Electrochemiluminescence (ECL) microscopy is an emerging technique with new applications such as imaging of single entities and cells. Herein, we have developed a bimodal and bicolor approach to record both positive ECL (PECL: light-emitting object on dark background) and shadow label-free ECL (SECL: nonemissive object shadowing the background luminescence) images of single cells. This bimodal approach is the result of the simultaneous emissions of [Ru(bpy)3]2+ used to label the cellular membrane (PECL) and [Ir(sppy)3]3- dissolved in solution (SECL). By spectrally resolving the ECL emission wavelengths, we recorded the images of the same cells in both PECL and SECL modes using the [Ru(bpy)3]2+ (?max = 620 nm) and [Ir(sppy)3]3- (?max = 515 nm) luminescence, respectively. PECL shows the distribution of the [Ru(bpy)3]2+ labels attached to the cellular membrane, whereas SECL reflects the local diffusional hindrance of the ECL reagents by each cell. The high sensitivity and surface-confined features of the reported approach are demonstrated by imaging cell-cell contacts during the mitosis process. Furthermore, the comparison of PECL and SECL images demonstrates the differential diffusion of tri-n-propylamine and [Ir(sppy)3]3- through the permeabilized cell membranes. Consequently, this dual approach enables the imaging of the morphology of the cell adhering on the surface and can significantly contribute to multimodal ECL imaging and bioassays with different luminescent systems.

Bimodal Electrochemiluminescence Microscopy of Single Cells / Knezevic S.; Kerr E.; Goudeau B.; Valenti G.; Paolucci F.; Francis P.S.; Kanoufi F.; Sojic N.. - In: ANALYTICAL CHEMISTRY. - ISSN 1520-6882. - ELETTRONICO. - 95:18(2023), pp. 7372-7378. [10.1021/acs.analchem.3c00869]

Bimodal Electrochemiluminescence Microscopy of Single Cells

Valenti G.;Paolucci F.;
2023

Abstract

Electrochemiluminescence (ECL) microscopy is an emerging technique with new applications such as imaging of single entities and cells. Herein, we have developed a bimodal and bicolor approach to record both positive ECL (PECL: light-emitting object on dark background) and shadow label-free ECL (SECL: nonemissive object shadowing the background luminescence) images of single cells. This bimodal approach is the result of the simultaneous emissions of [Ru(bpy)3]2+ used to label the cellular membrane (PECL) and [Ir(sppy)3]3- dissolved in solution (SECL). By spectrally resolving the ECL emission wavelengths, we recorded the images of the same cells in both PECL and SECL modes using the [Ru(bpy)3]2+ (?max = 620 nm) and [Ir(sppy)3]3- (?max = 515 nm) luminescence, respectively. PECL shows the distribution of the [Ru(bpy)3]2+ labels attached to the cellular membrane, whereas SECL reflects the local diffusional hindrance of the ECL reagents by each cell. The high sensitivity and surface-confined features of the reported approach are demonstrated by imaging cell-cell contacts during the mitosis process. Furthermore, the comparison of PECL and SECL images demonstrates the differential diffusion of tri-n-propylamine and [Ir(sppy)3]3- through the permeabilized cell membranes. Consequently, this dual approach enables the imaging of the morphology of the cell adhering on the surface and can significantly contribute to multimodal ECL imaging and bioassays with different luminescent systems.
2023
Bimodal Electrochemiluminescence Microscopy of Single Cells / Knezevic S.; Kerr E.; Goudeau B.; Valenti G.; Paolucci F.; Francis P.S.; Kanoufi F.; Sojic N.. - In: ANALYTICAL CHEMISTRY. - ISSN 1520-6882. - ELETTRONICO. - 95:18(2023), pp. 7372-7378. [10.1021/acs.analchem.3c00869]
Knezevic S.; Kerr E.; Goudeau B.; Valenti G.; Paolucci F.; Francis P.S.; Kanoufi F.; Sojic N.
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/11585/958941
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