Bioluminescence imaging: progress and applications

Badr, Christian E.; Tannous, Bakhos A.

doi:10.1016/j.tibtech.2011.06.010

Cited by 253 publications

(216 citation statements)

References 85 publications

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“…In this instantiation, both the intensity and spectral properties of bioluminescent proteins are altered when they are associated with fluorescent proteins in a process termed bioluminescence resonance energy transfer. [31][32][33] Luciferase proteins have also [34][35][36][37][38] Given the expanding toolbox of bioluminescent proteins and their wide variety of applications (see reviews by Badr and Tannous 39 and Saito and Nagai 40 ), it is timely to look at methods of detection of bioluminescence in laboratories not necessarily set up for bioluminescence imaging per se or unwilling to purchase commercially available bioluminescence imagers (e.g., Olympus LV200 microscope, IVIS Spectrum animal imager). In this paper, we discuss advantages and disadvantages of the various methods we have utilized for detection and quantification of bioluminescent signals from live cells and animals.…”

Section: Introductionmentioning

confidence: 99%

Bioluminescence imaging in live cells and animals

et al. 2016

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Abstract. The use of bioluminescent reporters in neuroscience research continues to grow at a rapid pace as their applications and unique advantages over conventional fluorescent reporters become more appreciated. Here, we describe practical methods and principles for detecting and imaging bioluminescence from live cells and animals. We systematically tested various components of our conventional fluorescence microscope to optimize it for long-term bioluminescence imaging. High-resolution bioluminescence images from live neurons were obtained with our microscope setup, which could be continuously captured for several hours with no signs of phototoxicity. Bioluminescence from the mouse brain was also imaged noninvasively through the intact skull with a conventional luminescence imager. These methods demonstrate how bioluminescence can be routinely detected and measured from live cells and animals in a cost-effective way with common reagents and equipment. © The Authors. Published by SPIE under a Creative Commons Attribution 3.0 Unported License. Distribution or reproduction of this work in whole or in part requires full attribution of the original publication, including its DOI.

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Section: Introductionmentioning

confidence: 99%

Bioluminescence imaging in live cells and animals

et al. 2016

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“…Four hours later, the animals were sacrificed by CO 2 , and target organs were removed and frozen on dry ice. For the luciferase assay, the tissues were homogenized in 1 mL of 5× cell culture lysis buffer (Promega), incubated on ice for 5 min, and then spun in a centrifuge for 5 min.…”

Section: Tissue Homogenates and Luciferase Assaymentioning

confidence: 99%

“…Because mammalian tissues do not naturally emit bioluminescence, in vivo BLI has considerable appeal because images can be generated with very little background signal, and BLI is emerging as a powerful technology for multiple therapeutic areas. [1][2][3] The utility of reporter gene technology makes it possible to analyze specific cellular and biological processes in a living animal through imaging methods. 1 For several years, bioluminescence-based reporter gene assays have been employed to measure functional activity of G-proteincoupled receptors (GPCRs).…”

Section: Introductionmentioning

confidence: 99%

The CRE luc Bioluminescence Transgenic Mouse Model for Detecting Ligand Activation of GPCRs

et al. 2014

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Numerous assays have been developed to investigate the interactions between G-protein-coupled receptors (GPCRs) and their ligands since GPCRs are key therapeutic targets. Reporter-based assays using the cAMP response element (CRE) coupled with bioluminescence from a luciferase reporter have been used extensively in vitro with high-throughput screens (HTS) of large chemical compound libraries. We have generated a transgenic mouse model (CRE luc) with a luciferase reporter under the control of a synthetic promoter that contains several CREs, which supports real-time bioimaging of GPCR ligand activity in whole animals, tissues, or primary cells. In the CRE luc model, GPCR signaling through the cAMP pathway can be detected from the target GPCR that is in a native cellular environment with a full complement of associated receptors and membrane constituents. Multiple independent lines have been produced by random integration of the transgene, resulting in tissue expression profiles covering the major organs. The goal of the CRE luc model is to accelerate the transition from HTS to profiling of GPCR small-molecule leads in preclinical animal disease models, as well as define the mechanism of action of GPCR drugs in three experimental formats: primary cells, tissue homogenates, and whole animals.

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“…One set of noninvasive imaging methods take advantage of genetically introduced imaging reporters in cancer cells to track their location in vivo. These techniques include fluorescence, bioluminescence, positron emission tomography (PET), and single-photon emission computed tomography (SPECT) [8][9][10] . †Electronic supplementary information (ESI) available.…”

Section: Introductionmentioning

confidence: 99%

Quantifying spontaneous metastasis in a syngeneic mouse melanoma model using real time PCR

Deng

McLaughlin

Klinke

2017

Preprint

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Modeling metastasis in vivo with animals is a priority for both revealing mechanisms of tumor dissemination and developing therapeutic methods. While conventional intravenous injection of tumor cells provides an efficient and consistent system for studying tumor cell extravasation and colonization, studying spontaneous metastasis derived from orthotopic tumor sites has the advantage of modeling more aspects of the metastatic cascade, but is challenging as it is difficult to detect small numbers of metastatic cells. In this work, we developed an approach for quantifying spontaneous metastasis in the syngeneic mouse B16 system using real time PCR. We first transduced B16 cells with lentivirus expressing firefly luciferase Luc2 gene for bioluminescence imaging. Next, we developed a real time quantitative PCR (qPCR) method for the detection of luciferase-expressing, metastatic tumor cells in mouse lungs and other organs. To illustrate the approach, we quantified lung metastasis in both spontaneous and experimental scenarios using B16F0 and B16F10 cells in C57BL/6Ncrl and NOD-Scid Gamma (NSG) mice. We tracked B16 melanoma metastasis with both bioluminescence imaging and qPCR, which were found to be self-consistent. Using this assay, we can quantitatively detect one Luc2 positive tumor cells out of 10 4 tissue cells, which corresponds to a metastatic burden of 1.8x10 4 metastatic cells per whole mouse lung. More importantly, the qPCR method was at least a factor of 10 more sensitive in detecting metastatic cell dissemination and should be combined with bioluminescence imaging as a high-resolution, end-point method for final metastatic cell quantitation. Given the rapid growth of primary tumors in many mouse models, assays with improved sensitivity can provide better insight into biological mechanisms that underpin tumor metastasis.

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Bioluminescence imaging: progress and applications

Cited by 253 publications

References 85 publications

Bioluminescence imaging in live cells and animals

Bioluminescence imaging in live cells and animals

The CRE luc Bioluminescence Transgenic Mouse Model for Detecting Ligand Activation of GPCRs

Quantifying spontaneous metastasis in a syngeneic mouse melanoma model using real time PCR

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