Two-Photon Fluorescent Probe for Mitochondrial Zinc Imaging
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Solution Overview
Problem
Current one-photon fluorescent probes lack selectivity for mitochondria and suffer from shallow penetration depth and photo bleaching, limiting their effectiveness in visualizing zinc ions within cells and tissues.
Innovation Solution
Development of a two-photon fluorescent probe (SZn-Mito) that selectively detects zinc ions in mitochondria using a compound structure incorporating triphenylphosphonium salt and N,N-di-(2-picolyl)ethylenediamine, allowing for deeper tissue penetration and prolonged imaging.
Engineering Contradictions & Design Principles
Engineering Contradiction Analysis
1Measurement precision
If one-photon fluorescent probes are used to detect zinc ions, then fluorescence detection can be achieved, but the probes lack selectivity for mitochondria and show poor selectivity to zinc ions
Solution Approach 1:
The patent combines multiple functional components into a single probe molecule: a two-photon fluorescent core (BTDAN or BODAN) for deep tissue imaging, a zinc ion chelating unit (dipicolylamine) for selective zinc detection, and a mitochondria-targeting unit (triphenylphosphonium salt) for mitochondrial localization. This merging of functions into one integrated probe resolves the selectivity issues of previous separate probes
Solution Approach 2:
The probe employs a composite molecular structure combining different functional moieties: the two-photon fluorophore (benzo[d]thiazol-2-yl)-2-(N,N-dimethylamino)naphthalene or (benzo[d]oxazol-2-yl)-2-(N,N-dimethylamino)naphthalene, the zinc-chelating dipicolylamine group, and the mitochondria-targeting triphenylphosphonium salt. This composite structure enables simultaneous achievement of deep penetration, zinc selectivity, and mitochondrial localization
2Length of moving object
If one-photon fluorescent probes are used, then zinc ion detection is possible, but the excitation wavelength is short leading to shallow penetration depth and photo bleaching
Solution Approach 1:
The patent changes the excitation wavelength parameter from the ultraviolet/blue range (300-480 nm) used by one-photon probes to the near-infrared range (700-900 nm) for two-photon excitation. This parameter change enables deeper tissue penetration due to reduced scattering and absorption in the near-infrared window, and reduces photo bleaching and phototoxicity
Solution Approach 2:
The patent replaces the one-photon excitation mechanism with a two-photon excitation mechanism. Instead of using a single high-energy photon, the system uses two lower-energy near-infrared photons simultaneously absorbed by the fluorophore to achieve the same electronic transition. This substitution enables deeper penetration and reduced photodamage
3Duration of action of moving object
If conventional fluorescent probes are used, then zinc ion visualization can be achieved, but the imaging duration is limited due to photo bleaching
Solution Approach 1:
By changing the excitation wavelength to near-infrared for two-photon excitation, the patent reduces the energy per photon and avoids the photodamaging effects associated with high-energy UV/blue light. This parameter change extends the imaging duration by reducing photo bleaching and phototoxicity, enabling prolonged observation of zinc ion dynamics in living cells
Applied Scientific Principles
This section explains which scientific principles are used to turn an abstract innovation direction into a practical engineering solution.
Function Achieved in This Case
SZn-Mito enables selective visualization and prolonged detection of zinc ions within mitochondria, offering improved sensitivity and stability, with a deeper penetration depth and longer imaging duration compared to conventional probes.
Implementation Method 1
two-photon fluorescent probe (SZn-Mito) capable of selectively detecting zinc ions present inside the mitochondria
Implementation Method 2
observing the fluorescence with a two-photon microscope
Data Source
AI summary
Provided is a two-photon fluorescent probe, and more particularly, a two-photon fluorescent probe which is one or more selected from compounds represented by Formulae 1 and 2, a method for manufacturing the same, and an imaging method of zinc ions within the mitochondrion using the same. Since two probes are introduced into one molecule, the two-photon fluorescent probe of the present invention can selectively dye the mitochondria, simultaneously with reacting with zinc ions, thereby generating intense fluorescence. Thus, the two-photon fluorescent probe of the present invention can be used for the imaging of zinc ion distribution and activation within the mitochondrion in living cells or intact biological tissues.