Two-photon fluorescence lysosomal bioimaging with a micelle-encapsulated fluorescent probe

Area of Science:

• Biomedical Optics
• Materials Science
• Cell Biology

Background:

• Lysosomes are crucial organelles involved in cellular degradation.
• Developing selective fluorescent probes is essential for studying lysosome function.
• Two-photon microscopy offers advantages for deep tissue imaging and reduced phototoxicity.

Purpose of the Study:

• To synthesize and characterize a novel two-photon absorbing (2PA) fluorescent probe for selective lysosome imaging.
• To evaluate the probe's efficiency and performance within a micellar delivery system.
• To assess the probe's utility in conjunction with two-photon fluorescence microscopy (2PFM).

Main Methods:

• Microwave-assisted synthesis was employed for efficient preparation of the 2PA fluorescent probe (dye I).
• Dye I was encapsulated within Pluronic® F-127 micelles for enhanced delivery and stability.
• Photophysical characterization was performed to determine the probe's optical properties, including 2PA efficiency.
• In vitro cell viability assays were conducted to ensure biocompatibility.
• Two-photon fluorescence microscopy (2PFM) was utilized for imaging.

Main Results:

• The microwave-assisted synthesis yielded improved efficiency and reduced reaction times for dye I.
• The synthesized probe exhibited significant two-photon absorption efficiency.
• The Pluronic® F-127 micelle formulation provided a suitable delivery system for the probe.
• Preliminary imaging and cell viability studies indicated successful lysosome targeting and good biocompatibility.

Conclusions:

• A novel, efficient, lysosome-selective two-photon absorbing fluorescent probe has been successfully developed.
• The probe, delivered via Pluronic® F-127 micelles, is suitable for two-photon fluorescence microscopy.
• This system holds potential for advanced cellular imaging and investigation of lysosomal functions.