This is a parts list to ass assemble a recovery chamber to incubate acute brain slices prepared as described in Oriol Pavon Arocas, Tiago Branco (2021). Preparation of acute midbrain slices for patch-clamp recordings. [https://dx.doi.org/10.17504/protocols.io.bukhnut6].
To assemble a recovery chamber you will need the following:
- Glass beaker, 150 mL, 60 mm external diameter (Borosilicate glass with spout, Cat. No. 213-1123, VWR).
- Microfilter candles (or bubblestones) to bubble solutions: 6 mm diameter, porosity P3, narrow glass tube (Cat. No. ROBU18103, Robuglas).
- Plastipak luer slip syringe (2 mL, sterile, Cat. No BD574 or BDS0003, Appleton Woods). We cut the tip so it has the right length and insert a stainless steel needle (e.g. brown - BD Microlance 3, 26G, 0.45x13 mm, Cat. No. 303800) at the bottom to make a support for the bubblestone.
- Sterile gauze swabs (10x10 cm, 8 ply, Cat. No. MSPPEJE136 or IBSA25071, VWR).
- Double threaded plastic cap (Samco™ Narrow Mouth Bio-Tite™ 90mL (3 oz.) 48mm Specimen Containers, Sterile, Cat. No. 020039, Thermo Fisher).
Alternatively, if plastic caps of the correct size can't be found, you can use the 3D printed pieces available at this repository.
- Recovery_Chamber.step - design of the pieces. They fit the 150 mL beaker listed above.
- Recovery_Chamber_outer_piece.stl - we print it on a Ultimaker S5.
- Recovery_Chamber_inner_piece.stl - we print it on a Ultimaker S5.
The 3D printed pieces we currently use for the recovery chamber were designed by Robb Barrett from the Fabrications Lab at the Sainsbury Wellcome Centre (UCL) and can be downloaded from this repository. The .stl and .step files are made available under a CC0-1.0 license. We hope this will be a far cheaper and easy to assemble alternative to the commercially available recovery chambers.
Below are some images of how the 3D printed pieces and the assembled recovery chamber look like:
