed and quality of its collection. [0003] At present, in the microscopy system, because the sample plane and the reference plane on the glass slide often have a certain inclination angle, it is necessary to refocus every time a position is changed to ensure the collected...
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IPC IPC(8): G02B21/24G02B21/36
CPCG02B21/241G02B21/361G02B21/365
Inventor 徐怀奇刘洪英庄泉洁徐相华
Owner EAST CHINA NORMAL UNIV