Hello all,
I am working with mouse organs infected with Trypanosoma cruzi parasites. When animals are killed they are in the chronic stage of the disease, and their organs are extracted in Category 3 facility, where they are fixed with a solution of 4% PFA for 24 hours. Then they need to be further processed and I was wondering if that could be safely done outside of Category 3 facility.
I know I could technically take the organs, fix them and culture them but considering the 'stubbornness' of these parasites I should probably wait few weeks/months to make sure that they are definitely dead and the sample is no more infectious.
So I was wondering if there is already a proof in the literature that the fixation kills the parasites in the tissues? And if so, is it then safe to take the organs outside of Categority 3 facility and process them in the regular lab space?
Any help will be much appreciated.
To anyone that is still interested in this issue.
I took heavily infected mouse brain and heart (the infection was confirmed with in vivo imaging of bioluminescence) and fixed them overnight in 4% of chilled PFA. I then manually broke down the organs into homogenates and cultured them in the media that I usually use for trypomastigotes culture. Unfortunately, the brain sample got contaminated around day 24, but heart sample was cultured for the overall of 70 days. No motile trypomastigotes were seen throughout the entire experiment which confirms that PFA fixation of intact organs indeed kills the parasites and infected samples can be processed outside of Category 3 facility.
Kind regards.