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For the preparation of density gradients
PureSperm 100 is a sterile colloidal silica suspension in an isotonic salt solution. It is optimized for the preparation of density gradients used to separate and purify human sperm from semen for use in Assisted Reproduction Technologies (ART). This system effectively separates normal sperm from lymphocytes, epithelial cells, abnormal, immature and senescent sperm, cell debris, bacteria and seminal fluid. Watch video tutorial
PureSperm 100 stock suspension should be diluted with PureSperm Buffer to provide layers of different densities required for the gradient. For optimal results, the sperm pellet should be washed subsequently in PureSperm Wash. These three products together form an optimized system for the preparation of sperm for ART.
Prepare two PureSperm gradients for each semen sample. This reduces the risk of overloading a single gradient, provides security when handling tubes or recovering sperm pellets, and provides two tubes to balance the centrifuge rotor.
It can be extremely difficult to obtain a high yield from motile sperm from highly viscous semen samples. Nidacon has developed a method that could be helpful to you in the lab. You simply add PureSperm Buffer to the ejaculate, incubate at 37°C for 15-30 minutes and the sample is ready to use. This will give you a much higher proportion of motile sperm.
Protocol:
For sperm preparation from a viscous semen sample:
1. Bring all solutions to room temperature (17 to 27°C).
2. Measure the volume of the semen sample.
3. Dilute 1+2 , 1 part PureSperm Buffer and 2 parts semen sample
(e.g. 0.5 ml PureSperm Buffer + 1.0 ml semen sample).
4. Incubate at 37°C for 15-30 minutes.
5. Mix using a pipette.
6. Ready for sperm preparation on a density gradient.
Article No. / Name / Volume
PS100-100 / PureSperm 100 / 100 mL
PS100-250/PureSperm 100 / 250 mL
PS100-1000K/PureSperm 100 / 4×250 mL
Stefania Luppi1, Monica Martinelli1, Elisa Giacomini2, Elena Giolo1, Gabriella Zito2, Rodolfo C Garcia3† and Giuseppe Ricci12*†
Reproductive Biology and Endocrinology 2015, 13:36
Iraj Amiri, Marzieh Ghorbani, Safora Heshmati
Journal of Clinical and Diagnostic Research. 2012 November, Vol-6(9): 1451-1453
Charoenchai Chiamchanya, Nattpawit Kaewnoonual, Pachara Visutakul, Sirikul Manochantr and Jirattikan Chaiya.
Asian J Androl. 2010 Mar;12:271-277.
Sakkas D, Manicardi GC, Tomlinson M, Mandrioli M, Bizzaro D, Bianchi PG, Bianchi
U.Hum Reprod. 2000 May;15(5):1112-6.
Henkel R, Kierspel E, Hajimohammad M, Stalf T, Hoogendijk C, Mehnert C, Menkveld R, Schill WB, Kruger TF.
Reprod Biomed Online. 2003 Oct-Nov;7(4):477-84.
P Ranganathan, A Agarwal
Fertil Steril, 2001
Allamaneni SS, Agarwal A, Rama S, Ranganathan P, Sharma RK.
Asian J Androl. 2005 Mar;7(1):86-92.
C.M. Nicholson L. Abramsson, S.E. Holm and E. Bjurulf
Human Reproduction, Vol. 15, No. 3, 662-666, March 2000
M. Tomlinson et al
Abstract Andrology in the nineties 1999
Agarwal, A. and Ranganathan, P.ESHRE 2001
J. A. Roudebush, et al.
Supplement of the 27th Annual Meeting of American Society of Andrology, 81 Seattle, USA (March/April 2002)
Nathalie Mousset-Siméon, Nathalie Rives, Lydie Masse, Florence Chevallier and Bertrand Mace
Journal of Andrology, Vol. 25, No. 6, November/December 2004