Target spots on microarray - (Sep/16/2005 )
Hello,
I've been experiencing reverse donut spots lately on my microarrays (i.e. bright center, faded middle ring, brighter outside ring) and am at a loss as to what is causing this morphology.
At my old lab, we bought pre-coated slides and we never had a problem with this type of spot. Here at my new lab, we coat the glass slides in-house using a poly-lysine coating. I've found the microarrays used the day they are printed are great - no funny spots. However, even a week after their initial printing, target spots start appearing after scanning. Is this a coating problem? A rehydration problem? Any advice or shared experiences are most welcome.
Thanks!
Eva
-evatam-
QUOTE (evatam @ Sep 16 2005, 11:17 AM)
Hello,
I've been experiencing reverse donut spots lately on my microarrays (i.e. bright center, faded middle ring, brighter outside ring) and am at a loss as to what is causing this morphology.
At my old lab, we bought pre-coated slides and we never had a problem with this type of spot. Here at my new lab, we coat the glass slides in-house using a poly-lysine coating. I've found the microarrays used the day they are printed are great - no funny spots. However, even a week after their initial printing, target spots start appearing after scanning. Is this a coating problem? A rehydration problem? Any advice or shared experiences are most welcome.
Thanks!
Eva
I've been experiencing reverse donut spots lately on my microarrays (i.e. bright center, faded middle ring, brighter outside ring) and am at a loss as to what is causing this morphology.
At my old lab, we bought pre-coated slides and we never had a problem with this type of spot. Here at my new lab, we coat the glass slides in-house using a poly-lysine coating. I've found the microarrays used the day they are printed are great - no funny spots. However, even a week after their initial printing, target spots start appearing after scanning. Is this a coating problem? A rehydration problem? Any advice or shared experiences are most welcome.
Thanks!
Eva
hi eva,
in my opinion donuts are quite common if the intensity of the light is very high in the spots(means high concentrations of cyt3 or cyt5). i am not a expert in physics of light, but i understood that it is based on the phenomenon of light.
u can try to avoiding this problem by reducing the quantity of oligonucleotides you are printing on slide or by using low concentrations of target cDNA or RNA which is labelled.
gud luck.
-payeli-