Through‐Bond Energy Transfer Cassette with Dual‐Stokes Shifts for “Double Checked” Cell Imaging

Abstract

Organic dyes generally suffer from small Stokes shift that usually leads to self‐quenching and ‐gaining errors during the fluorescent imaging process. Here, a through‐bond energy transfer (TBET) cassette is developed with large Stokes shift to pursue precise cell imaging. The TBET is constructed by covalently conjugated tetraphenylethene (acts as donor) and rhodamine (acceptor) through an acetylene bond. The constructed TBET cassette distinctly behaves as dual‐Stokes shifts, including a large pseudo‐Stokes shift caused by energy transfer, from donor's absorption to acceptor's emission (up to 260 nm) and a smaller Stokes shift of acceptor molecules itself. Due to the intrinsic dual‐Stokes shifts, TBET cassette exhibits specific “dual distinct absorbances, single shared emission” properties, which can be excitated under two different laser channels. By colocalization of the imaging readouts of these two channels, the precisely “double checked” fluorescent imaging is achieved in living cells.

Regulation of acetylcholinesterase during the lipopolysaccharide‐induced inflammatory responses in microglial cells

“…Moreover, the localization of AChE and α7 nAChR was highly co‐localized in primary microglial cells (Figure 2A). The co‐localization efficiency of AChE and α7 nAChR was quantified by two widely used indexes—Pearson's coefficient (PC) and Mander's overlap coefficient (MOC) 29,30 . The values of PC were ~0.75 in BV2 cell and ~0.4 of in rat microglia; while the values of MOC were over 0.9 for both cell types, suggesting a good co‐localization of AChE and α7 nAChR (Figure 2B).…”

“…The co-localization efficiency of AChE and α7 nAChR was quantified by two widely used indexes-Pearson's coefficient (PC) and Mander's overlap coefficient (MOC). 29,30 The values of PC were ~0.75 in BV2 cell and ~0.4 of in rat microglia; while the values of MOC were over 0.9 for both cell types, suggesting a good co-localization of AChE and α7 nAChR (Figure 2B). In addition, ChAT, the critical enzyme responsible for the synthesis of ACh, was detected in BV2 and microglial cells, and the ChAT-positive cells occupied ~70% of the total cell number in both cell types (Figure 2C,D).…”

Simultaneous dual-colour tracking lipid droplets and lysosomes dynamics using a fluorescent probe

“…Owing to their high selectivity, sensitivity, and ease of use, fluorescent probes have assumed central roles within both the chemical and biological communities 5. Traditionally, probes were designed to show specific response toward a specific analyte or class of analytes.…”

A 2-(benzo[d]thiazol-2-yl)phenol based on conjugated polymer: Highly selective colorimetric fluorescent chemosensor for F-depending on Si–O bond cleavage reaction

“…These results are in agreement with those reported previously which also suggest that the remarkable Stokes shift is due to the formation of intramolecular hydrogen bonds cause conformational change of ESIPT state. 59 –62…”