A new clinical non invasive method for live detection of mature M II Oocyte including Meiotic Spindle, ZP & Polar Body during ICSI to reduce ART failure
Iman Rezaei (1)
Scientific Dept manager of Iran Panam Co. No 12, 3rd Floors, Bldg. 108, Karimkhan Zand St, Tehran, Iran. Science@iran-panam.com
Now days, it is too significant to have a live check for cell & molecular condition of M II Oocyte during Intra Cytoplasm Sperm Injection (ICSI).
The visualizing of Meiotic Spindle to check the viability, polarity, size and situation of it, is valuable to select the mature oocyte for ICSI. In addition, place of Meiotic Spindle would be located at 12 or 6 o’clock. It is critical for oocyte and embryo to avoid any chromosomal abnormalities during ICSI in Assisted Reproductive Technologies (ART).
We are presenting the clinical use of the unique Cri Oosight Imaging System to visualize the Meiotic Spindle during ICSI & Nuclear Transfer.
Material & methods:
The Inverted Microscope with warm stage and RI Integra Ti Micromanipulator, Saturn Active Laser system with Cri Oosight Imaging System, including digital camera, LC Compensator, Cri Filter, Oosight Imaging Meta Software.
Oocyte were collected and checked & transfer to the glass dishes which were pre warmed in Incubators, and then locate on Inverted Microscope equipped by Cri Oosight Imaging System.
First, turn on the PC, locate the Cri Filter and LC Compensator and make sure the digital camera is on then do double click on software & observe the oocytes in Live image mode, adjust the brightness & then click on Blinking Oosight and finally click on Live Oosight mode & explore the Meiotic Spindle condition. Choose the right position for injection.
The chance of fertility will increase up to 20% and the chance of embryo development will improve up to 23% & in opposite the chance of genetic abnormalities will reduce.
It is a useful in use clinical system to make embryologist able to select a mature healthy oocytes for ICSI and prevent chance of genetic abnormalities and developmental failures.
Battaglia DE, Goodwin P, Klein NA, Soules MR (1996) Influence of maternal age on meiotic spindle assembly in oocytes from naturally cycling women. Hum Reprod 11(10), 2217 – 2222.
Chatzimeletiou K, Morrison EE, Prapas N, Prapas Y, Handyside AH (2005) Spindle abnormalities in normally developing and arrested human preimplantation embryos in vitro identified by confocal laser scanning microscopy. Hum Reprod 20(3), 672 – 682.
Chen CK, Wang CW, Tsai WJ, Hsieh LL, Wang HS, Soong YK (2004) Evaluation of meiotic spindles in thawed oocytes after vitrification using polarized light microscopy. Fertil Steril 82 (3), 666 – 672.
Keefe D, Tran P, Pellegrini C, Oldenbourg R (1997) Polarized light microscopy and digital image processing identify a multilaminar structure of the hamster zona pellucida. Hum Reprod 12(6), 1250 – 1252. 1. 2. 3. 4.
Kilani SS, Cooke S, Kan AK (2006) Chapman MG. Do age and extended culture affect the architecture of the zona pellucida of human oocytes and embryos? Zygote 14 (1), 39 – 44.
Moon JH, Hyun CS, Lee SW, Son WY, Yoon SH, Lim JH (2003) Visualization of the metaphase II meiotic spindle in living human oocytes using the Polscope enables the prediction of embryonic developmental competence after ICSI. Hum Reprod 18 (4), 817 – 820.
Nottola SA, Macchiarelli G, Coticchio G, Bianchi S, Cecconi S, De Santis L, Scaravelli G, Glamigni C and Borini A (2007) Ultrastructure of human mature oocytes after slow cooling cryopreservation using different sucrose concentrations. Hum Reprod 22 (4), 1123 – 1133.
Noyes, H. C. Chang, H. Liu, P. Labella, L. Meng, J. A. Grifo (2006) Presence of meiotic spindle predicts embryo competence following oocyte cryopreservation. Fertil Steril 86 (2) S64 – S65.
Pelletier C, Keefe DL, Trimarchi JR (2004) Noninvasive polarized light microscopy quantitatively distinguishes the multilaminar structure of the zona pellucida of living human eggs and embryos. Fertil Steril 81 Suppl 1, 850 – 856.
Rienzi L, Martinez F, Ubaldi F, Minasi MG, Iacobelli M, Tesarik J and Greco E (2004) Relationship between meiotic spindle location with regard to the polar body position and oocyte developmental potential after ICSI. Hum Reprod 18 (6), 1289 – 1293.
Shen Y, Stalf T, Mehnert C, De Santis L, Cino I, Tinneberg HR, Eichenlaub-Ritter U (2006) Light retardance by human oocyte spindle is positively related to pronuclear score after ICSI. Reprod Biomed Online 12 (6), 737 – 757.
Shen Y, Betzendahl I, Sun F, Tinneberg HR, Eichenlaub-Ritter U (2005) Non-invasive method to assess genotoxicity of nocodazole interfering with spindle formation in mammalian oocytes. Reprod Toxicol 19 (4), 459 – 471.
Wang WH, Meng L, Hackett RJ, Oldenbourg R, Keefe DL (2002) Rigorous thermal control during intracytoplasmic sperm injection stabilizes the meiotic spindle and improves fertilization and pregnancy rates. Fertil Steril 77 (6), 1274 – 1277.