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2 protocols using methoxypolyethyleneglycol amine 750

1

Synthesis of Naphthalene-Based Fluorescent Dyes

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N,N-Dimethylethylenediamine
and succinic acid disodium salt
were purchased from Nacalai Tesque (Kyoto, Japan). 1,8-Naphthalic
anhydride, 3-nitro-1,8-naphthalic anhydride, iodomethane, and methoxypolyethylene
glycol amine 750 were purchased from Sigma-Aldrich Japan (Tokyo, Japan).
4-Bromo-1,8-naphthalic anhydride, naphthalene-1,4,5,8-tetracarboxylic
dianhydride, and sodium methoxide were purchased from Tokyo Chemical
Industries (Tokyo, Japan). Copper(II) sulfate pentahydrate and sodium
polyacrylate (DP: 22,000–70,000) were purchased from Wako Pure
Chemical Industries (Osaka, Japan). Other reagents were of commercial
grade and used without further purification.
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2

Carboxylated Polystyrene Nanoparticle Functionalization

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Carboxylated polystyrene NPs of different sizes were purchased from Life technologies (Carlsbad, CA, USA). Specifically, 20 nm (F8783), 40 nm (F8793), 100 nm (F8797) and 500 nm (F8813) NPs with dark red (λexem = 660/680), red (λexem = 580/605), blue (λexem = 350/440) and yellow-green (λexem = 505/515) fluorescence, respectively, were used. Methoxypolyethylene glycol amine 2000 (mPEGamine 2 KDa) (06676), methoxypolyethylene glycol amine 750 (mPEGamine 750 Da) (07966), n-[3-dimethylaminopropyl]-n-ethyl, n-[3-dimethylaminopropyl]-n-ethyl [EDC] (E1769), MES hemisodium buffer (M8902), N-Hydroxysuccinimide (NHS) (56405), and Peroxidase type II from horseradish (P8250-50KU) were purchased from Sigma Aldrich (St. Louis, MO, USA). ImmPACT DAB peroxidase (HRP) substrate (SK-4105) was purchased from Vector laboratories (Burlingame, CA, USA). Slide-A-Lyzer Cassettes (20 K) (66003) were purchased from ThermoFisher scientific (Waltham, MA, USA). Fluorescent mounting media (Vectashield, Vector Labs, Burlingame, Ca, USA)
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