Efficient grinding of Drosophila heads for epigenomic analysis in small volumes
Experiments involving molecular analysis of rare cell types from Drosophila brains often requires large numbers of flies to recover enough material. This limits the experiments that can be done, since more specific targeting of neurons means smaller numbers of relevant tissue, and that targeted tissue is hard to isolate.
The grinder described here is optimized to miniaturize homogenization volumes and reduce sample loss. Combining this design with some methodological tweaks described in mini-INTACT protocol users can get ~100 fold reduction in required sample for downstream biochemical analysis.
This motorized grinder called ‘mini-INTACT homogenizer’ for tissue homogenization is optimized for extractions of neuronal nuclei from small volumes and small numbers of flies. mini-INTACT protocol combined with this homogenizer helps reduce the amount of fly heads needed for experiments about 100-fold when compared with previously described INTACT method in Drosophila (Henry et al., 2012) that used Yamato continuous flow homogenizer (US patent 4307846). Patent Link: https://patents.google.com/patent/US4307846A/
The mini-INTACT homogenizer was utilized in a study that investigated epigenetic changes caused by social isolation, with only 200-250 fly heads, targeting ~0.1% of fly neuronal nuclei. The new method, called mini-INTACT, opens the field of behavioral epigenetics in Drosophila to cell type-specific studies.
Link to mini-INTACT method: http://dx.doi.org/10.1186/s12915-019-0646-4
Opportunity: Free to make for Non-Profit Research by downloading the design here. See included hardware license.
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To cite the designs, please use DOI: https://doi.org/10.25378/janelia.32326311