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. Author manuscript; available in PMC: 2013 Oct 9.
Published in final edited form as: Methods Enzymol. 2005;393:288–301. doi: 10.1016/S0076-6879(05)93012-7

TABLE II.

Contents for the Recording Medium (Final Volume of 1 Liter)a

  1. DMEM powderb (13000-021, high glucose, with L-glutamine, with pyrldoxine hydrochloride, without phenol red, without sodium pyruvate, without sodium bicarbonate, Gibco)

  2. Sodium bicarbonate solution (S8761, 7.5%, Sigma) 4.7 ml or 0.35 g culture grade sodium bicarbonate powder

  3. 1 M HEPES buffer (#H0887, Sigma) 10 ml

  4. Penicillin–streptomycin (#15140-122, 10,000 unit/ml–10,000 μg/ml, Gibco) 2.5 ml

  5. B27 supplement (#17504-044, Gibco) 20 ml/or fetal bovine serum 50 ml

  6. Beetle luciferin potassium salt (0.1 mM final concentration, E1602, Promega, stock solution should be made with 0.1 M concentration with sterilized water and a small aliquot should be kept −80° with light protection)

a

All contents (except Nos. 5 and 6) should be dissolved in autoclaved Milli-Q water, and total volume should be adjusted to 1 liter. pH should be stable at around 7 in a few days and osmolality should be around 300. Medium should be kept light protected at 4°. Luciferin should be added, and necessary amounts of the medium should be warmed up to 37°.

b

We used this medium in all experiments published in 2000–2004. However, DMEM powder has been discontinued and is only available through custom orders. The DMEM powder (#13000-021 Gibco) can be replaced with DMEM power (#D-2902, with L-glutamine and 1000 mg glucose, without phenol red and sodium bicarbonate, Sigma) and 3.5 g of D-glucose powder (G7021, Sigma).