BRET based dual-colour (visible/near-infrared) molecular imaging using a quantum dot/EGFP–luciferase conjugate

Abstract

A bioluminescent dual-colour molecular-imaging probe was prepared to emit green and near-infrared luminescence from a conjugate between enhanced green fluorescent protein (EGFP), Renilla luciferase (RLuc) and CdSeTe/CdS quantum dot (QD).

Shortwave-infrared (SWIR) fluorescence molecular imaging using indocyanine green–antibody conjugates for the optical diagnostics of cancerous tumours

“…For tumour imaging, we prepared two types of cancer model mice, where skin cancer cell (A431) 30 and breast cancer cell (KPL-4) 31 were implanted. A431 and KPL-4 cells are the cancer cells established from a human epidermoid carcinoma and a human breast tumour, respectively.…”

“…A431 and KPL-4 cells are the cancer cells established from a human epidermoid carcinoma and a human breast tumour, respectively. 30,31 To check the expression level of the membrane proteins, HER2, EGFR, VEGFR-2, and PD-L1, we rst conducted western blotting analysis for A431 and KPL-4 cells (Fig. 2a).…”

Near-infrared fluorescent protein and bioluminescence-based probes for high-resolution in vivo optical imaging

“…5d-g). 115 The major advantage in the use of QDs as BRET acceptors is the bright emission from the QDs because of the high extinction coefficients and large Stokes shifts. Although most QDs contain heavy metals with cytotoxicity, the improvement of the biocompatibility of QDs is necessary to develop the biological applications of QD-based BRET probes for NIR in vivo imaging.…”

BRED: bioluminescence energy transfer to dye for monitoring ceramide trafficking in cell

“…Bioluminescence resonance energy transfer (BRET) [2][3][4][5] and fluorescence resonance energy transfer (FRET) [6][7][8] have been frequently used for this purpose. However, despite numerous examples, these techniques are limited by low brightness and different maturation speed of RET donors and acceptors [9], poor spectral overlap [10], and a low dynamic range of resonance energy transfer (RET) from a donor to an acceptor within a distance of 2−10 nm [11].…”

“…Another way to increase BRET donor expression may be realized by the use of strong promoters such as human cytomegalovirus (CMV) promoter [23]. Up to now, most of the characterized BRET systems have been established on episomal plasmids [2,3]. A consequence and a major caveat to available BRET systems is the clonal variation of plasmidbased systems, causing a high degree of phenotypic heterogeneity that limits achieving high-level expression, prediction of values, and process streamlining [24].…”