Recent development of two-photon fluorescent probes for bioimaging

Area of Science:

• Biochemistry
• Biophysics
• Chemical Biology

Background:

• Fluorescent probes are vital for biological system research.
• Two-photon excitable probes have gained significant interest for bioimaging due to enhanced tissue penetration and reduced photobleaching compared to one-photon probes.
• Their application in bioimaging is rapidly expanding.

Purpose of the Study:

• To review various types of two-photon excitable probes used in bioimaging.
• To categorize these probes based on design principles and target analytes.
• To provide a foundation for developing novel two-photon probes for critical biological questions.

Main Methods:

• Literature review of two-photon probe development and application.
• Categorization of probes by excitation mechanism (two-photon vs. one-photon).
• Classification based on targeted biological molecules or cellular structures.

Main Results:

• Summary of diverse two-photon probe architectures and their functionalities.
• Analysis of probe performance in various bioimaging contexts.
• Identification of key design strategies enabling two-photon excitation.

Conclusions:

• Two-photon probes offer superior performance for deep-tissue bioimaging.
• Understanding probe design principles is crucial for optimizing imaging capabilities.
• This review facilitates the creation of advanced probes for future biological discoveries.