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Plexiglass Drosophila Hybridization Dish
Drosophila brains can be mounted on a 22x22 mm glass coverslip for efficient parallel processing of large numbers of unique samples via fluorescent in situ hybridization (FISH) or immunohistochemistry (Meissner et al., in preparation; Wu et al., 2016). FlyLight and Janelia Experimental Technology previously described a Drosophila Mounting T-Dish that aids the mounting process (https://www.janelia.org/open-science/drosophila-mounting-t-dish). Most coverslip processing steps are carried out by moving coverslips between jars of solutions, but the 10 mL jar volume is too large for the hybridization step in FISH, which requires a high concentration of probes.
We designed an Inverted Coverslip Hybridization Chamber based on the Drosophila Mounting T-Dish design to address this problem. Samples are mounted on the underside of the coverslip, which is held above the chamber on 0.5 mm spacers to each side, creating a 150 µL space for hybridization solution. The opening at the top allows easy application and removal of the coverslip. Minimal evaporation occurs for hours at hybridization temperatures when placed inside a humidified container. Chambers can be washed and reused indefinitely. Although designed for hybridization, the chambers should be suitable for processing coverslips with a small volume of solution.
Meissner GW, Nern A, Singer RH, Wong AM, Malkesman O, Long X. Mapping Neurotransmitter Identity in the Whole-Mount Drosophila Brain Using Multiplex High-Throughput Fluorescence in situ Hybridization. In preparation.
Wu M, Nern A, Williamson WR, Morimoto MM, Reiser MB, Card GM, Rubin GM. Visual projection neurons in the Drosophila lobula link feature detection to distinct behavioral programs. Elife. 2016 Dec 28;5. pii: e21022. doi: 10.7554/eLife.21022.
The information, including CAD files for reproduction, is available for free for non-profit research via the Flintbox link to the right.
The link connects to Flintbox, where you can register and click through a non-commercial license and download the files.
Please reference 2018-034 when inquiring about the Hybridization Dish.