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WO2021014461A1 - Injection intra-cytoplasmique de sperme - Google Patents

Injection intra-cytoplasmique de sperme Download PDF

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Publication number
WO2021014461A1
WO2021014461A1 PCT/IN2020/050610 IN2020050610W WO2021014461A1 WO 2021014461 A1 WO2021014461 A1 WO 2021014461A1 IN 2020050610 W IN2020050610 W IN 2020050610W WO 2021014461 A1 WO2021014461 A1 WO 2021014461A1
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WIPO (PCT)
Prior art keywords
pressure value
oocyte
syringe
pressure
micropipette
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PCT/IN2020/050610
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English (en)
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Sravan Kumar PAYELI
Pavani Srividya MOCHARLA
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M21/00—Bioreactors or fermenters specially adapted for specific uses
    • C12M21/06—Bioreactors or fermenters specially adapted for specific uses for in vitro fertilization
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M35/00—Means for application of stress for stimulating the growth of microorganisms or the generation of fermentation or metabolic products; Means for electroporation or cell fusion
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12M—APPARATUS FOR ENZYMOLOGY OR MICROBIOLOGY; APPARATUS FOR CULTURING MICROORGANISMS FOR PRODUCING BIOMASS, FOR GROWING CELLS OR FOR OBTAINING FERMENTATION OR METABOLIC PRODUCTS, i.e. BIOREACTORS OR FERMENTERS
    • C12M41/00—Means for regulation, monitoring, measurement or control, e.g. flow regulation
    • C12M41/48—Automatic or computerized control

Definitions

  • the present subject matter relates to intracytoplasmic sperm injection, and particularly relates to automation and real-time monitoring of intracytoplasmic sperm injection, pattern recognition corresponding to pressure applied on oocytes during the intracytoplasmic sperm injection, and prediction of oocyte quality after the intracytoplasmic sperm injection.
  • IVF Invitro fertilization
  • ICSI Intracytoplasmic sperm injection
  • a micromanipulator In ICSI procedure, a micromanipulator is provided with an oocyte holding handle and a sperm injection handle, that are connected to syringes for providing aspiration pressure for holding oocyte and puncturing oocyte with an ejection pressure to deposit the sperm, respectively.
  • joysticks are used in the micromanipulator for micro controlling of the holding and injection needles simultaneously for oocyte manipulation.
  • FIG. 1 A illustrates a system for performing real-time monitoring of ICSI, in accordance with an implementation of the present subject matter
  • FIG. 1 B illustrates an expanded vertical sectional view of the droplets in the dish as shown in Fig. 1 A, with an ovum handling medium microdroplet and a microdroplet of polyvinylpyrrolidone (PVP) for sperm in the dish, in accordance with an implementation of the present subject matter;
  • PVP polyvinylpyrrolidone
  • FIG. 2 illustrates a system for real-time monitoring and automation of ICSI, in accordance with an implementation of the present subject matter
  • FIG. 3A to 3H illustrate an arrangement of a holding needle and an injection needle for performing ICSI process, in accordance with an implementation of the present subject matter
  • FIG. 4 illustrates graphical representation of the readings of holding pressure and injection pressure for real-time ICSI monitoring, in accordance with an implementation of the present subject matter
  • FIG. 5 illustrates graphical representation of biomechanical property measurement and pattern recognition, in accordance with an implementation of the present subject matter.
  • a system for performing the ICSI process requires that an operator, referred to as an embryologist, apply manual aspiration pressure on holding needle to hold the oocyte and a mechanical pressure on an injection needle.
  • the application of manual pressure may be to press the injection needle on an oocyte which is placed in an oocyte handling liquid medium.
  • the manual application of push pressure allows the injection needle to penetrate the first layer (zona pellucida) of the oocyte.
  • an aspiration pressure is applied by manually actuating the injection syringe to puncture a second layer (oolemma) of the oocyte.
  • a positive pressure is applied from the injection needle by actuating the injection syringe to deposit a sperm from the injection needle into the cytoplasm.
  • the present subject matter describes systems for automation and real-time monitoring of the ICSI process.
  • the present subject matter also describes pattern recognition corresponding to pressure applied on oocytes during the ICSI process, and prediction of oocyte quality after the ICSI process on day 0 (the day on which the ICSI procedure is performed).
  • force or pressure sensors may be connected with a holding needle and an injection needle. These pressure sensors may measure the values of the pressure applied on the injection needle side as well as on the holding needle side.
  • the system described herein utilizes the measured values and implements data visualization techniques for real-time monitoring of the ICSI process.
  • the measured values from the respective pressure sensor from the holding needle side and the injection needle side are displayed on an ICSI monitor as a histogram, for the purpose of real time monitoring of a number of sperms collected, the pressure value applied to hold the oocyte, a number of oocytes processed with sperm injection, time taken for the sperm injection process, and a value pertaining to the elasticity of the first layer of the oocyte.
  • the system described herein may be provided with a machine learning framework including a neural network.
  • the neural network may be involved in the training of a model of the machine learning framework.
  • the pressure values measured from the pressure sensors, along with individual outcomes of blastocyst formation as a result of respective sperm injection may be provided to the neural network.
  • the trained model may then provide an output with the pressure values required for at the holding needle side and the injection needle side in order to automate the system.
  • the automation may involve implementing the output pressure values to be applied on the injection needle side and the holding needle side.
  • Automation of various steps performed by the system, as per the present subject matter, may require relatively less time to perform the ICSI process as compared to the manually operated conventional systems.
  • FIG. 1A illustrates a system 100 for real-time monitoring of ICSI in accordance with an implementation of the present subject matter.
  • the system 100 includes a micromanipulator 102 and a microscopic lens (not shown).
  • the micromanipulator 102 may include a heating surface 104 on which a dish 106 may be placed.
  • Oocytes may be placed in a culture medium microdroplet 108 in the dish 106.
  • Sperms may be placed in a polyvinylpyrrolidone (PVP) microdroplet 1 10 in the dish 106, separate from the culture medium microdroplet 108, for immobilization of the sperms.
  • the heating surface 104 may maintain respective temperatures of the culture medium microdroplet 108 and the PVP microdroplet 1 10 in the dish 106 at a predetermined temperature, during the ICSI process.
  • the predetermined temperature may be 37 degrees Celsius.
  • a holding needle 112 may be placed in a first holder 1 14.
  • the holding needle may also be referred hereinafter as a first micropipette.
  • the first holder 1 14 may be connected to a first syringe 1 16 with a first flexible tube 1 18.
  • the first syringe 1 16 may control positive and negative pressures applied in the holding needle 1 12 based on the movement of a piston of the first syringe 1 16.
  • a first pressure sensor 120 may be connected between the first syringe 1 16 and the first holder 1 14 through a first fixture 122.
  • the first holder 1 14 may be placed over the dish 106 with the help of a first rig 124 of the micro-manipulator.
  • the holding needle 112 may be horizontally aligned in the dish 106 and is used to hold an oocyte to be injected with a sperm.
  • An injection needle 126 may be placed in a second holder 128.
  • the injection needle 126 may also be referred hereinafter as a second micropipette.
  • the second holder 128 may be connected with a second syringe 130 with a second flexible tube 132.
  • the second syringe 130 may control positive and negative pressures applied in the injection needle 126 based on the movement of a piston of the second syringe 130.
  • a second pressure sensor 134 may be connected between the second syringe 130 and the second holder 128 through a second fixture 136.
  • the second holder 128 may be placed over the dish 106 with the help of a second rig 138 of the micromanipulator 102.
  • the sperms to be injected in the oocytes are collected into the injection needle 126, through a controlled aspiration pressure applied in the injection needle with the second syringe 130.
  • the first pressure sensor 120 and the second pressure sensor 134 may be connected to an ICSI monitor 142.
  • the ICSI monitor 142 may also be referred hereinafter as a display unit.
  • the holding needle 1 12 and the injection needle 126 may be aligned along an axis 140, and in close proximity to each other (as shown in FIG. 3A).
  • the injection needle 126 may be aligned such that an opening end of the injection needle 126 faces an opening end of the holding needle 112.
  • the injection needle 126 may be pushed towards the holding needle 1 12, into 30% to 70% of the oocyte diameter, using a mechanical displacement pressure, due to the elastic behavior of oocyte (as shown in FIG. 3B).
  • the mechanical push pressure may be applied until the injection needle 126 may penetrate through an outer layer (zona pellucida) of the oocyte (as shown in FIG. 3C and 3D).
  • an aspiration pressure may be applied on the injection needle 126 through the second syringe 130 in order to puncture an inner layer (oolemma) of the oocyte (as shown in FIG. 3E and 3F).
  • the puncturing of the oolemma is visually observed or evidenced by a sharp inflow of ooplasm of the oocyte into the injection needle 126, through the microscopic lens. Viscosity of the cytoplasm of the oocyte is significantly lower than the viscosity of the oolemma, which results in the sharp inflow of the ooplasm after the successful puncturing of the oolemma and a pressure drop.
  • a quick positive pressure is applied on the injection needle 126 through the second syringe 130 to neutralize the applied aspirated pressure (as shown in FIG. 3G).
  • an additional positive pressure is applied on the injection needle 126 to deposit the sperm into the ooplasm (as shown in FIG. 3H).
  • the injection needle 126 is taken out from the oocyte, and the oocyte, deposited with the sperm, is released from the holding needle 1 12 by releasing the applied pressure on the first syringe 1 16.
  • a pattern of pressure values measured at the holding needle side and the injection needle side, measured by the first pressure sensor 120 and the second pressure sensor 134, may be computed. Further, the pattern may be used to plot a histogram which is displayed on the ICSI monitor 142. The displaying of the histogram on the ICSI monitor 142 is to allow real-time monitoring of number of oocytes injected with sperms, time taken for sperm injection per oocyte, and pressure applied to hold the oocyte.
  • a machine learning framework may be utilized for performing a prediction of oocyte quality in which training data which may be provided to a computing system (not shown) for performing the prediction.
  • the training data may be provided in the form of a blastocyst formation data which is recorded in a blastocyst formation phase with image processing, and the respective oocyte elasticity pattern data which are fed into the computing system by the user.
  • the machine learning framework may train a model, based on the user fed data, for prediction of the quality of oocytes immediately after the ICSI process based on biomechanical properties as measured in the current subject matter.
  • the pressure patterns and the blastocyst observation data, collected and provided at the machine learning framework, are reflective of the oolemma elasticity, the internal health and quality of the oocyte.
  • This data may be fed into the computing system including a software, and a code is generated.
  • the generated code may be used to allow the operation of the second syringe 130 for sperm deposition upon successful puncture of oolemma by applying a required pressure.
  • the oocyte is held by the holding needle 112 with an aspiration pressure (negative pressure) in a range from 1 kilo-pascal (kPa) to 3 kPa.
  • the atmospheric pressure is in a range of 90 kPa to 100 kPa depending on sea level and altitude.
  • the real-time monitoring of the holding pressures may allow for determining if higher pressure than required is applied on the holding needle side. Application of higher pressure than required for holding the oocyte may cause unwanted stress on oocyte. This unwanted stress on oocyte can be avoided by generation of an alarm signal by a software to alert the user, as the user at a particularly period of time would be focused on sperm deposition process.
  • the real-time monitoring of the oocyte holding pressure may allow for avoiding inter-user dependent variations in applying holding pressure for holding the oocyte.
  • the oocytes may undergo the process of fertilization, forming fertilized eggs (zygote).
  • the eggs are segregated and cultured for five days in an incubator for embryo development.
  • the fertilized eggs undergo a blastocyst formation phase.
  • the blastocyst formation of the eggs is observed and recorded through embryo grading or image processing on the fifth day of culturing of the eggs.
  • the recorded blastocyst formation data are correlated with the respective oocyte elasticity patterns to determine the quality of the oocyte.
  • FIG. 1 B illustrates expanded side sectional view of a dish 106 containing a culture medium microdroplet 108 containing oocytes which are to be inserted by sperms, and a PVP microdroplet 1 10 used for the immobilization of the sperms.
  • FIG. 2 illustrates a system 200 for real-time monitoring of ICSI process and automation of ICSI process, in accordance with an implementation of the present subject matter.
  • the system 200 may include a micromanipulator 202, a microscopic lens (not shown), a first syringe 204, a second syringe 206, a processing unit 208, an actuator 210, a first rig 212, a second rig 214, an imaging unit 216, and an image processor 218.
  • the micromanipulator 202 may include a heating surface 220. Further, a dish 222 may be placed on the heating surface 220. The dish 222 may contain a culture medium microdroplet 224 and a PVP microdroplet 226.
  • the functions of the heating surface 220, the dish 222, the culture medium microdroplet 224 and the PVP microdroplet 226 is similar to the functions of the heating surface 104, the dish 106, the culture medium microdroplet 108, and the PVP microdroplet 110 of FIG. 1A.
  • the first syringe 204 may be connected to a first micropipette 228.
  • the second syringe 206 may be connected to a second micropipette
  • the processing unit 208 may be communicatively coupled the actuator 210, the first syringe 204, the second syringe 206, and image processor 218.
  • the processing unit 208 may store a first pressure value, a second pressure value, a third pressure value, and a fourth pressure value.
  • the first pressure value, the second pressure value, the third pressure value, and the fourth pressure value may be based on a machine learning framework.
  • the first pressure value may be obtained from a database (not shown) and pertains to a negative pressure required for collecting the sperm into the second micropipette 230.
  • the database may store the first pressure value obtained by training a model of the machine learning framework over a plurality of values pertaining to the negative pressure applied for sperm collection.
  • the model may be trained based on a neural network of the machine learning framework by inputting pressure values pertaining to the pressure required collecting sperms from the dish 222.
  • the second pressure value may be obtained from the database based on a minimum pressure required to hold the oocyte.
  • the database may store the second pressure value obtained by training a model of the machine learning framework over a plurality of values.
  • the plurality of values may pertain to the pressure which is required to hold the oocyte at a tip of the first micropipette 228.
  • the third pressure value may be obtained from the database based on an elasticity of the second layer of the oocyte.
  • the database may store the third pressure value obtained by training a model of the machine learning framework over a plurality of values pertaining to the elasticity of the second layer of the oocyte.
  • the fourth pressure value may be obtained from the database based on a viscosity of cytoplasm of the oocyte.
  • the database may store the fourth pressure value obtained by training a model of the machine learning framework over a plurality of values which pertain to a confirmation of the puncturing of the second layer.
  • the plurality of values may pertain to the viscosity of the cytoplasm of the oocyte.
  • each of the first pressure value, the second pressure value, and the third pressure value is a negative pressure value
  • the fourth pressure value is a positive pressure value
  • the system 200 may include a first holder 232 and a second holder 234.
  • the first holder 232 may be provided for holding the first micropipette 228.
  • the first holder 232 may be coupled with the first syringe 204.
  • the second holder 234 may be provided for holding the second micropipette 230.
  • the second holder 234 may be coupled with the second syringe 206.
  • the first holder 232 may be coupled with the first syringe 204 via a first flexible tube 236.
  • the first syringe 204 may control positive and negative pressures provided to the first micropipette 228.
  • the second holder 234 may be coupled with the second syringe 206 via a second flexible tube 238.
  • the second syringe 206 may control positive and negative pressures provided to the second micropipette 230 for injecting an oocyte, puncturing oolemma and depositing the sperm, respectively.
  • the system 200 may further include a first pressure sensor
  • the first pressure sensor 240 may be connected between the first syringe 204 and the first holder 232 through a first fixture 244.
  • the first pressure sensor 240 may be connected between the first micropipette 228 and the first syringe 204.
  • the first pressure sensor 240 may be arranged to measure, in real-time, the second pressure value.
  • the second pressure sensor 242 may be connected between the second syringe 206 and the second holder 234 through a second fixture 246.
  • the second pressure sensor 242 may be connected between the second micropipette 230 and the second syringe 206.
  • the second pressure sensor 242 may be provided to measure the first pressure value, the third pressure value, and the fourth pressure value.
  • the first rig 212 may be arranged to hold the first micropipette 228 in place and to perform alignment of the first micropipette 228 as per requirement.
  • the second rig 214 may be arranged to hold the second micropipette 230 in place and to perform alignment of the second micropipette 230 as per requirement.
  • the first rig 212 and the second rig 214 may communicate with the actuator 210 which may control a respective position of the first rig 212 and the second rig 214.
  • the imaging unit 216 may be arranged in order to capture a first image of a sperm collection region.
  • the sperm collection region may be a section of the dish 222 at which the PVP microdroplet 226 may be present.
  • the image processor 218 may process the first image to determine a plane of focus pertaining to a target body in the image.
  • the target body includes a to-be collected sperm.
  • the image processor 218 may transmit a signal including the information pertaining to the determined plane of focus of the to-be injected sperm, to the processing unit 208.
  • the processing unit 208 may then control the actuator 210, based on the transmitted signal.
  • the actuator 210 by the control of the processing unit 208, may then actuate the second rig 214 to align a plane of focus pertaining to a tip of the second micropipette 230 to the plane of focus pertaining to the to-be injected sperm. The actuator 210 may then position the tip in proximity to the to-be injected sperm.
  • the actuator 210 may operate the second syringe 206 based on the first pressure value to collect a sperm from the culture medium microdroplet 224 into the second micropipette 230.
  • the imaging unit 216 may also be arranged in order to capture a second image of a sperm injection region.
  • the sperm injection region may be a section on the dish 222 at which the culture medium, containing oocytes, is present.
  • the image processor 218 may process the second image to determine a plane of focus pertaining to a target body in the image.
  • the target body includes a target oocyte.
  • the image processor 218 may transmit a signal including the information pertaining to the determined plane of focus of the target oocyte, to the processing unit 208.
  • the processing unit 208 may then control the actuator 210, based on the transmitted signal.
  • the actuator 210 by the control of the processing unit 208, may then actuate the first rig 212 to align a plane of focus pertaining to a tip of the first micropipette 228 to the plane of focus pertaining to the target oocyte.
  • the actuator 210 may then position the tip in proximity to the target oocyte.
  • the imaging unit 216 may be a camera unit. In an example, the imaging unit 216 may be placed over the dish 222 and an optical sensor (not shown) of the imaging unit 216 may be positioned to face towards the dish 222.
  • the imaging unit 216 may capture the first image and the image processor 218 may process the first image to determine a horizontal plane at which the to-be injected sperm is present. Similarly, the imaging unit 216 may process the second image to determine a horizontal plane at which the target oocyte is present.
  • the imaging unit 216 may be placed on a side of the dish 222 and an optical sensor of the imaging unit 216 may be positioned to face towards the dish 222.
  • the imaging unit 216 may capture the first image and the image processor 218 may process the first image to determine a vertical plane at which the to-be injected sperm is present.
  • the imaging unit 216 may process the second image to determine a vertical plane at which the target oocyte is present.
  • the dish 222 may be movably placed on a platform of the micromanipulator 202 and the imaging unit 216 may be stationary to the micromanipulator 202.
  • the dish 222 may be moved by the help of an actuating unit (not shown) to order to allow the imaging unit 216 to capture the first image and the second image.
  • the imaging unit 216 may be movably attached on the micromanipulator 202.
  • the dish 222 may be stationary to the micromanipulator 202, and the imaging unit 216 may be moved by the help of an actuating unit (not shown) to order to capture the first image and the second image.
  • the actuator 210 may operate the first syringe 204 based on the second pressure value to hold the oocyte at a tip of the first micropipette 228. Based on the second image, the actuator 210 may align the first micro pipette and the second micropipette 230 at an axis 248. The actuator 210 may then actuate the second micropipette 230 to penetrate a first layer of the oocyte.
  • the first layer may be an outer layer of the oocyte.
  • the outer layer of the oocyte is zona pellucida.
  • the actuator 210 may actuate a linear movement of the second micropipette 230 towards the oocyte for the second micropipette 230 into 30% to 70% of the oocyte diameter, due to the elastic behavior of oocyte, until the first layer is penetrated. After successful penetration of the first layer of the oocyte, the actuator 210 may operate the second syringe 206 based on the third pressure value to puncture a second layer of the oocyte via the second micropipette 230.
  • the second layer may be an inner layer of the oocyte.
  • the inner layer is oolemma. The elasticity of the inner layer in higher than the elasticity of the outer layer.
  • the puncturing of the oolemma may be visually observed or evidenced by a sharp inflow or a pressure drop of ooplasm of the oocyte into the first micropipette 228, through a microscope.
  • the actuator 210 may operate the second syringe 206 in order to apply a predetermined positive pressure in order to neutralize the aspiration pressure applied to puncture the second layer, as soon as the puncturing of the oolemma occurs.
  • the actuator 210 may operate the second syringe 206 based on the fourth pressure value to deposit the collected sperm into the oocyte. After the successful deposition of the sperm into the oocyte, the first micropipette 228 may be taken out from the oocyte by the actuator 210, and the oocyte, deposited with the sperm, may be released from the second micropipette 230 by application of a positive pressure through the first syringe 204.
  • the system 200 may further include a display unit 250 which may be coupled with the first pressure sensor 240 and the second pressure sensor 242.
  • the first pressure sensor 240 and the second pressure sensor 242 may provide the measured first, second, third, and fourth pressure values to the display unit 250.
  • the display unit 250 may be provided to display the measured first, second, third, and fourth pressure values, simultaneously, on a single plot in real-time.
  • the function of the display unit 250 may be similar to the ICSI monitor 142 of FIG. 1 A.
  • the display unit 250 may display the measured pressure values as a histogram for real-time monitoring of a number of sperms collected, the pressure value applied to hold the oocyte, a number of oocytes processed with sperm injection, time taken for the sperm injection process, and a value pertaining to the elasticity of the first layer of the oocyte.
  • the first pressure sensor 240 and the second pressure sensor 242 may be coupled with the display unit 250 to provide pressure values to the ICSI monitor 142 for displaying the pressure values on the screen of the ICSI monitor 142.
  • FIG. 4 illustrates graphical representation of the readings of holding pressure and injection pressure for real-time ICSI monitoring, in accordance with an implementation of the present subject matter.
  • the readings of holding pressure and injection pressure are acquired as per the description provided for FIG. 1 A.
  • a horizontal axis of the graph 400 shows time (in seconds) taken for performing the ICSI process.
  • the vertical axis of the graph 400 shows the pressure (in kPA) being applied at the holding needle side and the injection needle side.
  • first values 402 pertains to the pressure being applied at the holding needle side and second values 404 pertains to the pressure being applied at the injection needle side.
  • Blocks 406, 408, and 410 represents the duration for individual sperm injections been performed.
  • the pressure applied in the holding needle side and the injection needle side may be measured with the first pressure sensor and the second pressure sensor, respectively.
  • the first pressure sensor and the second pressure sensor may be connected to the ICSI monitor where the measured pressure values from the first pressure sensor and the second pressure sensor are displayed on the ICSI monitor as a histogram through an ICSI visualization software.
  • at least 5 data points per second, representing the pressure values measured at the holding needle side are recorded.
  • 20 to 50 data points per second, representing the pressure values measured at the injection needle side, per second are recorded.
  • the data points, recorded at the holding needle side and the injection needle side, may be plotted with respect to time in real-time, and the plot may be displayed in the ICSI monitor in order to understand the oolemma viscoelastic properties.
  • the integration of the data into the ICSI visualization software for real-time monitoring allows the user to easily determine the number of oocytes injected with sperms, and the pressures applied at holding and injection needle sides with respect to time.
  • FIG. 5 illustrates graphical representation of biomechanical property measurement and pattern recognition, in accordance with an implementation of the present subject matter.
  • the readings of injection pressure as represented in FIG. 5, are acquired as per the description provided for FIG. 1A.
  • the readings are associated with the sperm injection process, represented in the graph 500 as Run 1 , Run 2, and Run 3, performed on three different oocytes injected with three individual sperms.
  • a horizontal axis of the graph 500 shows time (is seconds) taken for performing the ICSI process.
  • the vertical axis of the graph 500 shows the pressure (in kPA) dynamics at the injection needle side during oolemma puncturing and sperm deposition events.
  • Block 502 represents the values defining oolemma elasticity pertaining to each of the oocytes related to the Run 1 , Run 2, and Run 3, respectively. Referring to block 502, the value of oolemma elasticity is 1 1.3 kPA/ 0.8 sec.
  • Block 504 represents the values defining oolemma plasticity pertaining to the three oocytes. Referring to block 504, the value of oolemma plasticity is 10 kPA/ 0.7 sec.
  • Block 506 represents the values defining cytosol viscosity pertaining to the three oocytes or dynamics of pressure drop that corresponds to a cytoplasmic content nature or viscous behavior. Referring to block 506, the value of cytosol viscosity is 10 kPA/ 0.6-0.7 sec.
  • the sharp inflow of the ooplasm or the neutralization of the negative pressure after successful puncturing of the oolemma may be recorded from the real-time readings from the ICSI monitor.
  • a pattern of the pressure values measured at the injection needle side represent the oolemma elasticity and ooplasm viscosity.
  • a pattern of pressure values measured at the holding needle side may be used for real time monitoring of number of oocytes injected with sperms, time taken for sperm injection per oocyte, and pressure applied to hold the oocyte.
  • a code may be generated, by a computing unit, coupled with the pressure sensors, from the recorded values of ooplasm inflow or negative pressure neutralization to read the injection needle side pressure patterns.
  • the injection needle side pressure patterns may be used to determine the oocyte elasticity patterns. Biomechanical properties (viscoelastic property) of each oocyte, injected with a sperm, may be measured and recorded.
  • the approaches of the present subject matter provide real time monitoring of number of oocytes held and injected, which avoids human errors, real-time pattern recognition of oocyte biomechanical properties during sperm deposition, which allows the measurement of oocyte quality.
  • ICSI automation allows precise regulation of the process for oolemma puncture and sperm deposition by applying either aspiration pressure or positive pressure based of the pressure patterns. ICSI automation allows for effective oolemma puncture and sperm deposition, while minimizing human error involved in the process of ICSI.

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Abstract

L'invention concerne un système (100, 200) d'injection intra-cytoplasmique de sperme automatisé Système (100, 200) comprenant des première et seconde seringues (116, 130, 204, 206) reliées à des première et seconde micropipettes 5 (112, 126, 228, 230), respectivement, une unité de traitement (208) pour stocker des première, deuxième, troisième et quatrième valeurs de pression, et un actionneur (210) couplé à l'unité de traitement (208), une première seringue (116, 204), et une seconde seringue (130, 206), pour faire fonctionner la seconde seringue (130, 206) première valeur de pression sur la base de la première valeur de pression pour collecter le sperme, une première seringue (116, 204) sur la base d'une seconde valeur de pression pour maintenir un ovocyte, une seconde micropipette (126, 230) pour pénétrer dans la première couche d'ovocyte, faire fonctionner une seconde seringue (130, 206) sur la base d'une troisième valeur de pression pour percer une seconde couche d'ovocyte, et actionner une seconde seringue (130, 206) sur la base d'une quatrième valeur de pression pour déposer du sperme dans l'ovocyte. Les valeurs de pression sont basées sur une structure d'apprentissage machine.
PCT/IN2020/050610 2019-07-19 2020-07-16 Injection intra-cytoplasmique de sperme Ceased WO2021014461A1 (fr)

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Cited By (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPWO2023063099A1 (fr) * 2021-10-14 2023-04-20
CN115992183A (zh) * 2023-03-21 2023-04-21 苏州博致医疗科技有限公司 精子自动制动与吸取方法及系统
CN118879485A (zh) * 2024-09-27 2024-11-01 季华实验室 Icsi辅助操作仪及其控制方法
WO2025006004A1 (fr) * 2023-06-26 2025-01-02 Conceivable Life Sciences Inc. Fécondation in vitro automatisée intelligente et arrière-plan de plateforme d'injection de sperme intracytoplasmique
US12245793B2 (en) 2023-06-26 2025-03-11 Conceivable Life Sciences Inc. Robotic microtool control in an intelligent automated in vitro fertilization and intracytoplasmic sperm injection platform

Non-Patent Citations (3)

* Cited by examiner, † Cited by third party
Title
K. K. TAN ET AL: "Robust computer-controlled system for intracytoplasmic sperm injection and subsequent cell electro-activation", INTERNATIONAL JOURNAL OF MEDICAL ROBOTICS AND COMPUTER ASSISTEDSURGERY, vol. 5, no. 1, 1 March 2009 (2009-03-01), GB, pages 85 - 98, XP055704198, ISSN: 1478-5951, DOI: 10.1002/rcs.239 *
YULIANG ZHAO ET AL: "A Review of Automated Microinjection of Zebrafish Embryos", MICROMACHINES, vol. 10, no. 1, 24 December 2018 (2018-12-24), pages 7, XP055705950, DOI: 10.3390/mi10010007 *
ZHE LU ET AL: "Robotic ICSI (Intracytoplasmic Sperm Injection)", IEEE TRANSACTIONS ON BIOMEDICAL ENGINEERING, IEEE SERVICE CENTER, PISCATAWAY, NJ, USA, vol. 58, no. 7, 1 July 2011 (2011-07-01), pages 2102 - 2108, XP011408440, ISSN: 0018-9294, DOI: 10.1109/TBME.2011.2146781 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPWO2023063099A1 (fr) * 2021-10-14 2023-04-20
EP4417700A4 (fr) * 2021-10-14 2025-04-23 Shimadzu Corporation Procédé d'évaluation d'ovule, dispositif d'évaluation d'ovule et programme d'évaluation d'ovule
CN115992183A (zh) * 2023-03-21 2023-04-21 苏州博致医疗科技有限公司 精子自动制动与吸取方法及系统
WO2025006004A1 (fr) * 2023-06-26 2025-01-02 Conceivable Life Sciences Inc. Fécondation in vitro automatisée intelligente et arrière-plan de plateforme d'injection de sperme intracytoplasmique
US12245793B2 (en) 2023-06-26 2025-03-11 Conceivable Life Sciences Inc. Robotic microtool control in an intelligent automated in vitro fertilization and intracytoplasmic sperm injection platform
US12478405B2 (en) * 2023-06-26 2025-11-25 Conceivable Life Sciences Inc. Centrifuge-free sperm preparation in an intelligent automated in vitro fertilization and intracytoplasmic sperm injection platform
CN118879485A (zh) * 2024-09-27 2024-11-01 季华实验室 Icsi辅助操作仪及其控制方法

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