Mouse live imaging experiment
mouse live imaging experiment
Small animal biological imaging mainly uses biological lighting (Bioluminescence) Fluorescence (Fluorescence). Bioluminescence is labeled cells or DNA with luciferase genes, while fluorescence techniques are labeled with fluorescent reporting groups (GFP, RFP, CYT and DYES, etc.).
Technical application (mouse live imaging experiment)
The development process of disease or cancer can be observed by the vivo imaging system, and the reaction produced by drug treatment can be observed, and the reaction produced by drug treatment is used for viral. Study, construct transgenic animal model, siRNA study, stem cell research, protein interaction research, and cell in vitro testing. The specific application is as follows:
(1)
Cancer and anti-cancer drug research
Directly measure various cancer models The growth and metastasis of tumors, and can perform real-time observation and evaluation of cancer the treatment of cancer cells. The living bioremor imaging can quantify the overall nomography, metastatic tumor and self-tumor of the mice in the absence of injury.
Immunology and stem cell studies
Transplant luciferase labeled hematopoietic stem cells into spleen and bone marrow, which can be used in real time to observe an early incidents of stem cell hematopoietic processes in vivo animals. And kinetic changes. Studies have shown that mice with bioluminescent labeling genes, the effect of detecting radiation and chemical drug treatment, looking for complex cellular mechanisms in tumor bone marrow metastasis and anti-tumor immunotherapy. Application visible photoreceptive imaging principle marked cells, establishing an animal model, which can effectively observe the same group of animals, saving animal samples, and can also get a pathogenic pathway and resistance protein expression in the immune system more quickly. Change.
When the fluorescentase and inhibitory polypeptide are expressed in mammalian cells in a fusion protein form, the resulting fusion protein-free luciferase activity, cell It is not possible to illuminate, while when the cells are apoptosis, activated caspase-3 cut off the inhibitory protein in a specific identification site, restores luciferase activity, resulting in a luminous phenomenon, thereby useful in observing apoptosis in the living animal. Related events .
2. Mark Virus
Tried a luciferase gene-labeled HSV-1 virus as an example, observedHSV-1 viruses to the process of the liver, lung, spleen and lymph nodes and viruses from the blood system into the nervous system.A variety of viruses, adenovirus, adenoga-free viruses, slow viruses, hepatitis B viruses, etc., have been labeled by luciferase, used to observe the infection process of the virus on the body.
(2) Gene therapy
gene therapy includes transmitting one or more genes of interest to the target cells in vivo.The phosphoridase gene can be applied as a reporter for the construction of the carrier, and the object gene can be observed whether the gene can continue to be efficient and tissue-specific expression in the test animal.This non-invasive manner has the advantage of easy preparation, low toxicity and mild immune response.The luciferase gene can also be inserted into the DNA molecule wrapped in the liposome to observe the DNA transport and gene therapy of the liposome as the carrier.