I would wash after the lithium carbonate.Is your eosin alcoholic? I would make it in 70% alcohol. Then dip slides in 80% alcohol between the lithium and  eosin. Then go directly into 95% alcohol, absolute, xylene as usual.Joyce________________________________From: histonet-bounces@lists.utsouthwestern.edu on behalf of Ian WardSent: Tue 12/7/2004 4:24 AMTo: histonet@lists.utsouthwestern.eduSubject: [Histonet] H and E stainingHi allI could use some advice, I have been trying to H and E stain some mouse sections of the lungs, brain, spleen, liver, kidney, pancreas and heart. i have used the following method:-Xlylene     5minutesXlylene     5minutes100% IMS 5minutes100% IMS 5minutes90% IMS  5minutes70% IMS  5minutes50% IMS  5minutesrunning tap water   5minutesFiltered Haematoxylin   5minutesQuick wash in running tap waterDip 20 times in Acid AlcoholQuick wash in running tap waterFew seconds in Lithium Carbonate1% Eosin   5minutesQuick rince in running tap waterTake quickly though alcohols to xylene againMountMy problem is that for some reason very little of the Eosin can be seen on the sections when I come to view them. Any ideas?CheersIanIan WardSenior TechnicianE FloorThe Medical SchoolQMCNottinghamNG7 2UHTel: 0115 9249924 Ext 36179This message has been scanned but we cannot guarantee that it and anyattachments are free from viruses or other damaging content: you areadvised to perform your own checks.  Email communications with theUniversity of Nottingham may be monitored as permitted by UK legislation._______________________________________________Histonet mailing listHistonet@lists.utsouthwestern.eduhttp://lists.utsouthwestern.edu/mailman/listinfo/histonetConfidentiality Notice ** The information contained in this message may be privileged and is confidential information intended for the use of the addressee listed above. If you are neither the intended recipient nor the employee or agent responsible for delivering this message to the intended recipient, you are hereby notified that any disclosure, copying, distribution or the taking of any action in reliance on the contents of this information is strictly prohibited. If you have received this communication in error, please notify us immediately by replying to the message and deleting it from your computer.Thank you. Saint Josephs Health System, Inc.