Re: [Histonet] fatty breast processing

From:Rene J Buesa

First of all, more important than the actual protocol is the fixation of the breast tissue, and in order to get good fixation, you have to start with THIN slices of breast, 3 mm thick OR LESS if at all possible.
After this thickness factor is obtained you will need a good fixation time. Lets say that you get the tissues  between 4&5 pm; the should be described preferentially, meaning, immediately and put in cassettes into the NBF at 5 or 6 PM. If you have a VIP and your end time (I gather that you mean the end time for the processing protocol) is at 10AM, you could start processing at midnight which will mean that the tissues will be in the retort in formalin from 5 or 6 PM until midnight and IF the sections are THIN enough, they will be fixed by then.
Start your ptotocol in the first alcohol station, with the following sequence:
 60%EthOlX50 min; 80%EthOlX50 min; 95%EthOlX50 min; 100%EthOlX1 hour;100%EthOlX45 min; 100%EthOlX45 in; XyleneX45 min; XyleneX45 min; Paraf.1X45 min; Paraf2 X50 min; Paraf 3X 55 min and Paraf 4 X1hour  This protocol will give 10 hours (without secondary fixation since it will skip station 2 and the tissues will be in the retort with the formalin from station 1).
This protocol will give you a (clearing+infiltration)/dehydration ratio of 0.97 which is a good ratio to avoid excessive dryness.
Really we stop having problems with breast (and any other difficult tissue) when we abandoned xylene and start using mineral oil.
Hope this will help you!
Rene J.

Dennis Gianotti  wrote:
We have a sakura VIP processor and need to process breast tissue that 
comes in between 4 & 5 pm and have it out the next day. What would be a 
good protocal to use on the processor using formalin, zylene and 
alcohols. The end time could be as late as 10 am the next day.

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