Volume 13, Issue 3 (11-1999)                   Med J Islam Repub Iran 1999 | Back to browse issues page

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ESMAILNEJAD MOGHADDAM A, KARIMPOUR MALEKSHAH A, HIROI M, SAITO H. INTRACYTOPLASMIC SPERM INJECTION (lCSI) IN MICE BY A NEW HOLDING PIPETTE. Med J Islam Repub Iran. 1999; 13 (3) :219-223
URL: http://mjiri.iums.ac.ir/article-1-940-en.html
From the Department of Anatomy and Embryology, Medical School, Mazandaran University of Medical Sciences, Sari, I R. Iran
Abstract:   (2086 Views)
Mice are ideal models for ICSI experiments because of the ease of culturing eggs/embryos in vitro and the availability of ample genetic information. Unfortunately, it has been extremely difficult. In this study we improved the mouse ICSI method by using a new holding pipette that was made of two pipettes such that one pipette was pulled and heat merged into the other one. The outer pipette had an outer diameter of 120 J..l. and an inner diameter of 80 J..l. to 85 J.1 (about mouse oocyte diameter). The inner pipette had an outer diameter of 80 µ to 85 µ. and an inner diameter of 55 µ to 60 µ. which was polished and narrowed to 20 J..l.on amicroforge. The distance between the tips of the two pipettes was adjusted to 120 µ. to 160 µ (1.5 to 2 fold longer than mouse oocyte diameter). Of 307 oocytes which were injected with a single spermatozoon, 206 (67.1 %) survived and 93 (45.1 %) of surviving oocytes showed normal fertilization (2 pronuclei and second polar body). Of 109 oocytes which were only sucked into the holding pipette (control group), 105 (96.3%) survived and only 4 (3.8%) of them became activated parthenogenically. U sing this new holding pipette, the oocyte is sucked into a glass tunnel and elongates to a reasonable length therefore the injection axis will be increased and piercing of the oolemma can be performed easily.
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