Chemiluminescence and Bioluminescence Imaging for Biosensing and Therapy: In Vitro and In Vivo Perspectives

Abstract

Chemiluminescence (CL) and bioluminescence (BL) imaging technologies, which require no external light source so as to avoid the photobleaching, background interference and autoluminescence, have become powerful tools in biochemical analysis and biomedical science with the development of advanced imaging equipment. CL imaging technology has been widely applied to high-throughput detection of a variety of analytes because of its high sensitivity, high efficiency and high signal-to-noise ratio (SNR). Using luciferase and fluorescent proteins as reporters, various BL imaging systems have been developed innovatively for real-time monitoring of diverse molecules in vivo based on the reaction between luciferin and the substrate. Meanwhile, the kinetics of protein interactions even in deep tissues has been studied by BL imaging. In this review, we summarize in vitro and in vivo applications of CL and BL imaging for biosensing and therapy. We first focus on in vitro CL imaging from the view of improving the sensitivity. Then, in vivo CL applications in cells and tissues based on different CL systems are demonstrated. Subsequently, the recent in vitro and in vivo applications of BL imaging are summarized. Finally, we provide the insight into the development trends and future perspectives of CL and BL imaging technologies.

Keywords: bioimaging; bioluminescence; biomedicine; biosensing; chemiluminescence.

Figure 1

CL imaging of cancer cells using multifunctional nanoprobes. (A) Glycan expression-based CL imaging of HCCC-9810 cells using AuNP probes. Reprinted with permission from , Copyright 2012, American Chemical Society. (B) Dual-aptamer recognition-based CL imaging of Ramos cells using bio-bar-code AuNP probes. Reprinted with permission from , Copyright 2013, Royal Society of Chemistry. (C) Folic acid and HRP-bifunctionalized semiconducting polymer dots for both CL imaging and PDT of tumor cells. Reprinted with permission from , Copyright 2014, American Chemical Society.

Figure 2

Tool enzyme-based isothermal amplification for CL imaging of nucleic acids. (A) Automated on-chip CL imaging platform based on RPA for simultaneous detection of viruses and bacteria. Reprinted with permission from , Copyright 2016, American Chemical Society. (B) CL imaging for simultaneous amplified detection of three microRNAs via programmable strand displacement-based magnetic separation. Reprinted with permission from , Copyright 2017, Elsevier B.V.

Figure 3

The structure of 3D-printed unibody immunoarray for detection of PSA and PF-4. Reprinted with permission from , Copyright 2017, Royal Society of Chemistry.

Figure 4

Polymer NPs-based CL imaging of in vivo H₂O₂. (A) BLSA/CPPO NPs with aggregation-enhanced fluorescence for CL imaging of H₂O₂. Reprinted with permission from , Copyright 2012, American Chemical Society. (B) Synthesis and application of HPOX micelles in H₂O₂ imaging. Reprinted with permission from , Copyright 2012, Wiley-VCH.

Figure 5

POCL polymer NPs-based CRET systems for in vivo H₂O₂ imaging. (A) Molecular components of CF-polymer NPs and the mechanism of ONOO^-/^-OCl and H₂O₂ detection by CF-polymer NPs. Reprinted with permission from , Copyright 2014, Nature Publishing Group. (B) Mechanism of the CRET in semiconducting POCL polymer NPs and the comparison between PFPV-based semiconducting POCL polymer NPs and other four semiconducting-based POCL polymer NPs, as well as in vivo CL imaging of endogenous H₂O₂ in the mouse model of neuroinflammation. Reprinted with permission from , Copyright 2016, American Chemical Society. (C) O₂^●- sensing and the structure of polymer PCLA-O₂^●-, as well as the in vivo CL imaging of endogenous O₂^●-. Reprinted with permission from , Copyright 2016, American Chemical Society.

Figure 6

Silica NPs-based CL assays for in vivo H₂O₂ imaging. (A) Fabrication of POCL silica nanodevice for H₂O₂ imaging which was generated by different oxidase-catalyzed biomarkers. Reprinted with permission from , Copyright 2017, Wiley-VCH. (B) Synthesis of the HRP-SiO₂@FLuc NPs for in situ sequential imaging of endogenous and exogenous H₂O₂. Reprinted with permission from , Copyright 2016, Wiley-VCH.

Figure 7

1,2-dioxetane based CL probes for CL imaging. (A) In vivo imaging of H1299 lung tumor xenografts using HyCL-2 under different oxygen concentrations. Reprinted with permission from , Copyright 2016, American Chemical Society. (B) CL mechanism of dioxetane-fluorophore conjugates. Reprinted with permission from , Copyright 2016, American Chemical Society.

Figure 8

In situ formation of luciferase and the mating scheme of S1P₁ luciferase signaling mice. Reprinted with permission from , Copyright 2017, Nature Publishing Group.

Figure 9

BL imaging with luciferase mutants. (A) Crystal structure of luciferase with different mutations, the structure of aminoluciferin analogues, and the performance of mutant luciferase with different luciferins in the brains of live mice. Reprinted with permission from , Copyright 2016, Wiley-VCH. (B) Amino acids targeted for mutagenesis and library screening strategy. Reprinted with permission from , Copyright 2017, American Chemical Society.

Figure 10

Luciferin-based conjugates as BL imaging probes. (A) In vivo release of firefly luciferin for endogenous NTR imaging. Reprinted with permission from , Copyright 2016, American Chemical Society. (B) Synthesis of CBP-1 and in vivo imaging of Co²⁺. Reprinted with permission from , Copyright 2018, American Chemical Society. (C) Synthesis of acrylic ester luciferin and in vivo imaging of Cys. Reprinted with permission from , Copyright 2018, American Chemical Society. (D) Synchronous imaging of H₂O₂ and caspase 8 through in situ formation of luciferin. Reprinted with permission from , Copyright 2013, American Chemical Society.

Figure 11

Luciferin analogs for BL imaging. (A) Structures and imaging sensitivity of CycLuc1 compared to _D-luciferin. Reprinted with permission from , Copyright 2014, Nature Publishing Group. (B) Structures, BL emission spectra and tissue penetration efficiency of AkaLumine-HCl, CycLuc1 and _D-luciferin. n = 3, *P < 0.05 (t-test). Reprinted with permission from , Copyright 2016, Nature Publishing Group. (C) Long-term BL imaging of FAAH using (_D-Cys-Lys-CBT)₂. Reprinted with permission from , Copyright 2016, American Chemical Society.

Figure 12

Chemical structures and BL emission of the synthetic luciferins (DTZ and STZ) as well as their corresponding BL imaging in HEK 293T cells and live mice in the presence of various re-engineered luciferases. Reprinted with permission from , Copyright 2017, Nature Publishing Group.