Hi ORO experts,
I'm struggling with ORO background precipitates in arterial tissues. The protocol I use works very well on human atherosclerotic plaques which are full of lipids and I don't notice the background, but when it's a healthy artery with only droplets of lipids I see lots of precipitates from ORO solution. I use ready to to use 0.5% ORO/isopropanol solution from Sigma which I dilute in water (60%), leave for 10min and filter (tried all sort of filters: 0.22um, o.45um, whateman paper and a combination of whateman paper/0.22um filter). Tissues are then immersed in 10% neutral buffered formalin for 5 min, then 5 min in 60% isopropanol, ORO was added to all tissues (positive control is the human plaque) and incubated for 10min. Slides are then immersed quickly in 60% isopropanol then wash in water before I counter stain with haematoxylin. everything works except the precipitates. I tried warming but the same issueup the solution in a heat block @ 60C for 2 hours as well . Any idea how to fix this please? thanks in advance!
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