WORKSHOP BOOK Fluorescent in Situ Hybridisation - FISH - on Human Sperm

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Scor<strong>in</strong>g - Count<strong>in</strong>g Fields should be found at 10X or 20X magnification. Then, magnification should be <strong>in</strong>creased to 40X and gradually to 100X with oil, to count the spots. There should be only one spot of each chromosome <strong>in</strong> every sperm cell. (Chromosomes 18, X and 13at our experiment emit green colour, whereas chromosomes Y and 21 emit red colour). TIP: •Spot count<strong>in</strong>g should be careful. The marg<strong>in</strong>s of the cells should be clear <strong>in</strong> order to know that the spots come from chromosomes. •2 spots <strong>in</strong> a cell should be clearly separated and have the same <strong>in</strong>tensity <strong>in</strong> order to be considered as aneuploid. (Blanco et al, 1996) Hands-on Workshops, Thessaloniki, 30.08 – 5.09 2014 9 Sperm <strong>FISH</strong> QC Use always your control Hands-on Workshops, Thessaloniki, 30.08 – 5.09 2014 10
Sperm <strong>FISH</strong> External QC Hands-on Workshops, Thessaloniki, 30.08 – 5.09 2014 11 Sperm <strong>FISH</strong> External QC Hands-on Workshops, Thessaloniki, 30.08 – 5.09 2014 12
Page 1 and 2:
1 WORKSHOP <strong
Page 3 and 4:
3 Content Workshop Book Embryolab A
Page 5 and 6:
5 Hands-on Workshop 1 Fluor
Page 7 and 8:
7 Programme 8:30 - 8:45 Pick- up fr
Page 9 and 10: Aneuploidy in male
Page 11 and 12: Spermatogenesis Hands-on Workshops,
Page 13 and 14: Paternally derived aneuploidies A 2
Page 15 and 16: Reciprocal Translocation The most f
Page 17 and 18: Interchromosomal Effect (ICE) Aneup
Page 19 and 20: Aneuploidy in male
Page 21 and 22: Sperm morphology Data regard<strong
Page 23 and 24: Infertile males (46,XY) Cryptorhidi
Page 25 and 26: DNA Fragmentation Index (DFI) Sever
Page 27 and 28: Occupational/Life style hazards Stu
Page 29 and 30: 41 Mechanisms again</strong
Page 31 and 32: Take home message At present, PGD i
Page 33 and 34: In summary • Multicolour
Page 35 and 36: FISH on Spermatozo
Page 37 and 38: FISH: What can it
Page 39 and 40: A DNA probe finds
Page 41 and 42: Hybridisation targ
Page 43 and 44: Sperm-FISH protoco
Page 45 and 46: 1. Preparation of sample Nuclear De
Page 47 and 48: 2. Co-Denaturation (target DNA & pr
Page 49 and 50: 5. Examine slides
Page 51 and 52: FISH Advantages )
Page 53 and 54: Figure 5. Model of chromosome organ
Page 55 and 56: 55 FISH technique:
Page 57 and 58: Tip for spreading
Page 59: Post Hybridization Wash The aim of
Page 63 and 64: Issue Date: 05/12/2012 Version: 1 R
Page 71 and 72: Our generous Sponsors 71
Page 73 and 74: 73 Certificate of attendance A Cert
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WORKSHOP BOOK Fluorescent in Situ Hybridisation - FISH - on Human Sperm

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