CN115992183B - Sperm automatic braking and sucking method and system - Google Patents
Sperm automatic braking and sucking method and system Download PDFInfo
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- CN115992183B CN115992183B CN202310275839.5A CN202310275839A CN115992183B CN 115992183 B CN115992183 B CN 115992183B CN 202310275839 A CN202310275839 A CN 202310275839A CN 115992183 B CN115992183 B CN 115992183B
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- Y02A40/00—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production
- Y02A40/80—Adaptation technologies in agriculture, forestry, livestock or agroalimentary production in fisheries management
- Y02A40/81—Aquaculture, e.g. of fish
Abstract
The invention provides a sperm automatic braking and sucking method and a sperm automatic braking and sucking system. The invention provides a method for automatically braking and sucking sperms by operating a micropipette for the first time, and a controllable sperm liquid substrate workbench is not needed. Thanks to the sperm scoring mechanism and the motion control strategy of the invention, the invention can autonomously select or assist an operator to rapidly select sperm in the field of view suitable for operation; turbulence of the sperm cell process tool motion on the sperm cell containing liquid substrate can be avoided or compensated for. The invention simplifies the constitution of the sperm operation system, is more close to the current equipment conditions of most hospitals, and reduces the development cost of the sperm operation system.
Description
Technical Field
The invention relates to the technical field of microscopic cell operation, in particular to a sperm automatic braking and sucking method and system based on a single positioner.
Background
Intracytoplasmic single sperm injection (ICSI) is an important procedure in vitro fertilization, the main steps of which include sperm manipulation (sperm arrest and sperm aspiration) and cell injection. Traditionally, sperm manipulation is accomplished by manual manipulation, but the learning period of this skill is long and the success rate is limited by the skill level of the operator. Automation of sperm manipulation has been widely studied during the last decade, but current sperm manipulation techniques mostly rely on the combined manipulation of multiple positioners, making the sperm manipulation system costly and quite different from the instrumentation currently deployed in most hospitals. If the sperm cell manipulation system is simplified, only a single positioner is used to control the sperm cell manipulation tool, the problem may be significantly ameliorated.
The application of the existing sperm cell operation technology to the simplified sperm cell operation system has the following defects:
(1) The sperm selection process does not take into account the problem of sperm exiting the field of view of the image acquisition device. During operation, sperm remain in motion throughout the liquid substrate. Thus, sperm cells that are at the edge of the field of view or that are farther from the end of the handling tool are more likely to be out of view during handling, resulting in a failure of the handling.
(2) Most of the current sperm cell manipulation methods first adjust the sperm cell to be manipulated and the end of the manipulation tool to the same position on the image plane, and then perform a braking operation normal to the image plane. However, during the process of the handling tool tip tracking sperm, the handling tool tip moves rapidly in the liquid substrate containing sperm. Due to the viscosity of the liquid substrate, the movement of the tool tip causes disturbances to the substrate and sperm therein, causing them to produce undesired movements, resulting in sperm tracking errors.
Therefore, in view of the above problems, it is necessary to propose a further solution.
Disclosure of Invention
The invention aims to provide a sperm automatic braking and sucking method and system based on a single positioner, which overcome the defects in the prior art.
In order to achieve the above object, the present invention provides an automatic sperm braking and sucking method, comprising the steps of:
s1, placing a liquid substrate containing sperms on a workbench, enabling the liquid substrate to be located in an imaging range of an image data display unit, acquiring images of the tip of a sperm operation tool, sperms and the liquid substrate by using an image data acquisition unit, and transmitting the images to a host unit;
s2, the host unit controls the sperm operation tool through the control locator according to the distance between the sperm operation tool tip and the sperm in the image, so that the sperm operation tool tip tracks and moves the sperm in the direction parallel to the image plane until the sperm operation tool tip and the sperm are positioned at the same position in the image plane;
s3, using a positioner to enable the tip of the sperm operating tool and the mobile sperm to remain relatively static in an image plane and move normally in the image plane, and realizing sperm braking through physical interaction between the tip of the sperm operating tool and the mobile sperm;
s4, adjusting the pressure in the cavity of the sperm operation tool, and automatically sucking and positioning the braking sperm.
As an improvement of the method of the present invention, the step S1 further includes a step of automatically or actively selecting sperm to be handled by the system or by an operator.
As an improvement of the method, the method also comprises the step of generating each sperm fraction according to the expected movement track of each sperm, the orientation of each sperm and the distance between each sperm and the center of the visual field through visual feedback of the sperm and the liquid substrate so as to realize the automatic or auxiliary active selection of the sperm to be operated by a system operator.
As an improvement of the method of the present invention, the method further comprises:
updating the fraction of each sperm in real time according to the changes of the direction, the distance and the expected movement track caused by the movement of each sperm, and considering the switching cost of an operation target generated by the replacement of the sperm in the operation process, so as to replace the sperm to be operated when necessary;
the method further comprises the steps of:
the switching cost of the operation target is considered in the process of automatically selecting and replacing the sperm to be operated so as to avoid frequent replacement of the sperm to be operated.
As an improvement of the method of the present invention, said step S2 further comprises a step of tracking sperm at the image plane, comprising:
the open loop method comprises the following steps: firstly, determining the position of the sperm in an image plane according to image feedback of the tip of a sperm operation tool, then using a positioner to normally lift the tip of the sperm operation tool in the image plane, and controlling the position of the tip of the sperm operation tool according to a moving sperm real-time image in an open loop manner to realize sperm tracking; or:
the closed loop method comprises the following steps: and observing the positions of the tip and the sperm of the sperm manipulation tool in real time on a focal plane, performing closed-loop motion control, and compensating the passive motion process of the sperm to be manipulated caused by the disturbance of the liquid substrate by the sperm manipulation tool, so as to accelerate the tip of the sperm manipulation tool to track the sperm to be manipulated.
As an improvement of the method of the present invention, the step S3 further includes:
predicting and compensating the movement of the mobile sperm to reduce the relative error of tracking the mobile sperm, and stopping the sperm to be operated by moving the sperm cell manipulator tip normal to the image plane to contact the bottom of the liquid substrate and squeeze the mobile sperm.
To achieve the above object, the present invention provides a system for implementing the sperm automatic braking and sucking method as described above, comprising: the device comprises a workbench, an image data display unit, an image data acquisition unit, a micropipette unit and a host unit;
the workbench is arranged in the imaging range of the image data display unit;
the image data acquisition unit and the image data display unit perform image transmission through an optical path;
the micropipette unit comprises a locator connected to a sperm manipulation tool;
the operation end of the sperm cell operation tool extends to the upper part of the workbench;
the host unit is in data transmission with the image data acquisition unit, and processes the image data sent by the image data acquisition unit to obtain the distance between sperms in the image data and the operating end of the sperm operating tool, and the host unit controls the locator to drive the sperm operating tool to brake and absorb according to the distance.
As an improvement to the system of the present invention, the sperm manipulation tool is a micropipette having a hollow needle-like tip.
As an improvement of the system of the invention, the micropipette unit further comprises a pressure mechanism which is communicated with the sperm cell manipulation tool through a conduit, and which is controlled by the host unit to cause the sperm cell manipulation tool to perform a pipetting action.
As an improvement of the system of the invention, the pressure mechanism is a pressure pump.
Compared with the prior art, the invention has the beneficial effects that: the invention provides a method for automatically braking and sucking sperms by operating a micropipette for the first time, and a controllable sperm liquid substrate workbench is not needed. Thanks to the sperm scoring mechanism and the motion control strategy of the invention, the invention can autonomously select or assist an operator to rapidly select sperm in the field of view suitable for operation; turbulence of the sperm cell process tool motion on the sperm cell containing liquid substrate can be avoided or compensated for. The invention simplifies the constitution of the sperm operation system, is more close to the current equipment conditions of most hospitals, and reduces the development cost of the sperm operation system.
Drawings
In order to more clearly illustrate the embodiments of the present invention or the technical solutions in the prior art, the drawings that are required to be used in the embodiments or the description of the prior art will be briefly described below, and it is obvious that the drawings in the following description are only some embodiments described in the present invention, and other drawings may be obtained according to the drawings without inventive effort to those skilled in the art.
FIG. 1 is a schematic flow chart of a method for automatically braking and sucking sperm according to an embodiment of the present invention;
FIG. 2 is a diagram of a sperm scoring method in accordance with an embodiment of the present invention;
fig. 3 is a schematic structural diagram of an automatic sperm braking and sucking system according to an embodiment of the present invention.
Detailed Description
The present invention will be described in detail with reference to the following embodiments, but it should be understood that these embodiments are not limiting, and functional, method, or structural equivalents and alternatives thereof by those skilled in the art are within the scope of the present invention.
As shown in fig. 1, the present embodiment provides a method for achieving automatic sperm braking and aspiration using a single positioner. The automatic sperm braking and sucking method of the embodiment specifically comprises the following steps:
s1, placing a liquid substrate containing sperms in a microscopic visible range on a workbench, and acquiring images of the tip of a sperm operation tool, the sperms and the liquid substrate by using an image data acquisition unit and transmitting the images to a host unit.
In the implementation process, the sperm and liquid substrate images acquired by the image data acquisition unit are tracked by adopting a discrimination related filtering method, and parameters such as the position, the kinematics and the morphology of the sperm are calculated.
In the process of tracking sperms, the sperms suitable for operation need to be selected manually or automatically, so the invention provides a method for scoring each sperm in a screen, wherein the scoring standard comprises the motion track of each sperm, the distance between each sperm and the center of the visual field and the tip of the micropipette, and the orientation of each sperm, and the scoring mode is shown in figure 2. Wherein the expected motion trajectories of each sperm are calculated based on an extended kalman filter method. The total score of each sperm is calculated by the weighted average of the sub scores of the four indexes, and the sub scores of the indexes are respectively nonlinear relations of the corresponding parameters. The scoring method comprises the following steps:
wherein, the liquid crystal display device comprises a liquid crystal display device,cfor the comprehensive scoring of the sperm to be handled,c 1 …c 4 a kind of electronic device with high-pressure air-conditioning systemw 1 …w 4 The individual scores and weights of the four indexes of the sperm to be operated are respectively,
being a positive constant, abs (·) represents absolute value, E (·) represents desired, z i The first sperm to be handlediPredicted position of step->
Represent the firstiLikelihood of step>
The center of the field of view, the location of the sperm and tip,k 1 , k 21 , k 22 , k 3 , k 4 is the gain.
S2, the host unit controls the sperm operating tool through the control locator according to the distance between the sperm operating tool tip and the sperm in the image, so that the sperm operating tool tip tracks and moves the sperm in the direction parallel to the image plane until the sperm operating tool tip and the sperm are positioned at the same position in the image plane.
In the implementation process, firstly, the sperm to be operated needs to be selected. In the process of tracking the moving sperm, an open loop control strategy can be used, and a closed loop control strategy can be used, so that the tip of the micropipette can rapidly, accurately and stably track the sperm in an image plane.
In an open loop control strategy, firstly determining the position of the tip of the micropipette in an image plane according to image feedback of the tip of the micropipette, then lifting the tip of the micropipette in the normal direction of the image plane by using a positioner, and adjusting the position of the tip of the micropipette in an open loop manner according to the real-time position of a mobile sperm so as to realize sperm tracking; in a closed-loop strategy, the positions of the tip of the micropipette and the sperm are observed in real time on a focal plane, closed-loop motion control is performed, and meanwhile passive motion of the sperm to be operated caused by disturbance of a liquid substrate by a moving sperm operating tool is compensated, so that the process of tracking the sperm to be operated by the tip of the sperm operating tool is quickened.
Motion compensation of the micropipette tip for liquid substrate disturbances is specifically implemented as:
wherein, the liquid crystal display device comprises a liquid crystal display device,
controlling the axial speed of the operating tool for the positioner, < >>
Axial disturbance model of the liquid substrate for the operation of the tool movement, +.>
For the estimation of unknown parameters in the axial disturbance model, epsilon is the axial unit vector,e=p s -p t =[x y] T for tracking error +.>
And->
Is of normal number>
The gain is determined positively. />
For the positioner to control the speed of the operating tool in the axial vertical direction, +.>
For operating a disturbance model of the tool movement in the axial vertical direction of the liquid substrate, +.>
For the estimation of unknown parameters in the disturbance model in the axial vertical direction +.>
As a unit vector in the axial vertical direction,
and->
Is of normal number>
The gain is determined positively.
S3, using a positioner to enable the tip of the sperm operating tool and the mobile sperm to remain relatively static in an image plane and move normally in the image plane, and realizing sperm braking through physical interaction between the tip of the sperm operating tool and the mobile sperm.
The motion speed and the motion direction of the sperm at the current moment are predicted by an extended Kalman filtering algorithm, and the motion of the tip of the micropipette is controlled by a positioner based on the prediction, so that the tip and the moving sperm are kept relatively static in an image plane.
At the same time, the micropipette tip is controlled by the positioner to move toward the liquid substrate base at 2000 microns per second normal to the image plane until it comes into contact with the liquid substrate base. At this time, the motor protein at the tail of the sperm is deactivated due to the pressing of the tip of the micropipette, so that the sperm cannot move, and the effect of sperm braking is achieved.
S4, adjusting the pressure in the cavity of the sperm operating tool by using a pressure device, and automatically sucking and positioning the braking sperm.
In a specific embodiment, the tip of the micropipette is placed at the braked sperm, and the pressure device is used to adjust the pressure in the micropipette cavity to negative pressure, thereby sucking the liquid substrate containing the braked sperm. When the sperm to be operated enters the micropipette, if the sperm does not reach the designated position, the pressure device is used for keeping the negative pressure in the micropipette, so that more liquid substrates are sucked, and the braked sperm moves towards the designated position. Conversely, when the braked sperm passes over the designated location of the micropipette, the pressure device will adjust the pressure within the chamber of the micropipette to a positive pressure to move and hold the braked sperm in the designated location within the micropipette while the excess liquid substrate is being expelled.
As shown in fig. 3, based on the same inventive concept, the present invention also provides a system for implementing the sperm automatic braking and sucking method as described above, the system comprising: a table 10, an image data display unit 20, an image data acquisition unit 30, a micropipette unit 40, a host unit 50, and a base 60. The table 10, the image data display unit 20, the image data acquisition unit 30, the micropipette unit 40, and the host unit 50 are provided on a base 60.
Wherein the stage 10 is provided for placing a liquid substrate containing sperm, the stage 10 is placed within the imaging range of the image data display unit 20.
The image data display unit 20 is used for imaging sperm, and the image data acquisition unit 30 and the image data display unit 20 perform image transmission through an optical path. In one embodiment, the image data display unit 20 is a microscope, and the focal length of the microscope is controllable by the host unit 50.
The image data acquisition unit 30 is used for acquiring the sperm image imaged by the image data display unit 20, and may send the acquired image data to the host unit 50 for processing. In one embodiment, the image data acquisition unit 30 is a CCD camera.
The micropipette unit 40 is configured to perform a sperm cell operation and includes a positioner 41, the positioner 41 being coupled to a sperm cell operation tool 42, the operation end of the sperm cell operation tool 42 extending above the platen 10. Sperm manipulation tool 42 is a micropipette having a hollow needle-shaped tip. The positioner 41 holding the micropipette is operably connected to the host unit 50 while the table 10 supporting the liquid substrate containing sperm need not be operably connected to the host unit 50.
In addition, in order to perform sperm manipulation, micropipette unit 40 further includes a connecting tubing 43 and a pressure pump 44; the pressure pump 44 is connected to the micropipette via a connecting conduit 43, so that the pressure pump 44 can regulate the pressure in the micropipette chamber. In this way, the pressure pump 44 is operated to allow the micropipette to aspirate sperm through the connecting tubing 43.
The host unit 50 transmits data to the image data acquiring unit 30, and processes the image data sent by the image data acquiring unit 30 to obtain the distance between the sperm and the micropipette in the image data, and the host unit 50 controls the positioner 41 to drive the sperm manipulation tool 42 to perform braking and sucking actions according to the distance.
In summary, the present invention provides a method for automatically braking and sucking sperm by manipulating a micropipette for the first time without using a controllable sperm liquid substrate table. Thanks to the sperm scoring mechanism and the motion control strategy of the invention, the invention can autonomously select or assist an operator to rapidly select sperm in the field of view suitable for operation; turbulence of the sperm cell process tool motion on the sperm cell containing liquid substrate can be avoided or compensated for. The invention simplifies the constitution of the sperm operation system, is more close to the current equipment conditions of most hospitals, and reduces the development cost of the sperm operation system.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential characteristics thereof. The present embodiments are, therefore, to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present disclosure describes embodiments, not every embodiment is provided with a separate embodiment, and that this description is provided for clarity only, and that the disclosure is not limited to the embodiments described in detail below, and that the embodiments described in the examples may be combined as appropriate to form other embodiments that will be apparent to those skilled in the art.
Claims (7)
1. An automatic sperm braking and sucking method, which is characterized by comprising the following steps:
s1, placing a liquid substrate containing sperms on a workbench, enabling the liquid substrate to be located in an imaging range of an image data display unit, acquiring images of the tip of a sperm operation tool, sperms and the liquid substrate by using an image data acquisition unit, and transmitting the images to a host unit;
the step S1 also comprises the step of automatically selecting sperms to be operated by a system or actively selecting sperms to be operated by an operator; the method further comprises the steps of generating each sperm fraction according to the expected movement track of each sperm, the orientation of each sperm and the distance between each sperm and the center of the visual field through visual feedback of the sperm and the liquid substrate so as to realize automatic or auxiliary active selection of the sperm to be operated by a system operator; the scoring method comprises the following steps:
c＝[c 1 …c 4 ]·[w 1 …w 4 ] T (1)
c 3 ＝-k 3 ln(|p s -p c |+ε 3 ) (5)
c 4 ＝-k 4 ln(|p s -p t |+ε 4 ) (6)
Wherein c is the comprehensive score of sperm to be processed, c 1 …c 4 W 1 …w 4 Independent scoring and weighting epsilon of four indexes of sperm to be operated i Where i=1, 3,4 is a positive constant, abs (·) represents absolute value, E (·) represents desired, z i For the ith predicted position of sperm to be manipulated, θ i Representing the likelihood of step i, p c ，p s ，p t The positions of sperm and tip, k, respectively, at the center of the field of view 1 ,k 21 ,k 22 ,k 3 ,k 4 Is gain;
s2, the host unit controls the sperm operation tool through the control locator according to the distance between the sperm operation tool tip and the sperm in the image, so that the sperm operation tool tip tracks and moves the sperm in the direction parallel to the image plane until the sperm operation tool tip and the sperm are positioned at the same position in the image plane;
the step S2 further comprises a step of tracking sperm in the image plane, comprising:
the open loop method comprises the following steps: firstly, determining the position of the sperm in an image plane according to image feedback of the tip of a sperm operation tool, then using a positioner to normally lift the tip of the sperm operation tool in the image plane, and controlling the position of the tip of the sperm operation tool according to a moving sperm real-time image in an open loop manner to realize sperm tracking; or:
the closed loop method comprises the following steps: observing the positions of the tip of the sperm manipulation tool and the sperm in real time on a focal plane, performing closed-loop motion control, and compensating for passive motion of the sperm to be manipulated caused by disturbance of a liquid substrate by the moving sperm manipulation tool so as to accelerate the tip of the sperm manipulation tool to track the sperm to be manipulated;
motion compensation of the micropipette tip for liquid substrate disturbances is specifically implemented as:
wherein v is x For the positioner to control the axial speed of the operating tool,
axial disturbance model of the liquid substrate for the operation of the tool movement, +.>
For estimation of unknown parameters in an axial disturbance model, ε is an axial unit vector, e=p s -p t ＝[xy] T For tracking error, k px And K is equal to ex Is a normal number Γ x To positively set the gain, v y For the positioner to control the speed of the operating tool in the axial vertical direction, +.>
For operating a disturbance model of the tool movement in the axial vertical direction of the liquid substrate, +.>
For estimation of unknown parameters in a disturbance model in an axially perpendicular direction, delta is a unit vector in the axially perpendicular direction, k py And k is equal to ey Is a normal number Γ y Gain is positively determined;
s3, using a positioner to enable the tip of the sperm operating tool and the mobile sperm to remain relatively static in an image plane and move normally in the image plane, and realizing sperm braking through physical interaction between the tip of the sperm operating tool and the mobile sperm;
s4, adjusting the pressure in the cavity of the sperm operation tool, and automatically sucking and positioning the braking sperm.
2. The method according to claim 1, wherein the method further comprises:
updating the fraction of each sperm in real time according to the changes of the direction, the distance and the expected movement track caused by the movement of each sperm, and considering the switching cost of an operation target generated by the replacement of the sperm in the operation process, so as to replace the sperm to be operated when necessary;
the method further comprises the steps of:
the switching cost of the operation target is considered in the process of automatically selecting and replacing the sperm to be operated so as to avoid frequent replacement of the sperm to be operated.
3. The method according to claim 1, wherein the step S3 further comprises:
predicting and compensating the movement of the mobile sperm to reduce the relative error of tracking the mobile sperm, and stopping the sperm to be operated by moving the sperm cell manipulator tip normal to the image plane to contact the bottom of the liquid substrate and squeeze the mobile sperm.
4. A system for implementing the sperm automatic braking and aspiration method of any one of claims 1-3, said system comprising: the device comprises a workbench, an image data display unit, an image data acquisition unit, a micropipette unit and a host unit;
the workbench is arranged in the imaging range of the image data display unit;
the image data acquisition unit and the image data display unit perform image transmission through an optical path;
the micropipette unit comprises a locator connected to a sperm manipulation tool;
the operation end of the sperm cell operation tool extends to the upper part of the workbench;
the host unit is in data transmission with the image data acquisition unit, and processes the image data sent by the image data acquisition unit to obtain the distance between sperms in the image data and the operating end of the sperm operating tool, and the host unit controls the locator to drive the sperm operating tool to brake and absorb according to the distance.
5. The system of claim 4, wherein the sperm cell manipulation tool is a micropipette having a hollow needle-like tip.
6. The system of claim 4 or 5, wherein the micropipette unit further comprises a pressure mechanism in communication with the sperm cell manipulator means through a conduit, and wherein the pressure mechanism is controlled by the host unit to cause the sperm cell manipulator means to perform a pipetting action.
7. The system of claim 6, wherein the pressure mechanism is a pressure pump.
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| WO2012037642A1 (en) * | 2010-08-20 | 2012-03-29 | Marksmen Cellject Inc. | System and method for automated sperm manipulation |
| WO2015153691A2 (en) * | 2014-04-03 | 2015-10-08 | Drexel University | Computer-implemented methods, computer-readable media, and systems for tracking a plurality of spermatozoa |
| GB201808312D0 (en) * | 2018-05-21 | 2018-07-11 | Governing Council Of The Univ Of Toronto | A method for automated non-invasive measurement of sperm motility and morphology and automated selection of a sperm with high dna integrity |
| WO2021014461A1 (en) * | 2019-07-19 | 2021-01-28 | Payeli Sravan Kumar | Intracytoplasmic sperm injection |
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