Coumarin 6

Coumarin 6, a fluorescent dye, is used as a fluorescent probe in a microparticle drug delivery system to conduct in vivo tracking, cell uptake, and transport mechanism studies of drug delivery systems (λ_exc=450 nm, λ_em=505 nm)^[1][2][3]. In Vitro: Coumarin 6 labeled HPMC nanoparticles^[3]
(1) Dissolve Coumarin 6 in DMF to prepare a 5 mg/mL organic phase solution.
(2) Dissolve hydroxypropyl methyl cellulose (HPMC) in HBSS to prepare a 50 μg/mL aqueous solution.
(3) Add the organic solution to the aqueous solution using a MasterFlex L/S pump (volume ratio of organic solution to aqueous solution is 1:100) with an injection rate of 36 mL/min.
(4) Prepare 70 nm nanoparticles by stirring at room temperature for 5 min at a stirring speed of 9 mL/min.
(5) Add 2 μg/mL nanoparticles to the MDCKII cell culture dish and incubate in the dark for 5 min, 15 min, 30 min and 60 min.
(6) Add cold PBS buffer to stop the uptake of nanoparticles by MDCKII cells and clean with HBSS.
(7) Use a fluorescence microscope to observe the uptake of nanoparticles by MDCKII cells.