A Wash-Free Cell Membrane Fluorescent Probe Based on Monomer-Excimer Conversion

doi:10.6023/cjoc202511022

Abstract

Excimers, characterized by their large Stokes shifts, broad emission profiles, and longer emission wavelengths compared to monomers, offer significant advantages for bioimaging. A pyrene-based excimer-forming fluorescent probe (BAPI) for wash-free cell membrane imaging was developed. In dilute dimethyl sulfoxide or aqueous solutions, BAPI exhibits weak monomer emission, which transitions to strong excimer emission as the concentration increases. Notably, in various surfactant-containing aqueous solutions, exclusive excimer emission was observed even at very low concentrations. This indicates that BAPI molecules at low-concentration conditions, undergo a monomer-to-excimer conversion upon migrating from the aqueous phase into micelles. Capitalizing on this property, BAPI serves as an effective membrane-targeting probe with several distinct advantages, including a large Stokes shift (129 nm), high fluorescence efficiency (Φ=75.66% in dimethyl sulfoxide), low cytotoxicity, rapid cellular internalization (≈5 min), and wash-free imaging capability.

Key words: cell membrane, fluorescent probe, excimer, wash-free imaging, pyrene

Cite this article

Bailin Guo, Sainü Wang, Bingxin Lei, Enju Wang. A Wash-Free Cell Membrane Fluorescent Probe Based on Monomer-Excimer Conversion[J]. Chinese Journal of Organic Chemistry, 2026, 46(5): 2082-2089.

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Fig. & Tab. 11

Scheme 1. Synthesis of the cell membrane probe BAPI

Figure 1. Emission spectra of BAPI in DMSO (A) and water (B) at a range of concentrations under 370 nm UV irradiation

Figure 2. Absorption spectra of BAPI in aqueous solution at different concentrations

Figure 3. Emission spectra of BAPI (5 μmol/L) in aqueous solution recorded immediately (0 d) and after 7 d under 370 nm UV irradiation, along with the corresponding fluorescence images

Figure 4. (A) Emission spectra of solid-state BAPI excited at 370 nm and the corresponding photos under 365 nm UV irradiation; (B) Emission spectra of 10 μmol/L BAPI in THF/EtOAc mixtures with increasing volume fractions of EtOAc (fEtOAc) excited at 370 nm: (C) Photos under 365 nm UV irradiation when fEtOAc is 10% and 90%

Figure 5. (A) Emission spectra of 5 μmol/L BAPI in aqueous solutions of SDS (20 mmol/L), CTAB (100 mmol/L), CAPB (50 mmol/L) and F127 (100 mmol/L) under 370 nm UV irradiation; (B) Emission spectra of BAPI at various concentrations in 20 mmol/L SDS solution under 370 nm UV irradiation

Figure 6. Viability of HeLa cells treated with different concentrations of BAPI for 24 h

Figure 7. Confocal images of HeLa cells co-incubated with (A~D) BAPI (3.0 μmol/L, 20 min) and DiI (3.0 μmol/L, 15 min) and (E~H) Hoechst 33342 (20 min) and BAPI (3.0 μmol/L, 20 min) (A) Brightfield, (B) green channel (λex=488 nm), (C) red channel (λex=561 nm), and (D) merge of A~C. (E) brightfield, (F) blue channel (λex=405 nm), (G) green channel (λex=488 nm), and (H) merge of E~G.

Figure 8. Confocal images of HeLa cells treated with BAPI (A, B) or DiI (C, D) Images in panels A and C were taken after washing to remove extracellular dye, whereas images in panels B and D were taken without washing.

Figure 9. Confocal images of HeLa cells treated with 5 μmol/L BAPI for (A) 1, (B) 5, (C) 15 and (D) 30 min

Figure 10. Confocal images of HeLa cells treated with different concentrations of BAPI for 5 min (A) 1 µmol/L, (B) 2 µmol/L, (C) 5 µmol/L and (D) 10 µmol/L

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