Sodium tetraborate

DPPH, tris (hydroxymethyl) aminomethane (Tris), 2,2′-Azobis (2-amidinopropane) dihydrochloride (AAPH), potassium dihydrogenphosphate (KH₂PO₄), dipotassium hydrogenphosphate (K₂HPO₄), ethanol (99.5%), methanol (HPLC grade), hydrochloric acid, sodium dihydrogenphosphate (NaH₂PO₄), disodium hydrogenphosphate (Na₂HPO₄) and sodium tetraborate were purchased from Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Fluorescein sodium salt was purchased from Sigma-Aldrich Japan (Tokyo, Japan), and 6-hydroxy-2,5,7,8-tetramethyl-chroman-2-carboxylic acid (Trolox) was purchased from Tokyo Kasei Kogyo Co., Ltd. (Tokyo, Japan). Luminol and cytochrome c from horse heart were obtained from Nacalai Tesque (Kyoto, Japan).

Azure B (C.I. 52010), basic fuchsin (C.I. 42510), and sodium tetraborate (borax) were purchased from Wako Pure Chemical Industries, Ltd. The following two solutions were prepared.
(A) Staining dye mixture solution: 0.035% Azure B and 0.015% basic fuchsin in 5% aqueous EtOH. First, 0.05% solutions of Azure B and basic fuchsin dissolved in of 5% EtOH were prepared, with the Azure B and basic fuchsin solutions then mixed at a ratio of 7:3 (v/v), respectively.
(B) Borax solution: 0.1 M sodium tetraborate aqueous solution (w/v). A 0.1 M borax solution was prepared by dissolving 3.81 g of sodium tetraborate (Wako Pure Chemical Industries, Ltd) in DW to final volume of 100 mL.
These staining solutions remained stable without any filtration for at least 5 years at room temperature (usually ~20–25°C) in the dark.
The toluidine blue (C.I. 52040; Wako Pure Chemical Industries, Ltd) solution was also prepared as a 0.05% solution in 5% EtOH. toluidine blue staining was performed according to the same protocol used for our staining method.

Pentobarbital was purchased from Sumitomo Dainippon Pharma Co., Ltd. (Toyo, Japan). TUL, cytochalasin D, methyl p-hydroxybenzoate, polyethylene glycol (PEG) 4000, cetyl alcohol, l-menthol, mineral oil, white wax, sodium tetraborate, propylene glycol, beeswax, and sodium dodecyl sulfate were provided by Wako Pure Chemical Industries, Ltd. (Osaka, Japan). Commercially available 0.5 mg TUL tape (CA-TUL, Hokunalin^® Tapes 0.5 mg) was obtained from Mylan EPD G.K (Tokyo, Japan). Rottlerin and dynasore were purchased from Nacalai Tesque (Kyoto, Japan), and methylcellulose (MC, SM-4) was obtained from Shin-Etsu Chemical Co., Ltd. (Tokyo, Japan). 2-hydroxypropyl-β-cyclodextrin (HPβCD) was supplied by Nihon Shokuhin Kako Co., Ltd. (Tokyo, Japan), and PEG 400 was provided by Maruishi Pharmaceutical Co., Ltd. (Osaka, Japan). Nystatin and 450 nm pore size MF^TM membrane filters were purchased from Sigma-Aldrich Japan (Tokyo, Japan) and Merck Millipore (Tokyo, Japan), respectively. All other chemicals used were of the highest purity commercially available.