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Microinjection of FITC-dextran into mouse blastomeres to assess topical effects of zona photoablation
- Leyi Li, Santiago Munne, Frederick Licciardi, Joseph Neev, Yona Tadir, Michael Berns, Robert Godke, Jacques Cohen
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The objective of the current experiments was to investigate whether all or only some blastomeres from precompacted mouse embryos were affected by zona photoablation. The microbeam of xenon chloride excimer laser (308 nm) was guided through an inverted microscope (non-contact system). Topical effects of lasing were determined by microinjection of a vital fluorescent dye of high molecular weight (fluorescein isothiocyanate [FITC] dextran) into the cell immediately adjacent to the site of zona photoablation. This dye is only passed onto daughter blastomeres and therefore allows study of specific cell lines. Embryonic growth was assessed following cell separation at the morula and blastocyst stage. Four-cell embryos treated with the laser had significantly fewer cells 12 h after zona photoablation than control embryos. A similar effect was noted after 24 h between dye injected embryos and those injected and lased simultaneously, indicating potential toxic effects of the laser treatment on the embryo. Effects on the blastomere closest to the site of ablation were evaluated by calculating the ratio of dyed cells to the total number of cells at specific time intervals. The ratios were similar in the dye and laser + dye groups of treated 4-cell embryos 36 h after treatment (0.22 and 0.23, respectively), indicating that the dye was still present in approximately 25% of the cells and that the negative effect of photoablation was evenly distributed among the blastomeres. It is concluded that zona photoablation may have long-term detrimental effects of a non-topical nature on precompacted mouse embryos in spite of the apparent precision of the laser spot size.
Forewords
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- By Thomas M. Krummel, Professor and Chair Department of Surgery Stanford University School of Medicine Susan B. Ford Surgeon in Chief Lucile Packard Children's Hospital Palo Alto, CA, Linda C. Giudice, The Robert B. Jaffe Professor and Chair Department of Obstetrics, Gynecology and Reproductive Sciences University of California, San Francisco School of Medicine San Francisco, CA, Carmel J. Cohen, Professor of Clinical Obstetrics and Gynecology Division of Gynecologic Oncology Department of Obstetrics and Gynecology Columbia University Medical Center New York, NY, Jonathan S. Berek, Professor and Chair Department of Obstetrics and Gynecology Stanford University School of Medicine Palo Alto, CA, Alan H. DeCherney, Professor and Chairman Department of Obstetrics and Gynecology UCLA School of Medicine Los Angeles, CA, H. C. Mult Kurt Semm, Alan H. DeCherney, Louis E. Phaneuf Professor and Chairman Department of Obstetrics and Gynecology Tufts University Boston, MA, Yona Tadir, Department of Surgery Beckman Laser Institute & Medical Clinic Irvine, CA
- Camran Nezhat, Stanford University School of Medicine, California, Farr Nezhat, Mount Sinai School of Medicine, New York, Ceana Nezhat
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- Book:
- Nezhat's Operative Gynecologic Laparoscopy and Hysteroscopy
- Published online:
- 23 December 2009
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- 07 July 2008, pp xiii-xx
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Summary
Progress in surgical science has been characterized by a continuous cycle of innovation from bedside to bench and back to bedside. Beginning 30,000 years ago with the first bone needles to the current armamentarium today, each quantum leap has resulted from the convergence of technical advances and creative surgeons.
Some surgical capability has been enhanced by relatively simple or more complex tool manufacture or modification, usually for a single purpose. Kocher's addition of a tooth to a straight clamp facilitated the grasping of a thyroid goiter; the more modern fixed-ring retractors have added considerable utility in abdominal retraction.
A few very special tools or techniques revolutionize our work. The development of the simple balloon catheter by Fogarty was the seminal event in initiating the concept of all endovascular procedures, beginning with the procedure of intra-luminal thrombectomy. It has expanded to balloon dilatation, angioplasty, stent placement, and now drug delivery systems in the form of drug-eluting stents.
Dr. Camran Nezhat's creative and ingenious contribution to the field of laparoscopic surgery has been similarly revolutionary. Operating off the video monitor during endoscopic surgery by the addition of a video camera to the laparoscope as developed by Camran Nezhat was a critical step in facilitating the entire field of minimal access surgery, moving it out of its initial realm in gynecologic and pelvic surgery to the entire abdomen, the chest, and beyond.