Opening a Gateway for Chemiluminescence Cell Imaging: Distinctive Methodology for Design of Bright Chemiluminescent Dioxetane Probes

Abstract

Chemiluminescence probes are considered to be among the most sensitive diagnostic tools that provide high signal-to-noise ratio for various applications such as DNA detection and immunoassays. We have developed a new molecular methodology to design and foresee light-emission properties of turn-ON chemiluminescence dioxetane probes suitable for use under physiological conditions. The methodology is based on incorporation of a substituent on the benzoate species obtained during the chemiexcitation pathway of Schaap’s adamantylidene–dioxetane probe. The substituent effect was initially evaluated on the fluorescence emission generated by the benzoate species and then on the chemiluminescence of the dioxetane luminophores. A striking substituent effect on the chemiluminescence efficiency of the probes was obtained when acrylate and acrylonitrile electron-withdrawing groups were installed. The chemiluminescence quantum yield of the best probe was more than 3 orders of magnitude higher than that of a standard, commercially available adamantylidene–dioxetane probe. These are the most powerful chemiluminescence dioxetane probes synthesized to date that are suitable for use under aqueous conditions. One of our probes was capable of providing high-quality chemiluminescence cell images based on endogenous activity of β-galactosidase. This is the first demonstration of cell imaging achieved by a non-luciferin small-molecule probe with direct chemiluminescence mode of emission. We anticipate that the strategy presented here will lead to development of efficient chemiluminescence probes for various applications in the field of sensing and imaging.

Ultrasensitive Detection of Salmonella and Listeria monocytogenes by Small‐Molecule Chemiluminescence Probes

“…Positive results are available after 96-144 h(4-6 days), depending on the growth of the bacteria. [29] These new phenoxy-dioxetane luminophores exhibited light emission intensity of up to 3000-fold greater than that of the original Schaapsdioxetanes.Assuch, we sought to utilize our chemiluminescence luminophores to design probes for detection of Salmonella and L. monocytogenes. Rapid alternative methods such as immunoassays or DNAa mplification (PCR in particular) have improved initial time to results but typically still require bacterial pre-enrichment of 16-48 h [10] while the detection time is reduced to ar ange from hours to few minutes.…”

“…Presumptive colonies have to be confirmed after the initial isolation. [25][26][27][28][29][30][31][32][33][34] Ar emarkable enhancement of light emission was obtained by simply improving the emissive nature of the excited species,f ormed during the chemiexcitation of Schaapsdioxetanes. Positive results are available after 96-144 h(4-6 days), depending on the growth of the bacteria.…”

Optical Imaging Approaches to Monitor Static and Dynamic Cell‐on‐Chip Platforms: A Tutorial Review

“…The stable Chemiluminescence of the biological probes is a challenge in this optical imaging approach. The light emission may be imaged using a simple microscope (Figure a) or a compact lensless device …”

Direct Real‐Time Monitoring of Prodrug Activation by Chemiluminescence

“…[7][8][9][10][11][12][13][14] Interest in these unique prodrugs has increased recently due to potential therapeutic utility,imaging enhancement capability,a nd promise for advancing the field of personalized medicine. [23] Thep robes are composed of phenoxy dioxetane luminophores,m asked by at riggering substrate.R emoval of the substrate by an enzyme or an analyte of choice triggers the direct chemiexcitation mechanism of the dioxetane luminophore.T his meant that, for the first-time,a ni ntense lightemission signal could be obtained under physiological conditions to image endogenous molecular activities in vitro and in vivo. [17][18][19][20][21][22] We have recently developed an ew class of chemiluminescent probes with remarkable light-emission efficiency under physiological conditions.…”