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XYClone is for non-clinical research use only in USA. hESC research using the XYClone should be conducted in accordance with any applicable Federal, State or local laws and regulations.
Please Note: The XYClone™ is for use ONLY with ANIMAL and STEM CELL APPLICATIONS. (For Clinical Laser Assisted Hatching and Biopsy applications, please see the ZILOS-tk.)
XYClone for Laser-assisted ICSI
With the success of laser-assisted ICSI for assisted reproduction in humans, the possibilty of laser-assisted ICSI in other species is being explored as well. Please note that while we present two mouse ICSI posters below, laser-assisted ICSI protocols in the mouse are in their infancy and further study and optimization is required.
Laser-ICSI: A Novel Method For Performing Mouse ICSI That Avoids Piezo-drilling (Download full poster)
Background
- Conventional ICSI has a low survival rate in mouse oocytes due to the high level of lysis.
- The method of injection has therefore been adapted to use of a piezo-drill to breach the fragile oolemma and thereby minimise oocyte stress and increase survival.
- Fragile oocytes' have also been reported in the human, where a strategy of lasering a hole into the zona pellucida (ZP) immediately prior to conventional ICSI has been shown to improve survival.
- Since membranes consist of phospholipid bilayers, we hypothesised that a laser could also be used to ‘melt' a small area of the oolemma to permit insertion of an injection pipette for ICSI without damaging the oocyte.
- This novel technique may provide an alternative method for mouse ICSI that minimises oolemma trauma and avoids the use of a piezo-drill.
Objective
- To investigate whether mouse ICSI could be successfully performed using the XYClone laser (Hamilton Thorne, Beverly, USA).
Conclusions
- This preliminary experiment demonstrates a novel technique for injecting spermatozoa heads into oocytes using an infrared laser rather than piezo-drill.
- Laser-ICSI may provide an alternative method of producing viable mouse embryos. A comparison is warranted between ICSI
performed using a laser and a piezo-drill.
XYClone™ Laser System: A Novel Technique for Mouse Intracytoplasmic Sperm Injection (ICSI) (Download full poster)
William A. Richie, Embryologist, Division of Gene Expression and Development, Roslin Institute, Roslin Midlothian UK EH25 9PS and K. Bradley, Hamilton Thorne, Beverly, MA 01915 USA
Introduction
A piezo actuated device has long been recognized as a necessary tool for the production of transgenic mice via intracytoplasmic sperm injection. Problems associated with the time required to learn how to use a Piezo system efficiently, as well as the additional cost of purchasing pipettes and the handling and disposal of mercury are eliminated when using the XYClone laser system.
Goal
The purpose of this study was to investigate the efficiency of injection and development of mouse oocytes to blastocyst when the XYClone laser was utilized for membrane breaching instead of a piezo impact driver.
Conclusions
- The XYClone laser system is an easy and effective way to breach the zona pellucida and oolemma membranes during the course of mouse ICSI.
- There is a high percentage of survival as well as an excellent rate of progression to blastocyst with the difficult B6D2F1 model.
- The XYClone laser system is extremely easy to use, greatly reducing the learning curve associated with the piezo drill technique.
- There were no hazardous chemicals to handle or dispose of and no need to purchase expensive injection pipettes.
Laser is fired 3 times to cut a channel in the zona pellucida. 
Sperm had (in red oval) is injected into cytoplasm.
Video of Mouse ICSI
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WINDOWS MEDIA |
Mouse ICSI
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