Fluorescence In Situ Hybridization

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F r e q u e n t Q u e s t i on s A s k e d ab ou t F l u or e s c e n c e I n S i t u H yb r i d i z at i on F I S H Wh at i s F I S H F l uor e s c e nc e i n s i t u hy b r i di z a t i on F I S H m a ke s i t pos s i bl e f or t hos e bi ol ogi c a l r e s e a r c he r s t o vi s ua l i z e a nd m a p t he ge ne t i c m a t e r i a l i n a n i ndi vi dua l s c e l l s i nc l udi ng s pe c i f i c ge ne s or por t i ons of ge ne s . A nd i t i s a l s o us e d f or unde r s t a ndi ng a va r i e t y of c hr om os om a l a bnor m a l i t i e s a nd ot he r ge ne t i c m ut a t i ons . H ow d oe s F I S H w or k F I S H i s a us e f ul t ool f or i ns t a nc e f or he l p i ng a r e s e a r c he r or c l i ni c i a n t o i de nt i f y t he l oc a t i on of a pa r t i c ul a r ge ne f a l l s w i t hi n a n i ndi vi dua l s c hr om os om e s . T he f i r s t s t e p i s t o pr e pa r e s hor t s e que nc e s of s i ngl e - s t r a nde d D N A t ha t m a t c h a por t i on of t he ge ne w hi c h t he r e s e a r c he r i s l ooki ng f or . T he s e a r e c a l l e d pr obe s . N e xt i s t o l a be l t he s e pr obe s by a t t a c hi ng one of a num be r of c ol or s of f l uor e s c e nt dy e . W he n a pr obe bi nds t o a c hr om os om e i t s f l uor e s c e nt t a g pr ovi de s a w a y f or r e s e a r c he r s t o s e e i t s l oc a t i on. Wh at i s F I S H u s e d f or S c i e nt i s t s us e t hr e e di f f e r e nt t y p e s of F I S H pr obe s e a c h of w hi c h ha s a di f f e r e nt a ppl i c a t i on: L oc u s s p e c i f i c p r o b e s i s us e d t o bi nd t o a pa r t i c ul a r r e gi on of a c hr om os om e . T hi s t y p e of pr obe i s he l pf ul w he n r e s e a r c he r s ha ve i s ol a t e d a s m a l l por t i on of a ge ne a nd w a nt t o de t e r m i ne on w hi c h c hr om os om e t he ge ne i s l oc a t e d or how m a ny c opi e s of a ge ne e xi s t w i t hi n a pa r t i c ul a r ge nom e . A l p h oi d or c e n t r om e r i c r e p e at p r ob e s a r e t o de t e r m i ne w he t he r a n i ndi vi dua l ha s t he c or r e c t num be r of c hr om os om e s . T hi s t y p e of pr obe s a r e ge ne r a t e d f r om r e pe t i t i ve s e que nc e s f ound i n t he m i ddl e of e a c h c hr om os om e . R e s e a r c he r s a l s o us e t he s e pr obe s i n c om bi na t i on w i t h " l oc us s pe c i f i c pr obe s " t o de t e r m i ne w he t he r a n i ndi vi dua l i s m i s s i ng ge ne t i c m a t e r i a l f r om a pa r t i c ul a r c hr om os om e . Wh ol e c h r om os om e p r ob e s a r e a c t ua l l y c ol l e c t i ons of s m a l l e r pr obe s e a c h of w hi c h bi nds t o a di f f e r e nt s e que nc e a l ong t he l e ngt h of a gi ve n c hr om os om e . U s i ng m ul t i pl e pr obe s l a be l e d w i t h a m i xt ur e of di f f e r e nt f l uor e s c e nt dy e s s c i e nt i s t s a r e a bl e t o l a be l e a c h c hr om os om e i n i t s ow n uni que c ol or . T he r e s ul t i ng f ul l - c ol or m a p of t he c hr om os om e i s know n a s a s pe c t r a l ka r y o t y p e . W hol e c hr om os om e pr obe s a r e pa r t i c ul a r l y us e f ul f or e xa m i ni ng c hr om os om a l a bnor m a l i t i e s f or e xa m pl e w he n a pi e c e of one c hr om os om e i s a t t a c he d t o t he e nd of a not he r c hr om os om e .

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F or s uc h m a ny a pp l i c a t i ons F I S H ha s l a r ge l y be e n r e pl a c e d by t he us e of m i c r oa r r a y s . H ow e ve r F I S H r e m a i ns us e f ul f or s om e t e s t s . F I S H m a y a l s o be us e d t o s t udy c om pa r i s ons a m ong t he c hr om os om a l a r r a nge m e nt s of ge ne s a c r os s r e l a t e d s pe c i e s .

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