Abstract
All the chemicals were of analytical grade and used as received. They are listed in Annex. The phosphate-buffered saline (PBS) (10 mM, pH 7.4) was composed of NaCl 137 mM, KCl 2.7 mM, Na2HPO4 10 mM and KH2PO4 1.8 mM. Washing and blocking buffers of 1% (v/v) BSA were prepared in PBS. The substrate solution for the enzymatic reaction during SECM experiments was made in 0.1 M citrate-phosphate buffer, pH = 5.0, and contained additionally commercial TMB and H2O2 of unknown concentration (not specified by manufacturer).
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Annex
Chemical | Supplier | Formula | Chapter |
|---|---|---|---|
Anti-tyrosinase monoclonal antibody (Ab) T311 | Bio Medical (USA) | 3,4,5 | |
Bovine serum albumin (BSA) | Sigma–Aldrich (Switzerland) | 3,4,5,6 | |
Catechol | Fluka (Switzerland) | C6H4(OH)2 | 3 |
Citric acid | Sigma–Aldrich (Switzerland) | C6H8O7 | 3,4,5 |
Commercial 3,3′,5,5′-tetramethylbenzidine (TMBred) solution | ABRAXIS (USA) | 3,4,5 | |
Cripps Pink apples | local market | 7 | |
Deionized water Milli-Q plus 185 model (produced on site) | Millipore (Switzerland) | H2O | 3,4,5,6,7 |
Dipotassium monohydrogen phosphate | Sigma–Aldrich (Switzerland) | K2HPO4 | 3,4,5,6,7 |
Ethanol | Sigma–Aldrich (Switzerland) | CH3CH2OH | 4,5,6 |
Ferrocenemethanol (FcMeOH) | Fisher Chemicals (Switzerland) | Fe(C5H5)(C5H4)CH2OH | 3,4,5,6,7 |
Formaldehyde solution (4% in PBS) | Alfa Aesar (Karlsruhe, Germany) | H2CO | 3,4,5,6,7 |
Hemoglobin (human) | Sigma–Aldrich (Switzerland) | 6 | |
Hydrogen peroxide (3%) | Sigma–Aldrich (Switzerland) | H2O2 | 3,4,5 |
Hydrogen peroxide, 60% | Fisher Chemicals (Switzerland) | H2O2 | 7 |
L-3,4-dihydroxyphenylalanine (L-DOPA, 99%) | Sigma–Aldrich (Switzerland) | C9H11NO4 | 3 |
Methanol (≥99%) | Merck (Switzerland) | CH3OH | 3,4,5,6,7 |
Monopotassium dihydrogen phosphate | Sigma–Aldrich (Switzerland) | KH2PO4 | 3,4,5,6,7 |
Mouse organ sections | Chang Gung Memorial Hospital (Taiwan) | 5,6 | |
Multiwall carbon nanotubes (MWNTs) (TNSMC1, diameter 6–8 nm; length = 0.5–2 μm; –COOH content = 3.86 wt%) | Chengdu Organic Chemicals Co. Ltd. (Chinese Academy of Sciences, Chengdu, China) | 6 | |
Nitro blue tetrazolium (NBT) | Roche (Switzerland) | C40H30Cl2N10O6 | 4 |
Paraffin-embedded skin tissue micro-arrays (TMA) | US Biomax, Inc. (USA) | 4 | |
Paraffin-embedded thick melanoma and normal skin tissues | Sofia Bio (Bulgaria) | 5 | |
Phosphoric acid | J.T. Baker (USA) | H3PO4 | 3,4,5,6,7 |
Polyethylene glycol (PEG) (Sunbright DSPE-050-PA, molecular weight of PEG chains = 5000) | NOF Cooperation (Japan) | 6 | |
Polytetrafluoroethylene (PTFE) membrane (0.1 μm pore size) | Millipore (Switzerland) | 6 | |
Polyvinylidene difluoride (PVDF) membranes | Bio-Rad (USA) | 3,4,5,6,7 | |
Potassium chloride (≥99.5%) | Fluka (Switzerland) | KCl | 3,4,5,6,7 |
Potassium permanganate | J.T. Baker (USA) | KMnO4 | 6 |
Secondary anti-mouse Abs conjugated with horse radish peroxidase (Abs-HRP) | Abcam (UK) | 3,4,5 | |
Sodium chloride (99.5%) | Fluka (Switzerland) | NaCl | 3,4,5,6,7 |
Sodium hydroxide | J.T. Baker (USA) | NaOH | 6 |
Sodium phosphate monobasic (99%) | Fluka (Switzerland) | NaH2PO4 | 3,4,5,6,7 |
Sulfuric acid | J.T. Baker (USA) | H2SO4 | 6 |
Tris base, 3,3′,5,5′-tetramethylbenzidine (TMB) powder | Sigma–Aldrich (Switzerland) | C16H20N2 | 3,4,5 |
Triton X-100 | Sigma–Aldrich (Switzerland) | 4 | |
Tyrosinase (from mushroom, lyophilized powder, ≥1000 unit/mg solid) | Sigma–Aldrich (Switzerland) | 3,4,5 | |
Ruthenium hexamine chloride | Sigma–Aldrich (Switzerland) | Cl3[Ru(NH3)6] | 5,6 |
Xylene | Sigma–Aldrich (Switzerland) | (CH3)2C6H4 | 4,5,6 |
3,3′-diaminobenzidine (DAB) | Sigma–Aldrich (Switzerland) | C12H14N4 | 4 |
5-bromo-4-chloro-3indolyphosphate (BCIP) | Fisher Chemicals (Switzerland) | C8H6BrClNO4P | 4 |
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Lin, TE. (2019). Experimental Part. In: Soft Probes for Bio-electrochemical Imaging. Springer Theses. Springer, Cham. https://doi.org/10.1007/978-3-030-05758-9_2
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DOI: https://doi.org/10.1007/978-3-030-05758-9_2
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