Endoplasmic Reticulum-Directed Ratiometric Fluorescent Probe for Quantitive Detection of Basal H₂O₂

Abstract

The endoplasmic reticulum (ER) has a central role in the fine-tuning of environmental and internal stimuli. We herein report a ratiometric fluorescent probe, α-Naph, capable of determining basal HO in the ER. The probe specifically responds to HO. The limit of detection of the probe is as low as 38 nM, making it a feasible sensor to image intracellular basal HO. In addition, utilizing its ratiometric property, we are able to measure the concentration of HO in the ER quantitatively, eliminating the error caused by the probe concentration and environment. The intracellular concentration of HO in the ER is calculated to be 0.692 μM under normal conditions and 1.26 μM under the stimulation of phorbol myristate acetate.

A guide to assessing endoplasmic reticulum homeostasis and stress in mammalian systems

“…To overcome this caveat, several modifications of disulfide‐based reporters and protocols were suggested . A better way to solve this issue would be the development of disulfide independent ER probes that are actively being searched for . Peroxide probes will however never report on H 2 O 2 levels as dynamically and accurately as GSH/GSSG reporting probes.…”

Red AIE‐Active Fluorescent Probes with Tunable Organelle‐Specific Targeting

“…All the spectroscopic grade solvents such as acetonitrile (ACN), ethanol (EtOH), tetrahydrofuran (THF), dichloromethane (DCM), ethyl acetate (EA), hexane, and DMSO were procured from the Merck Company. All the molecules synthesized were purified by column chromatography and recrystallized using double layer solution diffusion with dichloromethane/hexane or dichloromethane/ethyl acetate and fully characterized by 1 H NMR, 13 C NMR, and HRMS. 1 H and 13 C NMR spectra were recorded on a Bruker AV 400 Spectrometer at 400 and 100 MHz in CDCl 3 and d 6 -DMSO, respectively.…”

“…[51] Finally, the targeted fluorophores CDPP-3SO 3 , CDPP-4SO 3 , and CDPP-BzBr, were synthesized by simple reactions between CDPP and 1,3-propane sultone, 1,4-butane sultone, and 4-bromobenzyl bromide, respectively (Figure 2A; Scheme S1, Supporting Information). [51][52][53] The chemical structures of all the synthesized compounds were characterized by standard spectroscopic techniques such as 1 H NMR, 13 C NMR, and high-resolution mass spectroscopy (HRMS) (Figures S1-S5, Supporting Information).…”

“…With the development of fluorescent bioprobes, researchers became able to visualize the once invisible organelles revealing important and significant insight . Development of the bioprobes has resulted in organelle specific staining agents allowing researchers to study the targeted organelle . The visualization techniques reveal the location and morphology of the organelles, and thus enable the study and monitoring of changes that reveal important biological states or dysfunction .…”

“…[9,10] Development of the bioprobes has resulted in organelle specific staining agents allowing researchers to study the targeted organelle. [11][12][13][14][15] The visualization techniques reveal the location and morphology of the organelles, and thus enable the study and monitoring of changes that reveal important biological states or dysfunction. [16] In particular, super-resolution microscopy is able to provide unprecedented clarity into the morphology of these structures.…”