WORKSHOP BOOK Fluorescent in Situ Hybridisation - FISH - on Human Sperm
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Issue Date:<br />
05/12/2012<br />
Versi<strong>on</strong>:<br />
1<br />
Rev.:<br />
0<br />
SOP-LAB-01: <str<strong>on</strong>g>FISH</str<strong>on</strong>g> ON HUMAN SPERM<br />
Page #:<br />
4 out of 7<br />
Prepared by: Glykeria Samolada Edited by: Vassilis Katsares Approved by: Alexia Chatziparasidou<br />
1. Purpose<br />
The purpose of this procedure is to describe the technique of <str<strong>on</strong>g>FISH</str<strong>on</strong>g> <strong>on</strong> human spermatozoa.<br />
2. Scope/Field of Applicati<strong>on</strong><br />
This procedure is taken place <str<strong>on</strong>g>in</str<strong>on</strong>g> the IVF lab and the genetic material to be exam<str<strong>on</strong>g>in</str<strong>on</strong>g>ed is isolated from human<br />
spermatozoa.<br />
3. Def<str<strong>on</strong>g>in</str<strong>on</strong>g>iti<strong>on</strong>s and Acr<strong>on</strong>yms<br />
IVF: In Vitro Fertilizati<strong>on</strong><br />
PBS: Phosphate Buffer Sal<str<strong>on</strong>g>in</str<strong>on</strong>g>e<br />
SSC: Sodium chloride – Sodium Citrate<br />
4. Resp<strong>on</strong>sibilities<br />
The current procedure is performed <strong>on</strong>ly by properly tra<str<strong>on</strong>g>in</str<strong>on</strong>g>ed pers<strong>on</strong>nel (geneticists and technicians), who are well<br />
experienced <str<strong>on</strong>g>in</str<strong>on</strong>g> sperm <str<strong>on</strong>g>FISH</str<strong>on</strong>g>. The IVF unit that provides this service, as well as the collaborated laboratories for the<br />
genetic analysis, must be certified accord<str<strong>on</strong>g>in</str<strong>on</strong>g>g to ISO 9001 and should be able to provide patients with genetic counsel<str<strong>on</strong>g>in</str<strong>on</strong>g>g<br />
and all necessary c<strong>on</strong>sent forms with clear <str<strong>on</strong>g>in</str<strong>on</strong>g>formati<strong>on</strong> <strong>on</strong> all possible outcomes and results.<br />
5. Materials Required<br />
Ma<str<strong>on</strong>g>in</str<strong>on</strong>g>tenance-c<strong>on</strong>sumables-c<strong>on</strong>sent forms:<br />
Prior to this procedure all equipment to be used should be validated and ma<str<strong>on</strong>g>in</str<strong>on</strong>g>ta<str<strong>on</strong>g>in</str<strong>on</strong>g>ed accord<str<strong>on</strong>g>in</str<strong>on</strong>g>g to the manufacturer’s<br />
<str<strong>on</strong>g>in</str<strong>on</strong>g>structi<strong>on</strong>s. All c<strong>on</strong>sumables, plastic ware and media used, should be CE marked and <str<strong>on</strong>g>in</str<strong>on</strong>g>dividually packaged or<br />
aliquoted, when possible. All c<strong>on</strong>sent forms c<strong>on</strong>cern<str<strong>on</strong>g>in</str<strong>on</strong>g>g the patients counsel<str<strong>on</strong>g>in</str<strong>on</strong>g>g, the procedure, the diagnosis and the<br />
possible outcomes should be clearly stated and signed by the patient(s).<br />
Hardware:<br />
Inverted microscope, <str<strong>on</strong>g>Fluorescent</str<strong>on</strong>g> microscope with filters, light source, hot plate, centrifuge.<br />
<strong>Sperm</strong> /cell manipulati<strong>on</strong><br />
2-20 µl micropipettes, 20-200 µl micropipettes, filtered tips (20 µl and 200 µl),<br />
Miscellaneous:<br />
Laboratory footwear and outfit, masks (Hospital and Home care Surgical Face masks), s<str<strong>on</strong>g>in</str<strong>on</strong>g>gle use robes, laboratory caps,<br />
sterile surgical gloves (latex, powder free).<br />
SOP-LAB-02: <str<strong>on</strong>g>FISH</str<strong>on</strong>g> ON HUMAN SPERM/01/00/05-12-2012
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