IN VIVO IMAGING - WHITE PAPERS
Cell Transfection allows researchers to manipulate gene expression in cells. In order to evaluate physiological or pathological processes, your specific protein of interest can be expressed, or depleted by means of cell transfection. Furthermore, the new field of molecular imaging allows visualization of molecular mechanisms of disease in real time, non-invasively with reporter genes. New, flexible methodology makes Cell Transfection affordable and easy. Read more
Multiple luciferases have been cloned and applications in molecular biology and in vivo optical imaging have been identified. This review focuses on features of luciferases and their correspondent substrates called luciferins, commonly used in in vivo molecular imaging. Luciferases and bioluminescence imaging offer a significant advantage... Read more
Molecular imaging of gene function by means of RNA interference (RNAi) is powerful, real time methodology. Targeted RNAi gene knockdown is a highly efficient technique technique in comparison to traditional gene knockout technology, facilitating the study of functional genomics in vitro in cell lines as well as in vivo in live animals. Vector co-expression of RNAi with GFP or firefly luciferase (Luc2) serves as a marker for infectivity and allows tracking and imaging of the RNAi knockdown cells in real time. siRNA lentiviral expression methodology is described. Read more
Lentivirus protocol to make your own luciferase cell lines, Fast and easy.
Labeling cells with a genetically encoded reporter gene (firefly luciferase, red fluorescent protein, transferrin, thymidine kinase, etc.) is the number one choice for in vivo molecular imaging. Various transfection protocols to introduce foreign DNA in cells exist. A virus is nature’s own shuttle of genetic material and therefore viral infection is by far the most efficient transduction method even for resilient cell lines. A lentivirus Read more
An in vivo imaging modalities overview
In vivo imaging in laboratory animals is a relatively new methodology that is gaining fast acceptance in biomedical research. Imaging modalities frequently utilized in in vivo research are mostly adaptations from familiar, clinical imaging instrumentation such as X-ray, ultrasound, CT, SPECT, PET and MRI. However, newer technologies such as Optical Imaging and Intravital Microscopy originated with lab animal applications in mind. The diverse choice in imaging platforms available may foster the question of which modality is the best fit for your research model. Read more
Response to therapy assessment.
Microbubble ultrasound contrast agents have been used for qualitative assessment of solid lesions in a clinical setting for several years. Recent work has described the use of these agents for measurement of the tumor vasculature, primarily as a quantitative endpoint for monitoring patient response to anti-cancer therapy. The size of the microbubble agent (1-8 um) renders them purely intravascular flow tracers, and early work confirmed that the kinetics of these agents in the blood are similar to that of erythrocytes Read more
There is a plethora of choice in reporter tags for non-invasive in vivo optical imaging (fluorescence and bioluminescence) that are currently available from various commercial suppliers. This generates tremendous flexibility on one hand, but dilemma of choice on the other. This manuscript reviews the determining factors and will facilitate decision making using an interactive flow chart. Read more
D-luciferin [(S)-2-(6′-hydroxy-2′-benzothiazolyl)thiazoline-4-carboxylic acid] is the substrate of the North American firefly Photinus Pyralis luciferase and Clickbeetle Red and Clickbeetle Green luciferase. Luciferin is a low molecular weight (318.41 g/mole) organic compound that consists of a benzothiazole moiety attached to a thiazole carboxylic acid moiety. Luciferin is a small molecule which freely diffuses across membranes. When luciferin is injected in vivo Read more
Mouse models for in vivo imaging of cancer established by in vivo grafting of syngeneic or xenogeneic cells in recipient animals are commonly practiced. Success of these studies relies upon solid cell culture technique. This article covers sterile technique, thawing and freezing, subculture and preparation of cells for grafting. Read more
Microbubbles for in vivo ultrasound mouse imaging are encapsulated, inert gas filled colloidal particles. Upon intravenous injection, they are used as ultrasound contrast agents. Read more
Inhalation anesthesia with isoflurane is quickly becoming the standard method of general anesthesia for rats and mice used in biomedical research and is imperative for in vivo imaging. Inhalation anesthetics, breathing circuits, gas scavenging systems and oxygen flow rate are discussed. Read more
Instrumentation and methods for gas anesthesia and dosing for injectable anesthetics such as xylazine/ketamine and avertin are described. Read more
