CN110719952B - Flexible guiding piezoelectric drill device with large axial vibration and small transverse vibration - Google Patents
Flexible guiding piezoelectric drill device with large axial vibration and small transverse vibration Download PDFInfo
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- CN110719952B CN110719952B CN201880031544.0A CN201880031544A CN110719952B CN 110719952 B CN110719952 B CN 110719952B CN 201880031544 A CN201880031544 A CN 201880031544A CN 110719952 B CN110719952 B CN 110719952B
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- flexible member
- micropipette
- flexible
- oocyte
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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
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L3/00—Containers or dishes for laboratory use, e.g. laboratory glassware; Droppers
- B01L3/02—Burettes; Pipettes
- B01L3/021—Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids
- B01L3/0217—Pipettes, i.e. with only one conduit for withdrawing and redistributing liquids of the plunger pump type
- B01L3/022—Capillary pipettes, i.e. having very small bore
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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
- C12M1/00—Apparatus for enzymology or microbiology
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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
- C12M33/00—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus
- C12M33/04—Means for introduction, transport, positioning, extraction, harvesting, peeling or sampling of biological material in or from the apparatus by injection or suction, e.g. using pipettes, syringes, needles
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- B—PERFORMING OPERATIONS; TRANSPORTING
- B01—PHYSICAL OR CHEMICAL PROCESSES OR APPARATUS IN GENERAL
- B01L—CHEMICAL OR PHYSICAL LABORATORY APPARATUS FOR GENERAL USE
- B01L2400/00—Moving or stopping fluids
- B01L2400/04—Moving fluids with specific forces or mechanical means
- B01L2400/0403—Moving fluids with specific forces or mechanical means specific forces
- B01L2400/0433—Moving fluids with specific forces or mechanical means specific forces vibrational forces
- B01L2400/0439—Moving fluids with specific forces or mechanical means specific forces vibrational forces ultrasonic vibrations, vibrating piezo elements
Abstract
A flexible guided piezoelectric drill device that produces large axial vibrations and small lateral vibrations is described. The flexible mechanism serves to guide the trajectory of the micropipette in the axial direction and to remove the resonance frequencies causing large lateral vibrations from the drive pulse. The invention is particularly suitable for efficiently penetrating the Zona Pellucida (ZP) of oocytes and embryos with little distortion.
Description
Technical Field
The present invention relates to an inertial shock injection device and more particularly to applications in oocyte and embryo manipulation for in vitro fertilization.
Background
Penetrating the Zona Pellucida (ZP) is an important step In Vitro Fertilization (IVF), using for example intracytoplasmic single sperm injection (ICSI) and biopsy for pre-implantation genetic screening (PGS). ZP is a thick highly elastic layer consisting of glycoproteins around oocytes or embryos.
In IVF clinical oocyte and embryo handling, ZP penetration requires severe deformation of the oocyte or embryo (e.g., 50 μm) even with a sharp micropipette. Large oocyte deformations may damage the oocyte spindles and lead to failure of oocyte fertilization and embryo development. Large embryo deformations can also affect subsequent embryo development. Using piezoelectrically actuated vibrations may improve ZP penetration without causing large oocyte or embryo deformation. The power drill is an inertial percussion device that uses a piezoelectric actuator to generate high frequency axial vibration of the micropipette to cause local rupture of the ZP.
Current push-drill designs cause cell damage due to unwanted large lateral vibrations on the micropipette tip. To help suppress lateral vibrations, a section of mercury is typically used near the tip of the micropipette. The use of mercury in direct contact with biomaterials has attracted clinical practice and biological research concerns, which are major obstacles to the entry of existing piezoelectric drill devices into IVF clinics.
Commercial power drills from Prime Tech and Burleigh, et al are designed with the piezoelectric actuator located away from the micropipette and producing large lateral vibrations at the micropipette tip. Placing the piezoelectric actuator directly behind the micropipette holder may help to better focus the vibrations on the micropipette tip. It is reported that this design configuration reduces lateral vibration to about 20 μm (h.b. huang, h.su, h.chen, and j.k.mills, "piezo drive non-Piezoelectric injector for automated cell management," medical instruments visual responsiveness reference, page 231-. However, this design still causes a considerable lateral vibration of 20 μm and results in a deformation of the oocyte of more than 10 μm.
Prior patents relevant to the present invention include US6251658B1 by Burleigh Instruments, US20080213899A1 by Connecticut university, US20110193510A1 by Newcastle Innovation, US5225750A and US5229679A by Prima Meat Packers, US20090069712A1 by Piezo Resonance Innovations, US5877008A by Lockheed Martin Energy Systems, and US20030059936A1 by Micronas GmbH. Consider the following three as being most relevant to the present invention.
US6251658B1 to BurleighInstruments, "inert impact drivers for cytological impact applications", discloses an Inertial impulse injection device that uses opposing piezoelectric or electrostrictive actuators to drive the movement of an Inertial mass and produce vibration of a micropipette or microelectrode. The flexure mechanism used in the present invention is not disclosed. The present invention also does not use two opposing piezoelectric actuators to generate the vibrations.
"rotanically inflating injector", US20080213899a1, discloses a syringe which penetrates an oocyte by rotating a micropipette with a rotary motor. The present invention does not involve rotating or using a rotary motor.
"positioning system and method", US20110193510a1, discloses a nanopositioning stage that uses piezoelectric actuators and a flexible mechanism for precise positioning. Although the present invention also uses piezoelectric actuators and flexible mechanisms, the device of the present invention is designed to produce vibrational motion through the transparent band rather than precise smooth motion positioning.
Disclosure of Invention
The device disclosed in the present application is a piezoelectric drill device capable of generating large axial vibrations and small lateral vibrations. The electric drill device can be used to effectively penetrate the Zona Pellucida (ZP) of oocytes and embryos with little distortion.
For the purpose, the invention adopts the following technical scheme:
in one aspect, the present invention provides a flexible guided piezoelectric drill device that generates large axial vibrations and small lateral vibrations for Zona Pellucida (ZP) penetration with small oocyte or embryo deformations.
Preferably, the electric drill device has a micropipette, a flexible member, a piezoelectric actuator, a flexible member holder, and a holding rod.
More preferably, the flexure includes a central portion, a plurality of flexure beams, an outer portion, and a flexure base.
Preferably, the flexure base is connected to the central portion.
Preferably, the flexible member is made of stainless steel by wire cut electrical discharge machining.
Preferably, the micropipette is fixed to the flexible member and is easily replaceable.
In a preferred embodiment, said central portion of the flexible member is connected to a conduit for providing negative pressure for oocyte or embryo aspiration, and said conduit is further connected to a pneumatic or hydraulic pump.
Preferably, a piezoelectric actuator is integrated on the flexure base.
Preferably, the piezoelectric actuator is secured against the base of the flexure and is preloaded with screws and metal shims.
Preferably, the outer portion of the flexible member is clamped by a flexible member holder and fastened by two screws.
Preferably, the flexure retainers are connected to the retaining rods.
Preferably, the holding bars are mounted on a micromanipulator having a motion stage to achieve precise positioning.
Preferably, the flexible beam connects the central portion and the outer portion of the flexible member by a double hinge.
Preferably, the hinge is an oval hinge.
A mechanical design method is also disclosed that uses a flexible member to guide movement of the micropipette in an axial direction.
In another aspect, a design of drive pulses is described. The designed drive signal is a pulse with the decaying resonance frequency of the micropipette and the flexible member.
Preferably, the drive pulses for driving the electric drill device are processed by filtering out the resonance frequency of the micropipette and the flexible member from the frequency spectrum of the drive pulses.
Drawings
One or more embodiments are described in detail below, by way of example, and with reference to the following figures, wherein:
fig. 1 illustrates a schematic diagram of a piezoelectric drill device.
Fig. 2 illustrates a top view of a piezoelectric drill device with a catheter assembly shown.
Figure 3 illustrates a schematic of a micropipette and its equivalent mechanics in contact with an oocyte or embryo.
Fig. 4 illustrates a schematic view of a flexible beam structure and its double oval hinge.
Fig. 5 illustrates filtered drive pulses for a piezoelectric actuator.
Fig. 6 illustrates the vibration of a micropipette mounted on a drill press apparatus measured by SEM.
Fig. 7 illustrates the axial vibration amplitude of the micropipette measured by the vibrating meter.
Fig. 8 illustrates ZP penetration with small oocyte deformation using an electric impact drill device.
Detailed Description
The present invention includes a mechanical design method of a piezoelectric drill unit generating large axial vibration and small lateral vibration, and a drive pulse design method for driving a piezoelectric actuator.
A. Structural design
As shown in fig. 1 and 2, the flexure 2 includes a central portion 3, a flexure beam 4, an outer portion 5, and a flexure base 6. The flexible guide design has a piezoelectric actuator 7 integrated on the flexure base 6. The movement of the micropipette 1 is guided by a plurality of flexible beams 4. The central portion 3 and the outer portion 5 are connected by a flexible beam 4 via an oval hinge. The flexure base 6 is connected to the central portion 3. The outer portion 5 is clamped by a flexible member holder 8. The flexure holders 8 are connected to holding bars 9, and the holding bars 9 are mounted on a micromanipulator (not shown).
The flexible member 2 may be made from a single piece of stainless steel by wire Electrical Discharge Machining (EDM). Thus, its overall structure is not affected by non-linearities caused by friction, wear and backlash.
The control of the actuation direction depends on the stiffness ratio between the transverse axes (Y and Z) and the axial axis (X). By achieving a low X-axis stiffness relative to the other two axes, the motion of the micropipette can be guided in the axial direction without concern for slight misalignments during assembly of the device.
A dynamic model of the micropipette is first established. The aim is to ensure that the lateral resonance frequency of the micropipette is avoided in the design of the flexible and the operating pulse sequence. Figure 3(a) shows a schematic of a micropipette in contact with an oocyte or embryo. Fig. 3(b) shows an equivalent mechanics system. The euler-bernoulli beam theory is used to derive the equation of motion of the micropipette tip, and the equation is solved by analysis to obtain the resonance frequency of the micropipette.
The power drill apparatus uses a flexible hinge guide mechanism. The flexible member uses elastic deformation of the material to transmit motion. For the flexure design, the inner portion of the flexure beam 4 is thickened to increase its out-of-plane (Z) stiffness, resulting in the double hinge beam structure shown in fig. 4. In contrast, circular hinges have high out-of-plane stiffness, but low range of motion. The lead angle hinge has a large range of motion, but poor out-of-plane stiffness. The oval hinge combines these characteristics to achieve a rigid flexure while maintaining relatively low stress at maximum extension of the flexure with long fatigue life, and therefore the oval hinge structure is chosen in the present design. For the selected flexible beam structure, the six parameters of the flexible beams to be determined are the number of the flexible beams and the thickness B of the flexible beamsTLength B of flexible beamLHinge shape, hinge pivot point thickness HTAnd hinge length HL。
Stainless steel was chosen for construction of the power drill apparatus because of its high yield strength, significantly higher than the predicted maximum stress encountered during vibration. The piezoelectric actuator 7 is integrated on the flexure base 6 and is preloaded with screws and metal shims. Sufficient pre-loading of the piezo is important to ensure its stability and avoid brittle fracture at operating frequencies near or above the resonant frequency of the flexure.
The X-axis resonance frequency must be higher than the frequency of the input pulses to avoid causing the flexure or micropipette to resonate. However, too high an X-axis resonant frequency requires high X-axis stiffness. The low stiffness ratio of the X-axis to the other axes can result in large off-axis motion and large lateral vibrations. An iterative finite element structure simulation is performed to determine the compliance parameter using the selected X-axis resonant frequency and preload.
B. Drive pulse design
The electric drill press device uses a pulse sequence to transfer ZP penetrating the oocyte or embryo with reduced energy, reducing oocyte damage. Forming a driving pulse sequence by applying a plurality of pulses with intervals therebetween per second; thus, it contains less energy than a continuous drive signal.
It is assumed that the fundamental frequency of the drive pulse is 18kHz and that the resonance frequencies of the micropipette and the flexure include 3-7kHz and above 20 kHz. A continuous sinusoidal signal with a frequency of 18kHz has all its spectral power at one point [ see fig. 5(a) (b) ]. Figure 5(c) shows a pulse with a fundamental frequency of 18 kHz. Its frequency response [ fig. 5(d) ] contains values from all frequencies that may cause transverse resonance, including 3-7kHz and above 20 kHz.
A band-stop filter with a cutoff frequency of [3kHz, 7kHz ] and a low-pass filter with a cutoff frequency of 20kHz were used to filter the original pulse signal shown in fig. 5 (c). Both filters are second order Infinite Impulse Response (IIR) filters.
The filtered pulse signal is shown in fig. 5(e) (f). It can be seen that the filtered pulse has a significantly attenuated 3-7kHz and an undesirable frequency range above 20 kHz.
When the piezo drill device is used for oocyte/embryo ZP penetration, the filtered pulse shown in fig. 5(e) is applied to the piezo actuator multiple times (e.g., 100 pulses) within one second, forming a pulse sequence as a driving signal for ZP penetration. The amplitude of the drive pulse can be varied by varying the peak voltage.
Example 1
Materials:
scanning electron microscopy (SEM, hitachi SU3500) was used to characterize the axial and lateral vibration amplitudes of the micropipette. The SEM-measured axial vibration amplitude of the micropipette was verified using a laser doppler vibrometer (OFV-5000, Polytec). The displacement resolution of the vibrating meter can reach 1 pm.
As a result:
SEM imaging has low bandwidth (20 Hz); therefore, when the micropipette is vibrated, its vibration envelope (vibrational envelope) appears as a blurred edge in SEM imaging [ see fig. 6(b) (C) ]. Measuring the distance between the edge of the micropipette and its corresponding vibration-induced blurred edge enables quantification of the axial and lateral vibration amplitudes of the micropipette.
Fig. 6(b) (c) correspond to 15kHz and 18kHz, respectively, as the frequency of the drive pulse supplied to the piezoelectric actuator. The peak voltage of the drive pulse was kept constant at 20V. When a 15kHz driving pulse [ fig. 6(b) ] is applied to the piezoelectric actuator, the micropipette tip has a lateral vibration amplitude of 500nm and an axial vibration amplitude of 1.2 μm. When the frequency of the drive pulse is increased to 18kHz [ fig. 6(c) ], the axial amplitude of vibration of the micropipette is still about 1.2 μm; however, the lateral vibration amplitude increased to 2 μm (0.5 μm relative to that produced by a 15kHz drive pulse). The axial vibration amplitude measured by a laser doppler vibrometer using 18kHz drive pulses was 1.2 μm [ see fig. 7], consistent with SEM measurements. In contrast, existing electric power drill devices all have large lateral vibration amplitudes (>20 μm) and extremely low axial vibration amplitudes (<0.1 μm).
Example 2
Materials:
mouse oocytes were collected from the Canadian Mouse Mutant library (Canadian Mouse Mutant replication) of the Toronto center, a Chinese for Phonogenics. Oocytes were observed using an inverted microscope (Nikon Ti, Nikon Microspecopes) and a CCD camera (acA1300-30gm, Basler).
As a result:
based on the test of 45 mouse oocytes, the ZP penetration success rate using the piezo-electric drill device of the present invention was 100%. Fig. 8(a) (b), corresponding to a supplied 18kHz drive pulse and 20V peak voltage, shows that the piezo drill device is able to penetrate the ZP of a mouse oocyte, with oocyte deformation as small as 3.4 μm. In contrast, existing electric drill devices produce mouse oocyte deformations of greater than 10 μm unless damped with a drop of mercury in a micropipette.
Claims (13)
1. A flexible guided piezo-electric drill device producing large axial and small lateral vibrations for oocyte or embryo Zona Pellucida (ZP) penetration with small oocyte or embryo deformation, said piezo-electric drill device having a micropipette, a flexible member, a piezo-electric actuator, a flexible member holder and a holding rod, said flexible member comprising a central portion, a plurality of flexible beams, an outer portion and a flexible member base, the micropipette tip having a lateral vibration amplitude of 500nm and an axial vibration amplitude of 1.2 μm when 15kHz drive pulses are applied to the piezo-electric actuator.
2. The device of claim 1, wherein the flexure base is connected to the central portion.
3. The device of claim 1, wherein the flexible member is formed from stainless steel by wire electrical discharge machining.
4. The device of claim 1, wherein the micropipette is secured to the flexible member and is easily replaceable.
5. The device according to claim 1, wherein said central portion of the flexible member is connected to a conduit for providing negative pressure for oocyte or embryo aspiration, and said conduit is further connected to a pneumatic or hydraulic pump.
6. The apparatus of claim 1, wherein the piezoelectric actuator is integrated on the flexure base.
7. The device of claim 6, wherein the piezoelectric actuator is secured against the flexure base and pre-loaded with screws and metal shims.
8. The device of claim 1, wherein the outer portion of the pliable component is clamped by a pliable component holder and secured by two screws.
9. The device of claim 1, wherein the pliable component holder is connected to the holding rod.
10. The apparatus of claim 1, wherein the holder bar is mounted on a micromanipulator having a motion stage to achieve precise positioning.
11. The apparatus of claim 3, wherein the compliant beam connects the central portion and the outer portion of the flexure via a double hinge.
12. The device of claim 11, wherein the hinge is an oval hinge.
13. The apparatus of claim 1, wherein the drive pulse for driving the electric drill apparatus is processed by filtering out the resonance frequency of the micropipette and the flexible member from the frequency spectrum of the drive pulse.
Applications Claiming Priority (3)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| US201762508361P | 2017-05-18 | 2017-05-18 | |
| US62/508,361 | 2017-05-18 | ||
| PCT/CN2018/087080 WO2018210266A1 (en) | 2017-05-18 | 2018-05-16 | Flexure-guided piezo drill with large axial vibration and small lateral vibration |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN110719952A CN110719952A (en) | 2020-01-21 |
| CN110719952B true CN110719952B (en) | 2022-04-05 |
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| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201880031544.0A Active CN110719952B (en) | 2017-05-18 | 2018-05-16 | Flexible guiding piezoelectric drill device with large axial vibration and small transverse vibration |
Country Status (3)
| Country | Link |
|---|---|
| CN (1) | CN110719952B (en) |
| CA (1) | CA3091527A1 (en) |
| WO (1) | WO2018210266A1 (en) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6251658B1 (en) * | 1999-06-18 | 2001-06-26 | Burleigh Instruments, Inc | Inertial impact drill for cytological applications |
| CN102245348A (en) * | 2008-10-09 | 2011-11-16 | 纽卡斯尔创新有限公司 | A positioning system and method |
Family Cites Families (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20030220644A1 (en) * | 2002-05-23 | 2003-11-27 | Thelen Sarah L. | Method and apparatus for reducing femoral fractures |
| US7728488B2 (en) * | 2006-05-25 | 2010-06-01 | Katholieke Universiteit Leuven | Positioning motor and apparatus |
| US20080213899A1 (en) * | 2006-10-12 | 2008-09-04 | University Of Connecticut | Rotationally Oscillating Injector |
-
2018
- 2018-05-16 WO PCT/CN2018/087080 patent/WO2018210266A1/en active Application Filing
- 2018-05-16 CA CA3091527A patent/CA3091527A1/en not_active Abandoned
- 2018-05-16 CN CN201880031544.0A patent/CN110719952B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US6251658B1 (en) * | 1999-06-18 | 2001-06-26 | Burleigh Instruments, Inc | Inertial impact drill for cytological applications |
| CN102245348A (en) * | 2008-10-09 | 2011-11-16 | 纽卡斯尔创新有限公司 | A positioning system and method |
Also Published As
| Publication number | Publication date |
|---|---|
| CA3091527A1 (en) | 2018-11-22 |
| WO2018210266A1 (en) | 2018-11-22 |
| CN110719952A (en) | 2020-01-21 |
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