Boosting Bioluminescence Neuroimaging: An Optimized Protocol for Brain Studies

Aswendt, Markus; Adamczak, Joanna; Couillard-Despres, Sebastien; Hoehn, Mathias
February 2013
PLoS ONE;Feb2013, Vol. 8 Issue 2, p1
Academic Journal
Bioluminescence imaging is widely used for optical cell tracking approaches. However, reliable and quantitative bioluminescence of transplanted cells in the brain is highly challenging. In this study we established a new bioluminescence imaging protocol dedicated for neuroimaging, which increases sensitivity especially for noninvasive tracking of brain cell grafts. Different D-Luciferin concentrations (15, 150, 300 and 750 mg/kg), injection routes (iv, ip, sc), types of anesthesia (Isoflurane, Ketamine/Xylazine, Pentobarbital) and timing of injection were compared using DCX-Luc transgenic mice for brain specific bioluminescence. Luciferase kinetics was quantitatively evaluated for maximal photon emission, total photon emission and time-to-peak. Photon emission followed a D-Luciferin dose-dependent relation without saturation, but with delay in time-to-peak increasing for increasing concentrations. The comparison of intravenous, subcutaneous and intraperitoneal substrate injection reflects expected pharmacokinetics with fastest and highest photon emission for intravenous administration. Ketamine/Xylazine and Pentobarbital anesthesia showed no significant beneficial effect on maximal photon emission. However, a strong difference in outcome was observed by injecting the substrate pre Isoflurane anesthesia. This protocol optimization for brain specific bioluminescence imaging comprises injection of 300 mg/kg D-Luciferin pre Isoflurane anesthesia as an efficient and stable method with a signal gain of approx. 200% (compared to 150 mg/kg post Isoflurane). Gain in sensitivity by the novel imaging protocol was quantitatively assessed by signal-to-noise calculations of luciferase-expressing neural stem cells grafted into mouse brains (transplantation of 3,000–300,000 cells). The optimized imaging protocol lowered the detection limit from 6,000 to 3,000 cells by a gain in signal-to-noise ratio.


Related Articles

  • In vivo characterisation of the inflammatory reaction following mesh implantation in transgenic mice models. Fet, N.; Alizai, P.; Fragoulis, A.; Wruck, C.; Pufe, T.; Tolba, R.; Neumann, U.; Klinge, U. // Langenbeck's Archives of Surgery;Jun2014, Vol. 399 Issue 5, p579 

    Introduction: Hernia repair with prosthetic meshes represents one of the most common surgical procedures in the field of surgery. This intervention is always associated with an ensuing inflammatory response, angiogenesis and fibrotic encapsulation forming a foreign body granuloma (FBG) around...

  • Luciferase expression is driven by the promoter of fibroblast activation protein-α in murine pulmonary fibrosis. Wenlong, Li; Leilei, Yang; Wei, Fan; Yi, Chen; Jing, Tian; Lanzhi, Ma; Bing, Liu; Yang, Li; Shaoxia, Wang; Qiuxia, Fu; Zeliang, Chen; Zengming, Zhao // Biotechnology Letters;Sep2015, Vol. 37 Issue 9, p1757 

    Objectives: We used optical imaging of live animals and transgenic technology to develop a pulmonary fibrosis model in mice that can non-invasively and in real-time trace the pulmonary fibrosis process. Results: Fibroblast activation protein-α (FAPα) is selectively expressed in fibrotic...

  • Split-luciferase complementary assay: applications, recent developments, and future perspectives. Azad, Taha; Tashakor, Amin; Hosseinkhani, Saman // Analytical & Bioanalytical Chemistry;Sep2014, Vol. 406 Issue 23, p5541 

    Bioluminescent systems are considered as potent reporter systems for bioanalysis since they have specific characteristics, such as relatively high quantum yields and photon emission over a wide range of colors from green to red. Biochemical events are mostly accomplished through large protein...

  • Seeing Genes at Work in the Living Brain with Non-Invasive Molecular Imaging. Deroose, Christophe M.; Reumers, Veerle; Debyser, Zeger; Baekelandt, Veerle // Current Gene Therapy;Jun2009, Vol. 9 Issue 3, p212 

    Over the past ten years, a variety of imaging techniques have been developed that allow non-invasive detection of gene expression within the brain of intact mammals, ranging from mouse to man. The basic concepts of these imaging techniques, including positron emission tomography, single photon...

  • Role of miRNA-146 in proliferation and differentiation of mouse neural stem cells. Wei-Zhong Xiao; An-Qing Lu; Xin-Wei Liu; Zhe Li; Ying Zi; Zhi-Wei Wang // Bioscience Reports;Oct2015, Vol. 35 Issue 5, p1 

    Neural stem cells (NSCs) have been defined as neural cells with the potential to self-renew and eventually generate all cell types of the nervous system. NSCs serve as an ideal cell type for nervous system repair. In the present study, miR-146 overexpression and predicted target (notch 1) were...

  • Neural stem cell-mediated CE/CPT-11 enzyme/prodrug therapy in transgenic mouse model of intracerebellar medulloblastoma. Gutova, M; Shackleford, G M; Khankaldyyan, V; Herrmann, K A; Shi, X-H; Mittelholtz, K; Abramyants, Y; Blanchard, M S; Kim, S U; Annala, A J; Najbauer, J; Synold, T W; D'Apuzzo, M; Barish, M E; Moats, R A; Aboody, K S // Gene Therapy;Feb2013, Vol. 20 Issue 2, p143 

    Medulloblastoma is a heterogeneous diffuse neoplasm that can be highly disseminated, and is the most common malignant childhood brain tumor. Although multimodal treatments have improved survival rates for patients with medulloblastoma, these tumors are associated with high morbidity and...

  • Visualization of spatiotemporal differentiation of dopaminergic interneurons in adult mouse olfactory bulb using transgenic mice. Saino-Saito, Sachiko // Anatomical Science International;Dec2008, Vol. 83 Issue 4, p228 

    Olfactory bulb (OB) interneurons, predominantly periglomerular (PG) and granule (GR) cells, are derived from neural stem cells in the subventricular zone (SVZ) throughout life and migrate to the OB in the rostral migratory stream (RMS). In the adult superficial GR layer transgene expression was...

  • "Color Timer" mice: visualization of neuronal differentiation with fluorescent proteins. Kanki, Hiroaki; Shimabukuro, Marilia Kimie; Miyawaki, Atsushi; Okano, Hideyuki // Molecular Brain;2010, Vol. 3, p1 

    The molecular mechanisms governing the differentiation of neural stem cells (NSCs) into neuronal progenitor cells and finally into neurons are gradually being revealed. The lack of convenient means for real-time determination of the stages of differentiation of individual neural cells, however,...

  • In Vivo Fate Analysis Reveals the Multipotent and Self-Renewal Features of Embryonic AspM Expressing Cells. Marinaro, Cinzia; Butti, Erica; Bergamaschi, Andrea; Papale, Alessandro; Furlan, Roberto; Comi, Giancarlo; Martino, Gianvito; Muzio, Luca // PLoS ONE;2011, Vol. 6 Issue 4, p1 

    Radial Glia (RG) cells constitute the major population of neural progenitors of the mouse developing brain. These cells are located in the ventricular zone (VZ) of the cerebral cortex and during neurogenesis they support the generation of cortical neurons. Later on, during brain maturation, RG...


Read the Article


Sorry, but this item is not currently available from your library.

Try another library?
Sign out of this library

Other Topics