By Botros Rizk, Markus Montag
Compiled through specialists in Reproductive drugs, with contributions from the world over revered experts, this leading edge textual content shall we the total crew in Reproductive medication get actually at the comparable web page. Taking a cook-book method of the operational techniques within the laboratory and within the sanatorium, it info what should be ready upfront, what has to be ready previous a similar day, and what steps to take earlier than, in the course of, and after the method itself. this is often a vital instrument for making sure all employees - no matter if skilled or starters - could be convinced of their projects and are involved with what's anticipated of them and their colleagues.
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Extra info for Standard operational procedures in reproductive medicine : laboratory and clinical practice
D. Sakkas, E. Delpiano, S. Cayli, E. Ward, A. Revelli, and G. Huszar. 2005. ” Fertility and Sterility 84: 1665–73. E. Cognigni, S. Bernardi, E. Troilo, W. Ciampaglia, and M. Filicori. 2010. ” Fertility and Sterility 93: 598–604. , S. Eid, D. Woodhouse, M. Perloe, S. Smith, J. Witmyer, K. Ivani, C. D. Ball, T. Elliot, and J. Lieberman. 2013. ” Human Reproduction 28: 306–14. 10 IVF—Insemination Markus Montag Required Equipment • Stereo microscope with heated plate • Lamina flow hood with build-in heated plate [Optional: free-standing heating plate] • Incubator with appropriate % CO2/eventually low oxygen (5%–7% O2) • Pipette equipment for sperm pipetting (1–10 µL pipette + tips) • Gloves (non-powder) • Documentation chart To Be Prepared on the Day Before • Dish for COC culture with normal IVF culture media/with or without oil (gassed) Preparation on the Day • Isolate COC as described (Chapter 4).
Mayer, W. Arzt, and G. Tews. 2011. ” Reproductive Biomedicine Online 22: 37–43. M. A. Land. 2011. ” Human Reproduction Update 17: 719–33. , M. Maurer, O. Shebl, K. Dreier, G. Ziehr, G. Schappacher-Tilp, E. Petek, and T. Ebner. 2013. ” Reproductive Biomedicine Online 27: 81–8. World Health Organization. Laboratory Manual for the Examination and Processing of Human Semen. 5th ed. Geneva: WHO Press, 2010. 9 Sperm Selection Using Hyaluronan Binding Denny Sakkas Required Equipment • Stereo microscope with heated plate • Heating block for dishes • Pipette equipment for dispensing media, polyvinylpyrrolidone (PVP) and sperm sample (handling pipette/5–20 µL pipette + tips) • High-powered inverted microscope used for ICSI • Gloves (non-powder) • Labeling and recording documentation • PICSI sperm selection dishes (physiological intracytoplasmic sperm injection—PICSI) • Soluble hyaluronan solution Adjunct Protocols • Sperm preparation protocol • ICSI protocol Selection of Sperm • Identify patient sperm sample, egg dish after hyaluronidase treatment and patient documentation.
Viability Test Using Theophylline • Pipette a small volume of sperm preparation (<5 µL) into one 50 µL medium droplet. 5–5 µL theophylline ready to use sperm activation medium, mix gently and incubate at a warm plate for 5 min. • After 5 min screen droplet for spermatozoa with motility (flickering sperm tail). • Isolate motile spermatozoa with an injection capillary, aspirate head first. • Release spermatozoa in the small collection droplet. • Note: Theophylline activity will decrease after 20 to 30 min.