CN110794549B - Automatic focusing method, blood cell analyzer, and computer-readable storage medium - Google Patents
Automatic focusing method, blood cell analyzer, and computer-readable storage medium Download PDFInfo
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Abstract
The invention discloses an automatic focusing method, a blood cell analyzer and a computer readable storage medium, wherein the method comprises the following steps: resetting after starting; downwards moving the objective table by a first preset amplitude, and determining the next direction according to the image definition before and after movement; moving the objective table in a determined direction by a first preset amplitude, and determining the next direction according to the image definition before and after movement, the movement direction and a preset local extreme value threshold; moving the objective table in a determined direction by a second preset amplitude, and determining whether a third preset amplitude is adopted according to the definition of images before and after movement and a preset local extremum threshold; if not, continuing to move the objective table by a second preset amplitude; if so, adopting a third preset amplitude to move for a preset number of times in a direction opposite to the moving direction of the last second preset amplitude, and acquiring a focused image according to the image after each movement and the image after the last second preset amplitude movement. The method solves the problem that the real focal plane is not easy to find when the focal plane is searched by a climbing method.
Description
Technical Field
The present invention relates to the field of blood cell analyzers, and in particular, to an automatic focusing method, a blood cell analyzer, and a computer-readable storage medium.
Background
In the process of focusing a microscope, a climbing method is mainly adopted to search a focal plane at present. However, due to the unevenness of the surface of the sample plate and noise, the focal plane is searched by using the traditional climbing method, and the focal plane is easy to fall into a trap with a local extreme value, so that the real focal plane is not easy to find.
Disclosure of Invention
The invention mainly aims to provide an automatic focusing method, a blood cell analyzer and a computer readable storage medium, and aims to solve the problem that a real focal plane is not easy to find by adopting a climbing method to search the focal plane in the prior art.
To achieve the above object, the present invention provides an auto-focusing method, comprising the steps of:
starting reset according to a starting instruction input by a user, and executing a first moving operation of moving the objective table downwards according to a first preset amplitude;
acquiring the definition of images before and after the first moving operation to determine the moving direction of a second moving operation executed in a first preset amplitude, and executing the second moving operation according to the determined moving direction to acquire the definition of the images before and after the second moving operation;
determining the moving direction of a third moving operation executed in a second preset amplitude according to the definition of the images before and after the second moving operation, the moving direction of the second moving operation and a preset local extremum threshold, executing the third moving operation according to the moving direction, and determining whether to change the amplitude of the moving operation to a third preset amplitude smaller than the second preset amplitude according to the definition of the images before and after the third moving operation and the preset local extremum threshold;
if not, determining the moving direction of the next third moving operation according to the definition of the image before and after the third moving operation, a preset local extremum threshold and the accumulated continuous moving times which comprise the third moving operation and have the same moving direction as the third moving operation, and returning to the step of executing the third moving operation according to the moving direction;
if so, continuously executing the fourth moving operation for a first preset number of times in a third preset amplitude and the opposite moving direction of the last third moving operation, and comparing the image after the fourth moving operation executed according to the third preset amplitude each time with the image after the last third moving operation to obtain the focused image.
Optionally, the step of obtaining the sharpness of the image before and after the first moving operation to determine the moving direction of the second moving operation performed at the first preset magnitude includes:
judging whether the definition of the image after the first moving operation is greater than that of the image before the first moving operation;
if so, determining that the moving direction of the second moving operation is downward;
if not, determining that the moving direction of the second moving operation is upward.
Optionally, the step of determining, according to the sharpness of the image before and after the second moving operation, the moving direction of the second moving operation, and the preset local extremum threshold, the moving direction of the third moving operation performed at the second preset amplitude includes:
obtaining a local extreme value of the image after the second moving operation through a preset formula according to the definition of the image before and after the second moving operation;
judging whether the local extreme value of the image after the second moving operation is smaller than a negative preset local extreme value threshold value or not;
if so, determining that the moving direction of the third moving operation is opposite to the moving direction of the second moving operation;
if not, determining that the moving direction of the third moving operation is the same as the moving direction of the second moving operation.
Optionally, the step of determining whether to change the amplitude of the moving operation to a third preset amplitude smaller than the second preset amplitude according to the sharpness of the image before and after the third moving operation and a preset local extremum threshold includes:
obtaining a local extreme value of the image after the third moving operation through a preset formula according to the definition of the image before and after the third moving operation;
judging whether the local extreme value of the image after the third moving operation is smaller than a negative preset local extreme value threshold value or not;
if so, determining to change the amplitude of the moving operation to a third preset amplitude;
if not, determining not to change the amplitude of the moving operation to a third preset amplitude.
Optionally, the step of determining a moving direction of the third moving operation for the next time according to the definition of the image before and after the third moving operation, the preset local extremum threshold, and the accumulated number of consecutive moving times that includes the third moving operation and is the same as the moving direction of the third moving operation includes:
judging whether the local extreme value of the image after the third moving operation is larger than a preset local extreme value threshold value or not;
if so, determining that the moving direction of the next third moving operation is the same as the moving direction of the current third moving operation;
if not, judging whether the accumulated continuous moving times reach a second preset time or not;
if the accumulated continuous moving times reach a second preset time, determining that the moving direction of the third moving operation of the next time is opposite to the moving direction of the third moving operation of the current time;
and if the accumulated continuous moving times do not reach the second preset times, determining that the moving direction of the third moving operation of the next time is the same as the moving direction of the third moving operation of the current time.
Optionally, the preset formula is:
en ═ pn-pn-1)/min (pn, pn-1), where e isnFor local extrema, p, of the image after the shift operationnFor shifting the sharpness of the image after operation, pn-1To shift the sharpness of the image before the operation.
Optionally, the step of continuously performing the fourth moving operation for a first preset number of times in a third preset amplitude and in a direction opposite to the last third moving operation, and comparing the image after the fourth moving operation performed each time according to the third preset amplitude with the image after the last third moving operation to obtain the focused image includes:
taking the image after the last third moving operation as a candidate image;
executing a fourth movement operation for one time in a third preset amplitude and the opposite movement direction of the last third movement operation, and judging whether the image definition after the fourth movement operation for the time is greater than or equal to the candidate image definition;
if the image definition after the fourth movement operation is smaller than the candidate image definition, judging whether the cumulative times of the fourth movement operation executed according to a third preset amplitude reach a first preset time;
if the cumulative times of the fourth moving operation executed according to the third preset amplitude do not reach the first preset times, returning to execute the step of executing the fourth moving operation once in the third preset amplitude and the opposite moving direction of the last third moving operation;
if the accumulated times of the fourth moving operation executed according to the third preset amplitude reach the first preset times, taking the candidate image as a focused image;
if the image definition after the fourth moving operation is greater than or equal to the candidate image definition, changing the candidate image into the image after the fourth moving operation, and executing the following steps: and judging whether the cumulative number of times of the fourth moving operation executed according to the third preset amplitude reaches the first preset number of times.
Optionally, the step of comparing the image after the fourth moving operation performed at a third preset amplitude each time with the image after the last third moving operation to obtain the focused image includes:
and selecting the image with the maximum definition as the focused image from the image after the last third moving operation and the image after the fourth moving operation which is executed according to the third preset amplitude.
To achieve the above object, the present invention also provides a blood cell analyzer including a memory, a processor, and a computer program stored on the memory and executable on the processor, the computer program, when executed by the processor, implementing the steps of the auto-focusing method as described above.
To achieve the above object, the present invention also provides a computer-readable storage medium having stored thereon a computer program which, when being executed by a processor, realizes the steps of the auto-focusing method as described above.
The invention provides an automatic focusing method, a blood cell analyzer and a computer readable storage medium, which start reset according to a starting instruction input by a user and execute a first movement operation of a first preset amplitude for moving an objective table downwards; acquiring the definition of images before and after the movement operation with the first preset amplitude for the first time to determine the movement direction of the movement operation with the first preset amplitude for the second time, and executing the movement operation with the first preset amplitude for the second time according to the determined movement direction to acquire the definition of the images before and after the movement operation with the first preset amplitude for the second time; determining the moving direction of the moving operation with the second preset amplitude according to the definition of the image before and after the moving operation with the second first preset amplitude, the moving direction of the moving operation with the second first preset amplitude and the preset local extreme value threshold; executing a moving operation with a second preset amplitude according to the determined moving direction, and determining whether to change the amplitude of the moving operation to a third preset amplitude smaller than the second preset amplitude according to the definition of images before and after the moving operation with the second preset amplitude and a preset local extremum threshold; if not, determining the moving direction of the moving operation with the next second preset amplitude according to the definition of the images before and after the moving operation with the second preset amplitude, the preset local extremum threshold value and the accumulated continuous moving times which comprise the moving operation and are the same as the moving direction of the moving operation, returning to execute the moving operation with the second preset amplitude according to the determined moving direction, and determining whether to change the amplitude of the moving operation into a third preset amplitude smaller than the second preset amplitude according to the definition of the images before and after the moving operation with the second preset amplitude and the preset local extremum threshold value; if so, continuously executing the moving operation with the third preset amplitude for the first preset times in the opposite moving direction of the moving operation with the last second preset amplitude, and comparing the image after executing the moving operation with the third preset amplitude each time with the image after executing the moving operation with the last second preset amplitude to obtain the focused image. Due to the fact that the surface of the sample plate is uneven and noise factors, a plurality of false focal planes exist, and the local extreme value of the image in the false focal plane is different from the local extreme value of the image in the real focal plane, so that in the scheme, the blood cell analyzer moves the objective table to the real focal plane through the definition of the image before and after movement and the threshold value of the preset local extreme value in the process of moving the objective table by adopting the second preset amplitude, and then continuously moves the objective table by adopting the third preset amplitude to search the extreme value in the real focal plane. Therefore, the situation that a focal plane is searched only by comparing the image definition before and after movement in the process of using a traditional climbing method, the focal plane is easy to sink into a trap with a local extreme value, and a real focal plane is not easy to find is avoided.
Drawings
FIG. 1 is a schematic diagram of a hardware operating environment according to an embodiment of the present invention;
FIG. 2 is a flowchart illustrating a first embodiment of an auto-focusing method according to the present invention;
fig. 3 is a flowchart illustrating a detailed process of the step of determining the moving direction of the third moving operation performed at the second preset amplitude according to the sharpness of the image before and after the second moving operation, the moving direction of the second moving operation, and the preset local extremum threshold in step S30 in the second embodiment of the auto-focusing method according to the present invention;
fig. 4 is a flowchart illustrating a detailed process of the step of determining whether to change the amplitude of the moving operation to a third preset amplitude smaller than the second preset amplitude according to the sharpness of the image before and after the third moving operation and the preset local extremum threshold in step S30 in the third embodiment of the auto-focusing method according to the present invention;
FIG. 5 is a flowchart illustrating a detailed process of step S40 in the fourth embodiment of the auto-focusing method of the present invention;
fig. 6 is a detailed flowchart of step S50 in the fifth embodiment of the auto-focusing method according to the present invention.
The implementation, functional features and advantages of the objects of the present invention will be further explained with reference to the accompanying drawings.
Detailed Description
It should be understood that the specific embodiments described herein are merely illustrative of the invention and are not intended to limit the invention.
Referring to fig. 1, fig. 1 is a schematic diagram of a hardware structure of a blood cell analyzer according to various embodiments of the present invention. The blood cell analyzer includes a communication module 10, a memory 20, a processor 30, and the like. Those skilled in the art will appreciate that the blood cell analyzer shown in FIG. 1 may also include more or fewer components than shown, or combine certain components, or a different arrangement of components. Wherein, the processor 30 is connected to the memory 20 and the communication module 10, respectively, and the memory 20 stores thereon a computer program, which is executed by the processor 30 at the same time.
The communication module 10 may be connected to an external device through a network. The communication module 10 may receive data sent by an external device, and may also send data, instructions, and information to the external device, where the external device may be an electronic device such as a tablet computer, a notebook computer, and a desktop computer.
The memory 20 may be used to store software programs as well as various data. The memory 20 may mainly include a storage program area and a storage data area, wherein the storage program area may store an operating system, an application program (definition of acquired image) required for at least one function, and the like; the storage data area may store data or information created from use of the blood cell analyzer, or the like. Further, the memory 20 may include high speed random access memory, and may also include non-volatile memory, such as at least one magnetic disk storage device, flash memory device, or other volatile solid state storage device.
The processor 30, which is a control center of the blood cell analyzer, connects various parts of the whole blood cell analyzer by using various interfaces and lines, and performs various functions of the blood cell analyzer and processes data by operating or executing software programs and/or modules stored in the memory 20 and calling data stored in the memory 20, thereby performing overall monitoring of the blood cell analyzer. Processor 30 may include one or more processing units; preferably, the processor 30 may integrate an application processor, which mainly handles operating systems, user interfaces, application programs, etc., and a modem processor, which mainly handles wireless communications. It will be appreciated that the modem processor described above may not be integrated into the processor 30.
Although not shown in fig. 1, the blood cell analyzer may further include a circuit control module, which is used for being connected to a mains supply to implement power control and ensure normal operation of other components.
It will be understood by those skilled in the art that the configuration of the blood cell analyzer shown in FIG. 1 does not constitute a limitation of the blood cell analyzer and may include more or fewer components than shown, or some components may be combined, or a different arrangement of components.
Various embodiments of the method of the present invention are presented in terms of the above-described hardware architecture.
Referring to fig. 2, in a first embodiment of the auto-focusing method of the present invention, the auto-focusing method includes the steps of:
step S10, according to the start instruction input by the user, starting reset, and executing the first movement operation of moving the objective table downwards according to the first preset amplitude;
step S20, acquiring the sharpness of the image before and after the first moving operation to determine a moving direction of a second moving operation performed at a first preset magnitude, and performing the second moving operation according to the determined moving direction to acquire the sharpness of the image before and after the second moving operation;
in the present embodiment, the evaluation function of the image sharpness may be any one of gradient-based functions, such as Tenengrad function, energy gradient function, Brenner function, equation function, and the like.
The user places the sample at the objective table to after setting up the magnification, the user inputs and starts the automatic focusing instruction, after receiving the start automatic focusing instruction of user input, can reset the objective table to preset position at first, acquire the sample image that is in observing under the reset state, then carry out the first removal operation of moving the objective table downwards according to first predetermined amplitude, move the objective table downwards first predetermined amplitude automatically promptly. And then, acquiring a sample image observed after movement, determining a movement direction for executing second movement operation with a first preset amplitude by combining the definition of the sample image observed before movement, namely the definition of the sample image observed in a reset state and the definition of the sample image observed after movement, and finally acquiring the definition of the sample image observed before and after the second movement operation according to the determined movement direction and the first preset amplitude of the moving object stage after determining the direction of the second movement operation. If the determined moving direction is upward, the objective table is automatically moved upwards by a first preset amplitude, and if the determined moving direction is downward, the objective table is automatically moved downwards by the first preset amplitude.
The specific process of determining the moving direction for executing the second moving operation with the first preset amplitude is as follows:
step S21, judging whether the definition of the image after the first moving operation is larger than that of the image before the first moving operation; if yes, go to step S22, otherwise go to step S23;
step S22, determining the moving direction of the second moving operation to be downward;
in step S23, the moving direction of the second moving operation is determined to be upward.
Comparing whether the definition of the sample image observed after the first moving operation is greater than the definition of the sample image observed before the first moving operation (namely, the sample image observed in the reset state), namely, the definition of the sample image observed in the reset state, if so, indicating that the downward moving direction corresponding to the first moving operation is possibly correct, determining that the moving direction of the second moving operation continues to be downward movement; if the direction of the downward adjustment corresponding to the first moving operation is not larger than the preset value, the direction of the downward adjustment corresponding to the first moving operation is possibly wrong, and the moving direction of the second moving operation is determined to be changed into the upward movement.
Step S30, determining a moving direction of a third moving operation performed with a second preset amplitude according to the sharpness of the image before and after the second moving operation, the moving direction of the second moving operation, and a preset local extremum threshold, and performing the third moving operation according to the moving direction, and determining whether to change the amplitude of the moving operation to a third preset amplitude smaller than the second preset amplitude according to the sharpness of the image before and after the third moving operation and the preset local extremum threshold; if not, go to step S40; if yes, go to step S50;
the blood cell analyzer determines a moving direction of a third moving operation performed at a second preset magnitude based on the sharpness of the sample image observed after the second moving operation, the sharpness of the sample image observed before the second moving operation (i.e., the sample image observed after the first moving operation), the moving direction of the second moving operation, and a preset local extremum threshold.
After the moving direction of the third moving operation performed with the second preset amplitude is determined, the blood cell analyzer may perform the third moving operation according to the determined moving direction, that is, automatically move the stage by the second preset amplitude according to the determined moving direction, then determine whether to change the moving amplitude of the moving operation to the third preset amplitude smaller than the second preset amplitude according to the sharpness of the sample image observed before the third moving operation and the sharpness of the sample image observed after the third moving operation, and the preset local extremum threshold, and if it is determined that the moving amplitude is not changed to the third preset amplitude, that is, it is determined that the stage is continuously performed with the second preset amplitude, then perform step S40; if it is determined that the moving amplitude is changed to the third preset amplitude, that is, the fourth moving operation is performed to move the stage using the third preset amplitude, step S50 is performed.
It should be noted that the first preset amplitude may be greater than or equal to or smaller than the second preset amplitude, but it is generally preferable that the second preset amplitude is greater than or equal to the first preset amplitude.
Step S40, determining the moving direction of the third moving operation in the next time according to the definition of the image before and after the third moving operation, the preset local extremum threshold and the accumulated continuous moving times which comprise the third moving operation and have the same moving direction with the third moving operation, and returning to the step of executing the third moving operation according to the moving direction;
when the blood cell analyzer determines not to change the moving amplitude to the third preset amplitude, namely determines to continue to adopt the second preset amplitude to move the objective table, determining the moving direction of the next third moving operation according to the definition of the sample image observed before and after the third moving operation, the preset local extremum threshold and the accumulated continuous moving times including the third moving operation and having the same moving direction as the third moving operation (if the accumulated continuous moving times including the third moving operation and having the same moving direction as the third moving operation is 4 times, it indicates that the moving direction of the third moving operation corresponding to the 4 th movement pushed forward from the third moving operation is opposite to the moving direction of the third moving operation or the moving operation corresponding to the 4 th movement pushed forward from the third moving operation is the second moving operation), and returning to continue to execute the third moving operation according to the moving direction.
Step S50, continuously executing a fourth moving operation for a first preset number of times in a third preset amplitude and in a direction opposite to the last third moving operation, and comparing the image after the fourth moving operation executed each time according to the third preset amplitude with the image after the last third moving operation to obtain a focused image.
When the blood cell analyzer determines to change the moving amplitude to a third preset amplitude, namely, after the object stage is determined to be moved by the third preset amplitude subsequently, continuously executing a fourth moving operation for a first preset number of times in the opposite moving direction to the last third moving operation and in the third preset amplitude, namely, continuously moving the object stage for a third preset number of times in the opposite moving direction to the last third moving operation, wherein the continuous moving number of times is the first preset number of times, acquiring a sample image observed after the fourth moving operation executed according to the third preset amplitude each time, and then comparing the sample image observed after the fourth moving operation executed according to the third preset amplitude each time with the image after the third moving operation at the last time to obtain a focused image.
It should be noted that after determining to change the movement amplitude to the third preset amplitude smaller than the second preset amplitude, step S50 may not be executed to obtain the focused image, and step S50 may be executedThe focused image is obtained through the following steps: after the moving amplitude is determined to be changed from the second preset amplitude to the third preset amplitude, continuously executing the fourth moving operation for the first preset times in the opposite moving direction of the third preset amplitude and the last third moving operation, after the third preset amplitude of the object stage is moved each time, not only obtaining the sample image observed after each moving, but also obtaining the position of the object stage after each moving, determining the image with the maximum definition according to the image after the fourth moving operation executed according to the third preset amplitude each time and the image after the last third moving operation, and moving the object stage to the position w when the obtained image with the maximum definition is moved0And within a third predetermined range of amplitudes from and above the position, i.e. (w)0-d3，w0+d3) Obtaining focused images within a range by adopting a hill climbing method, wherein d3Is a third predetermined magnitude. In (w)0-d3，w0+d3) The specific process of obtaining the focused image by adopting the hill climbing method in the range comprises the following steps: first, at position w0Is an initial position, and acquires a position w0The clarity of the observed sample image F (w)0) Then selecting the upward or downward direction as the moving direction, and setting the moving direction to be a fourth preset amplitude d4Moving objective table (d)4≤d3/2), the arrival position w1 ═ w0-d4(moving direction upward) or w0+d4(moving direction is downward), and acquiring the definition F (w) of the observed sample image after the movement1) Judgment of F (w)1) Whether or not it is greater than F (w)0) If F (w)1) Greater than F (w)0) Then the direction is kept unchanged and the stage is moved by a fourth preset amplitude until F (w)n-1) Greater than F (w)n) Then, the movement is reversed, and the movement amplitude is changed to be half or less of the movement amplitude in the non-reversed forward movement process, and the movement process is the same as the forward movement process, and the movement is repeated until the definition F (w) of the sample image is observed before the movementi) And the sharpness F (w) of the observed sample image after shiftingi+1) The absolute value of the difference between the two is less than or equal to a preset threshold value, at the moment, the bit is determinedPut wi+1For a focus position, the image of the sample observed at that position is taken as a focused image. By this process, the focus position can be found more accurately.
The embodiment starts reset according to a starting instruction input by a user and executes a first moving operation of moving the objective table downwards according to a first preset amplitude; acquiring the definition of images before and after the first moving operation to determine the moving direction of a second moving operation executed in a first preset amplitude, and executing the second moving operation according to the determined moving direction to acquire the definition of the images before and after the second moving operation; determining the moving direction of a third moving operation executed in a second preset amplitude according to the definition of the images before and after the second moving operation, the moving direction of the second moving operation and a preset local extremum threshold, executing the third moving operation according to the moving direction, and determining whether to change the amplitude of the moving operation to a third preset amplitude smaller than the second preset amplitude according to the definition of the images before and after the third moving operation and the preset local extremum threshold; if not, determining the moving direction of the next third moving operation according to the definition of the image before and after the third moving operation, a preset local extremum threshold and the accumulated continuous moving times which comprise the third moving operation and have the same moving direction as the third moving operation, and returning to the step of executing the third moving operation according to the moving direction; if so, continuously executing the fourth moving operation for a first preset number of times in a third preset amplitude and the opposite moving direction of the last third moving operation, and comparing the image after the fourth moving operation executed according to the third preset amplitude each time with the image after the last third moving operation to obtain the focused image. Due to the fact that the surface of the sample plate is uneven and noise factors, a plurality of false focal planes exist, and the local extreme value of the image in the false focal plane is different from the local extreme value of the image in the real focal plane, so that in the scheme, the blood cell analyzer moves the objective table to the real focal plane through the definition of the image before and after movement and the threshold value of the preset local extreme value in the process of moving the objective table by adopting the second preset amplitude, and then continuously moves the objective table by adopting the third preset amplitude to search the extreme value in the real focal plane. Therefore, the situation that a focal plane is searched only by comparing the image definition before and after movement in the process of using a traditional climbing method, the focal plane is easy to sink into a trap with a local extreme value, and a real focal plane is not easy to find is avoided.
Further, referring to fig. 3, a second embodiment of the auto-focusing method of the present application is proposed according to the first embodiment of the auto-focusing method of the present application, and in this embodiment, the step of determining the moving direction of the third moving operation performed at the second preset magnitude according to the sharpness of the image before and after the second moving operation, the moving direction of the second moving operation, and the preset local extremum threshold in step S30 includes:
step S31, obtaining a local extreme value of the image after the second moving operation through a preset formula according to the definition of the image before and after the second moving operation;
step S32, judging whether the local extreme value of the image after the second moving operation is smaller than a negative preset local extreme value threshold value; if yes, go to step S33; if not, go to step S34;
step S33, determining that the moving direction of the third moving operation is opposite to the moving direction of the second moving operation;
in step S34, it is determined that the moving direction of the third moving operation is the same as the moving direction of the second moving operation.
In this example, after acquiring the sample images observed before and after the second shift operation, the blood cell analyzer inputs the sharpness of the sample image observed before the second shift operation and the sharpness of the sample image observed after the second shift operation into a preset formula en ═ pn-1/min (pn, pn-1) to obtain a local extreme value of the sample image observed after the second shift operation, where e isnFor local extrema, p, of the image after the shift operationnFor shifting the sharpness of the image after operation, pn-1To shift the sharpness of the image before the operation.
After the blood cell analyzer acquires the local extreme value of the sample image observed after the second moving operation, whether the local extreme value of the sample image observed after the second moving operation is smaller than a negative preset local extreme value threshold value or not is judged, if the local extreme value of the sample image observed after the second moving operation is smaller than the negative preset local extreme value threshold value is determined, the blood cell analyzer determines that the moving direction of the next third moving operation is opposite to the moving direction of the second moving operation, for example, the moving direction of the second moving operation is upward, and the moving direction of the next third moving operation is downward; if the local extreme value of the sample image observed after the second moving operation is determined to be not less than the negative preset local extreme value threshold, the blood cell analyzer determines that the moving direction of the next third moving operation is the same as the moving direction of the second moving operation, for example, the moving direction of the second moving operation is upward, and the moving direction of the next third moving operation is still upward.
In the embodiment, the initial moving direction in the process of searching the real focal plane by using the moving object stage with the second preset amplitude is determined by using the definition and the local extremum threshold before and after the moving object stage with the first preset amplitude for the last time.
Further, referring to fig. 4, a third embodiment of the auto-focusing method according to the first embodiment of the auto-focusing method is provided, in which the step of determining whether to change the amplitude of the moving operation to a third preset amplitude smaller than the second preset amplitude according to the sharpness of the image before and after the third moving operation and the preset local extremum threshold in step S30 includes:
step S35, obtaining a local extreme value of the image after the third moving operation through a preset formula according to the definition of the image before and after the third moving operation;
step S36, judging whether the local extreme value of the image after the third moving operation is smaller than a negative preset local extreme value threshold value; if yes, go to step S37; if not, go to step S38;
step S37, determining to change the amplitude of the moving operation to a third preset amplitude;
in step S38, it is determined not to change the magnitude of the moving operation to the third preset magnitude.
In this example, the hematology analyzer acquires the sample observed before and after the third movement operationAnd inputting the definition of the sample image observed before the third moving operation and the definition of the sample image observed after the third moving operation into a preset formula en ═ pn-pn-1/min (pn, pn-1) so as to obtain a local extreme value of the sample image observed after the third moving operation, wherein e isnFor local extrema, p, of the image after the shift operationnFor shifting the sharpness of the image after operation, pn-1To shift the sharpness of the image before the operation.
After the blood cell analyzer obtains the local extreme value of the sample image observed after the third moving operation, whether the local extreme value of the sample image observed after the third moving operation is smaller than a negative preset local extreme value threshold value or not is judged, and when the local extreme value of the sample image observed after the third moving operation is determined to be smaller than the negative preset local extreme value threshold value, the blood cell analyzer determines to change the moving amplitude from the second preset amplitude to a third preset amplitude smaller than the second preset amplitude, namely, the blood cell analyzer enters a small-amplitude moving mode; and when the local extreme value of the sample image observed after the third movement operation is determined to be not less than the negative preset local extreme value threshold value, the blood cell analyzer continuously adopts the second preset amplitude to move the object stage, namely, the large-amplitude movement mode is continuously adopted.
The embodiment provides a strategy for judging whether to find a real focal plane or not by comparing the actual local extreme value of an observed sample image after movement with the preset local extreme value threshold value after the object stage is moved by adopting a large-amplitude moving mode, so that the situation that the focal plane is searched by only comparing the image definition before and after movement in the process of using a traditional climbing method and is easily trapped in a trap of the local extreme value, and the real focal plane is not easily found is avoided.
Further, referring to fig. 5, a fourth embodiment of the auto-focusing method of the present application is proposed according to the first embodiment of the auto-focusing method of the present application, and in this embodiment, step S40 includes:
step S41, judging whether the local extreme value of the image after the third moving operation is larger than a preset local extreme value threshold value; if yes, go to step S42; if not, go to step S43;
step S42, determining that the moving direction of the next third moving operation is the same as the moving direction of the present third moving operation;
step S43, judging whether the accumulated continuous moving times reaches a second preset time; if yes, go to step S44; if not, go to step S45;
step S44, determining that the moving direction of the next third moving operation is opposite to the moving direction of the present third moving operation;
in step S45, it is determined that the moving direction of the next third moving operation is the same as the moving direction of the present third moving operation.
In this example, after obtaining the local extreme value of the sample image observed after the third moving operation, the hematology analyzer may determine whether the local extreme value of the sample image observed after the third moving operation is greater than a preset local extreme value threshold, and when determining that the local extreme value of the sample image observed after the third moving operation is greater than the preset local extreme value threshold, the hematology analyzer determines that the moving direction of the next third moving operation is the same as the moving direction of the third moving operation, for example, the moving direction of the third moving operation is upward, the moving direction of the next third moving operation is still upward; when the local extreme value of the sample image observed after the third movement operation is determined to be not greater than the preset local extreme value threshold value, continuously judging whether the accumulated continuous movement times which comprise the third movement operation and are the same as the movement direction of the third movement operation reach a second preset time, and when the accumulated continuous movement times reach the second preset time, determining that the movement direction of the next third movement operation is opposite to the movement direction of the current third movement operation by the blood cell analyzer, for example, the movement direction of the current third movement reference is upward, and the movement direction of the next third movement operation is downward; when the second preset number of times is not reached, the blood cell analyzer determines that the moving direction of the next third moving operation is the same as the moving direction of the current third moving operation, for example, the moving direction of the current third moving operation is upward, and the moving direction of the next third moving operation is still upward.
The embodiment provides a moving strategy for the next step, which is formulated according to the local extreme value of the sample image after moving and the continuous moving times which are the same as the current moving direction when the local extreme value of the sample image observed after moving does not meet the local extreme value range required by the real focal plane in the process of searching the real focal plane by adopting the second preset amplitude moving object stage.
Further, referring to fig. 6, a fifth embodiment of the auto-focusing method of the present application is proposed according to the first embodiment of the auto-focusing method of the present application, and in this embodiment, step S50 includes:
step S51, taking the image after the last third moving operation as a candidate image;
step S52, executing a fourth movement operation for one time in a third preset amplitude and the opposite movement direction of the last third movement operation, and judging whether the image definition after the fourth movement operation for the time is more than or equal to the candidate image definition; if not, go to step S53; if yes, go to step S54;
step S53, determining whether the cumulative number of fourth move operations performed according to the third preset magnitude reaches the first preset number; if not, returning to the step S52; if yes, go to step S55;
step S54, changing the candidate image into the image after the fourth movement operation, and returning to execute step S53;
in step S55, the candidate image is set as the focused image.
In this example, the blood cell analyzer uses the last third movement operation, that is, the last sample image observed by using the second preset stage as a candidate image, then moves the stage for the third preset amplitude in the opposite movement direction to the last third movement operation, and then determines whether the sample image observed after the fourth movement operation is greater than or equal to the candidate image definition, if the sample image observed after the fourth movement operation is not greater than or equal to the candidate image definition, then determines whether the cumulative number of times of the fourth movement operation performed according to the third preset amplitude reaches the first preset number of times, and when the cumulative number of times of the fourth movement operation performed by the blood cell analyzer according to the third preset amplitude does not reach the first preset number of times, then continuously returns to perform the step of performing the fourth movement operation for one time in the opposite movement direction to the third preset amplitude and the last third movement operation, when the cumulative number of fourth moving operations executed by the blood cell analyzer according to the third preset amplitude reaches the first preset number, taking the candidate image as a focused image; and if the observed sample image after the fourth moving operation is greater than or equal to the definition of the candidate image, changing the candidate image into the observed sample image after the fourth moving operation, and returning to the step of judging whether the cumulative number of the fourth moving operation executed according to the third preset amplitude reaches the first preset number.
For example, the first preset number of times is 3, the blood cell analyzer firstly uses the sample image P0 observed after the stage is moved for the last time by the second preset amplitude as a candidate image, then obtains the sample image P1 observed at this time after the stage is moved for the first time by the third preset amplitude, compares the definition of the sample image P1 with the definition of the sample image P0, if so, changes the candidate image to P1, and judges whether the cumulative number of times of movement by the third preset amplitude reaches 3 times, if not, the candidate image is still P0, and directly judges whether the cumulative number of times of movement by the third preset amplitude reaches 3 times. At this time, the accumulated movement times with the third preset amplitude is 1, and the movement times do not reach 3 times, the third preset amplitude is continuously adopted to carry out the second movement of the object stage, the observed sample image after movement is obtained as P2, whether the definition of the sample image P2 is greater than that of the candidate image or not is continuously judged, if the definition of P2 is greater than that of the candidate image, the candidate image is changed into P2, and if the definition of P2 is not greater than that of the candidate image, the candidate image is not changed. And continuously judging whether the accumulated moving times with the third preset amplitude reach 3 times or not. At this time, the accumulated moving times with the third preset amplitude is 2, and the third preset amplitude is not used for 3 times, the objective table is continuously moved for the third time, the observed sample image after moving is P3, whether the definition of the sample image P3 is larger than the definition of the candidate image or not is continuously judged, if the definition of P3 is larger than the definition of the candidate image, the candidate image is changed to P3, and if the definition of P3 is not larger than the definition of the candidate image, the candidate image is not changed, and the accumulated moving times with the third preset amplitude is 3 at this time, and the candidate image reaches 3 times, the candidate image is used as the focused image.
Due to the fact that the surface of the sample plate is uneven and noisy, besides a plurality of false focal planes, a plurality of false extreme values can exist in the focal plane.
Further, a sixth embodiment of the auto-focusing method according to the present application is proposed according to the first embodiment of the auto-focusing method, and in this embodiment, the step of comparing the image after the fourth moving operation performed at each time by the third preset magnitude with the image after the last third moving operation in step S50 to obtain the focused image includes:
in step S56, an image with the greatest sharpness is selected as a focused image from the image after the last third shift operation and the images after the fourth shift operations each performed at the third preset magnitude.
In this example, after the blood cell analyzer continuously performs the fourth moving operation for the first preset number of times with the third preset width, an image with the maximum sharpness is selected as a focused image from among the observed sample images after the last third moving operation and the observed sample images after the fourth moving operation performed at each of the third preset widths. For example, the first preset number of times is 3, the last time the stage is moved by the second preset amplitude, the observed sample image P0 is observed, the image after the stage is moved by the third preset amplitude for the first time is P1, the image after the stage is moved by the third preset amplitude for the second time is P2, and the image after the stage is moved by the third preset amplitude for the third time is P3, wherein the image P2 has the maximum definition, and the image with the maximum definition P2 is selected as the focused image.
Due to the fact that the surface of the sample plate is uneven and noise factors, besides a plurality of false focal planes, a plurality of false extreme values can exist in the focal plane, the third preset amplitude is adopted to move in the focal plane for a plurality of times, the definition of the image after each movement is compared together, the true extreme value position, namely the true focusing position, in the focal plane is found, and compared with the previous embodiment, the process of comparing the image definition before and after each movement is adopted to obtain the true focusing position, and the process of the example is simpler.
The invention also proposes a computer-readable storage medium on which a computer program is stored. The computer readable storage medium may be the Memory 20 of the blood cell analyzer of fig. 1, and may be at least one of a ROM (Read-Only Memory)/RAM (Random Access Memory), a magnetic disk, and an optical disk, and the computer readable storage medium includes information for causing the blood cell analyzer to perform the method according to the embodiments of the present invention.
It should be noted that, in this document, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or system that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or system. Without further limitation, an element defined by the phrase "comprising an … …" does not exclude the presence of other like elements in a process, method, article, or system that comprises the element.
The above-mentioned serial numbers of the embodiments of the present invention are merely for description and do not represent the merits of the embodiments.
Through the above description of the embodiments, those skilled in the art will clearly understand that the method of the above embodiments can be implemented by software plus a necessary general hardware platform, and certainly can also be implemented by hardware, but in many cases, the former is a better implementation manner.
The above description is only a preferred embodiment of the present invention, and not intended to limit the scope of the present invention, and all modifications of equivalent structures and equivalent processes, which are made by using the contents of the present specification and the accompanying drawings, or directly or indirectly applied to other related technical fields, are included in the scope of the present invention.
Claims (8)
1. An auto-focusing method, comprising the steps of:
starting reset according to a starting instruction input by a user, and executing a first moving operation of moving the objective table downwards according to a first preset amplitude;
acquiring the definition of images before and after the first moving operation to determine the moving direction of a second moving operation executed in a first preset amplitude, and executing the second moving operation according to the determined moving direction to acquire the definition of the images before and after the second moving operation;
determining the moving direction of a third moving operation executed in a second preset amplitude according to the definition of the images before and after the second moving operation, the moving direction of the second moving operation and a preset local extremum threshold, executing the third moving operation according to the moving direction, and determining whether to change the amplitude of the moving operation to a third preset amplitude smaller than the second preset amplitude according to the definition of the images before and after the third moving operation and the preset local extremum threshold;
if not, determining the moving direction of the next third moving operation according to the definition of the image before and after the third moving operation, a preset local extremum threshold and the accumulated continuous moving times which comprise the third moving operation and have the same moving direction as the third moving operation, and returning to the step of executing the third moving operation according to the moving direction;
if so, continuously executing a fourth moving operation for a first preset number of times in a third preset amplitude and the opposite moving direction of the last third moving operation, and comparing an image after the fourth moving operation executed according to the third preset amplitude each time with an image after the last third moving operation to obtain a focused image;
wherein, the step of determining the moving direction of the third moving operation executed at the second preset amplitude according to the definition of the image before and after the second moving operation, the moving direction of the second moving operation and the preset local extremum threshold comprises:
obtaining a local extreme value of the image after the second moving operation through a preset formula according to the definition of the image before and after the second moving operation;
judging whether the local extreme value of the image after the second moving operation is smaller than a negative preset local extreme value threshold value or not;
if so, determining that the moving direction of the third moving operation is opposite to the moving direction of the second moving operation;
if not, determining that the moving direction of the third moving operation is the same as the moving direction of the second moving operation;
the preset formula is as follows: e.g. of the typen＝(pn-pn-1)/min(pn,pn-1)，
Wherein enFor local extrema, p, of the image after the shift operationnFor shifting the sharpness of the image after operation, pn-1To shift the sharpness of the image before the operation.
2. The auto-focusing method of claim 1, wherein the step of acquiring the sharpness of the image before and after the first moving operation to determine the moving direction of the second moving operation performed at the first preset magnitude comprises:
judging whether the definition of the image after the first moving operation is greater than that of the image before the first moving operation;
if so, determining that the moving direction of the second moving operation is downward;
if not, determining that the moving direction of the second moving operation is upward.
3. The auto-focusing method of claim 2, wherein the step of determining whether to change the amplitude of the moving operation to a third preset amplitude smaller than the second preset amplitude according to the sharpness of the image before and after the third moving operation and the preset local extremum threshold comprises:
obtaining a local extreme value of the image after the third moving operation through a preset formula according to the definition of the image before and after the third moving operation;
judging whether the local extreme value of the image after the third moving operation is smaller than a negative preset local extreme value threshold value or not;
if so, determining to change the amplitude of the moving operation to a third preset amplitude;
if not, determining not to change the amplitude of the moving operation to a third preset amplitude.
4. The auto-focusing method of claim 3, wherein the step of determining the moving direction of the next third moving operation according to the sharpness of the image before and after the third moving operation, the preset local extremum threshold, and the accumulated number of consecutive moving times including the third moving operation and being the same as the moving direction of the third moving operation comprises:
judging whether the local extreme value of the image after the third moving operation is larger than a preset local extreme value threshold value or not;
if so, determining that the moving direction of the next third moving operation is the same as the moving direction of the current third moving operation;
if not, judging whether the accumulated continuous moving times reach a second preset time or not;
if the accumulated continuous moving times reach a second preset time, determining that the moving direction of the third moving operation of the next time is opposite to the moving direction of the third moving operation of the current time;
and if the accumulated continuous moving times do not reach the second preset times, determining that the moving direction of the third moving operation of the next time is the same as the moving direction of the third moving operation of the current time.
5. The auto-focusing method of any one of claims 1 to 4, wherein the step of consecutively performing the fourth moving operation for a first preset number of times in a third preset magnitude and in a direction opposite to the last third moving operation, and comparing the image after the fourth moving operation performed each time at the third preset magnitude with the image after the last third moving operation to obtain the focused image comprises:
taking the image after the last third moving operation as a candidate image;
executing a fourth movement operation for one time in a third preset amplitude and the opposite movement direction of the last third movement operation, and judging whether the image definition after the fourth movement operation for the time is greater than or equal to the candidate image definition;
if the image definition after the fourth movement operation is smaller than the candidate image definition, judging whether the cumulative times of the fourth movement operation executed according to a third preset amplitude reach a first preset time;
if the cumulative times of the fourth moving operation executed according to the third preset amplitude do not reach the first preset times, returning to execute the step of executing the fourth moving operation once in the third preset amplitude and the opposite moving direction of the last third moving operation;
if the accumulated times of the fourth moving operation executed according to the third preset amplitude reach the first preset times, taking the candidate image as a focused image;
if the image definition after the fourth moving operation is greater than or equal to the candidate image definition, changing the candidate image into the image after the fourth moving operation, and executing the following steps: and judging whether the cumulative number of times of the fourth moving operation executed according to the third preset amplitude reaches the first preset number of times.
6. The auto-focusing method according to any one of claims 1 to 4, wherein the step of comparing the image after the fourth moving operation performed each time at a third preset magnitude with the image after the last third moving operation to obtain the focused image comprises:
and selecting the image with the maximum definition as the focused image from the image after the last third moving operation and the image after the fourth moving operation which is executed according to the third preset amplitude.
7. A blood cell analyzer, characterized in that it comprises a memory, a processor and a computer program stored on the memory and executable on the processor, which computer program, when executed by the processor, carries out the steps of the autofocus method according to any one of claims 1 to 6.
8. A computer-readable storage medium, characterized in that a computer program is stored thereon, which computer program, when being executed by a processor, carries out the steps of the autofocus method as claimed in one of claims 1 to 6.
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