WORKSHOP BOOK Fluorescent in Situ Hybridisation - FISH - on Human Sperm
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Scor<str<strong>on</strong>g>in</str<strong>on</strong>g>g - Count<str<strong>on</strong>g>in</str<strong>on</strong>g>g<br />
Fields should be found at 10X or 20X magnificati<strong>on</strong>. Then, magnificati<strong>on</strong> should<br />
be <str<strong>on</strong>g>in</str<strong>on</strong>g>creased to 40X and gradually to 100X with oil, to count the spots.<br />
There should be <strong>on</strong>ly <strong>on</strong>e spot of each chromosome <str<strong>on</strong>g>in</str<strong>on</strong>g> every sperm cell.<br />
(Chromosomes 18, X and 13at our experiment emit green colour, whereas<br />
chromosomes Y and 21 emit red colour).<br />
TIP:<br />
•Spot count<str<strong>on</strong>g>in</str<strong>on</strong>g>g should be careful. The marg<str<strong>on</strong>g>in</str<strong>on</strong>g>s of the cells should be clear <str<strong>on</strong>g>in</str<strong>on</strong>g><br />
order to know that the spots come from chromosomes.<br />
•2 spots <str<strong>on</strong>g>in</str<strong>on</strong>g> a cell should be clearly separated and have the same <str<strong>on</strong>g>in</str<strong>on</strong>g>tensity <str<strong>on</strong>g>in</str<strong>on</strong>g><br />
order to be c<strong>on</strong>sidered as aneuploid. (Blanco et al, 1996)<br />
Hands-<strong>on</strong> Workshops, Thessal<strong>on</strong>iki, 30.08 – 5.09 2014 9<br />
<strong>Sperm</strong> <str<strong>on</strong>g>FISH</str<strong>on</strong>g><br />
QC<br />
Use always your c<strong>on</strong>trol<br />
Hands-<strong>on</strong> Workshops, Thessal<strong>on</strong>iki, 30.08 – 5.09 2014 10
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