FITC -Chitosan (MW 100000) is a chitosan (Chitosan) labeled with FITC. FITC -Chitosan (MW 100000) combines the biological and physicochemical properties of chitosan (such as biocompatibility, positive charge, and nanoparticle-forming ability) with the fluorescent visibility of FITC (Ex/Em = ~485/535 nm) [1] [2] [3].
In Vitro, FITC-Chitosan (high molecular weight, e.g., 300 kDa, 18 kDa) can be used to prepare fluorescence-labeled nanoparticles and investigate cellular uptake and in vivo biodistribution of nanoparticles[1][3]. Guide (the following is a recommended protocol for reference only, which can be adjusted according to actual requirements). 1. Cell culture: Seed cells in multi-well plates or confocal culture dishes, and culture them in medium containing 10% FBS until reaching 70-90% confluency. 2. Experimental procedures: 2.1 Wash cells with PBS or serum-free medium. Add fresh serum-free medium containing FITC-Chitosan or its nanoparticles at different concentrations. Key controls: Set up two temperature groups, 37°C (allowing internalization) and 4°C (inhibiting internalization, mainly investigating surface adsorption). 2.2 Incubation: Incubate cells in a cell incubator for 0.5 to 4 hours (determine the linear uptake phase according to pre-experiments). 2.3 Termination and washing: Immediately after incubation, wash cells 3-4 times with pre-cooled PBS to terminate the reaction and remove unbound nanoparticles. 2.4 Membrane surface fluorescence quenching (optional but recommended): To distinguish FITC-Chitosan bound to the cell surface from internalized FITC-Chitosan, digest cells with 0.25% trypsin at 37°C for 30 minutes to remove surface-bound probes, followed by another wash with PBS. 3. Cell lysis and fluorescence quantification: 3.1 Add 0.2 N NaOH solution containing 0.5% Triton X-100 to lyse cells. 3.2 Transfer the cell lysate to a black opaque 96-well plate. Measure the fluorescence intensity using a fluorescence microplate reader at Ex/Em = ~485/535 nm. 3.3 Convert the fluorescence value to the mass of FITC-Chitosan taken up by cells via a FITC-Chitosan standard curve (prepared in the same lysate).
References:
[1]. Huang M, et al. Uptake of FITC-chitosan nanoparticles by A549 cells. Pharm Res. 2002;19(10):1488-1494.
[2]. Cheng X, et al. DNA/chitosan nanocomplex as a novel drug carrier for doxorubicin. Drug Deliv. 2009;16(3):135-144.
[3]. Yan L, et al. Small interfering RNA-loaded chitosan hydrochloride/carboxymethyl chitosan nanoparticles for ultrasound-triggered release to hamper colorectal cancer growth in vitro. Int J Biol Macromol. 2020;162:1303-1310.
In Vitro, FITC-Chitosan (high molecular weight, e.g., 300 kDa, 18 kDa) can be used to prepare fluorescence-labeled nanoparticles and investigate cellular uptake and in vivo biodistribution of nanoparticles[1][3]. Guide (the following is a recommended protocol for reference only, which can be adjusted according to actual requirements). 1. Cell culture: Seed cells in multi-well plates or confocal culture dishes, and culture them in medium containing 10% FBS until reaching 70-90% confluency. 2. Experimental procedures: 2.1 Wash cells with PBS or serum-free medium. Add fresh serum-free medium containing FITC-Chitosan or its nanoparticles at different concentrations. Key controls: Set up two temperature groups, 37°C (allowing internalization) and 4°C (inhibiting internalization, mainly investigating surface adsorption). 2.2 Incubation: Incubate cells in a cell incubator for 0.5 to 4 hours (determine the linear uptake phase according to pre-experiments). 2.3 Termination and washing: Immediately after incubation, wash cells 3-4 times with pre-cooled PBS to terminate the reaction and remove unbound nanoparticles. 2.4 Membrane surface fluorescence quenching (optional but recommended): To distinguish FITC-Chitosan bound to the cell surface from internalized FITC-Chitosan, digest cells with 0.25% trypsin at 37°C for 30 minutes to remove surface-bound probes, followed by another wash with PBS. 3. Cell lysis and fluorescence quantification: 3.1 Add 0.2 N NaOH solution containing 0.5% Triton X-100 to lyse cells. 3.2 Transfer the cell lysate to a black opaque 96-well plate. Measure the fluorescence intensity using a fluorescence microplate reader at Ex/Em = ~485/535 nm. 3.3 Convert the fluorescence value to the mass of FITC-Chitosan taken up by cells via a FITC-Chitosan standard curve (prepared in the same lysate).
References:
[1]. Huang M, et al. Uptake of FITC-chitosan nanoparticles by A549 cells. Pharm Res. 2002;19(10):1488-1494.
[2]. Cheng X, et al. DNA/chitosan nanocomplex as a novel drug carrier for doxorubicin. Drug Deliv. 2009;16(3):135-144.
[3]. Yan L, et al. Small interfering RNA-loaded chitosan hydrochloride/carboxymethyl chitosan nanoparticles for ultrasound-triggered release to hamper colorectal cancer growth in vitro. Int J Biol Macromol. 2020;162:1303-1310.