The application of French VILBER Fusion series optical in vivo imaging

The application of French VILBER Fusion series optical in vivo imaging

At present, the main methods of animal and plant live imaging include structural imaging and functional imaging. Structural imaging mainly uses ultrasound, computed tomography (CT), nuclear magnetic resonance and other methods, while functional imaging mainly uses optical imaging. Optical imaging mainly uses bioluminescence and fluorescence technologies to carry out biomarkers in living animals and plants, and uses imaging systems to detect the development of molecules and cells in labeled animals and plants, which can be used for cancer and drug research. , Virology and gene therapy, apoptosis, protein interaction and transgenic animal models, etc.

The application of French VILBER Fusion series optical in vivo imaging

In the process of optical live imaging, there are still some challenges. First, due to the influence of the position of the probe or dye in the animal body, the deeper the position, the more difficult it is for the signal to penetrate the animal skin and be difficult to detect; second, the selected excitation light source wavelength and dye emission wavelength are shorter, the same Can't penetrate the skin well; finally, because the probe or dye is usually injected through the tail vein, the probe will be diluted by the animal's blood and tissues, and the signal in the body is relatively weak. Therefore, the selected probe or package The nanomaterials to be dyed are preferably targeted.

In response to the above problems, the Fusion FX7 multi-function imaging system developed and produced by VILBER in France adopts a four-stage semiconductor cooling CCD, which can be as low as -67 ℃ relative to room temperature, and the dark current is only 0.0002e / p / s, which greatly reduces the background Noise; 1.2inch large-area CCD sensor with f0.84 large-aperture fixed-focus lens minimizes the loss of optical signals and improves the detection sensitivity, even if there are few photons in animals and plants to detect the signal. In the process of in vivo imaging, the wavelength and frequency of the excitation light source have a great influence on the excitation of the dye. The longer the wavelength, the higher the frequency, the more it can penetrate animal skin tissues and improve the incidence of excitation light. VILBER Fusion FX7's unique Pulsed light technology increases the emission frequency of the light source, thereby increasing the amount of incident light per unit time. In addition, Fusion FX7 IR Spectra has 640nm red light and 740nm far red light excitation light source, which can effectively penetrate animals Skin tissues, after activating the dye, acquire more emitted light signals.

The application of French VILBER Fusion series optical in vivo imaging

During the in vivo imaging experiment, you should also pay attention to the following points: First, try to choose fluorescent dyes with long excitation and emission wavelengths, such as CY5, CY5.5, Qdot 705, or Alex Fluor 700. If you use fluorescent probes, Try to make sure that the probe performance is stable and reliable in vitro, and determine the emission wavelength or intensity. You can also use blood and other biological fluids to dissolve the probe to observe its changes. Secondly, since the animal itself will have different autofluorescence in different parts, and under different excitation light conditions, the autofluorescence will also be significantly different. Therefore, if mice are used for experiments, try to select nude mice as much as possible to avoid fluorescence interference from the fur of the mice. Furthermore, the nanomaterials or drugs and the dyes must be thoroughly mixed so that the drugs and the like are sufficiently coated with the dyes to increase the signal strength and also prevent the occurrence of false positives.

Finally, a contrast should be set up for in vivo imaging. After the spectrum is determined by the positive control, the negative control group is split under the split conditions for spectral separation. If the conditions are suitable, the negative control should have no corresponding signal. The location of the signal can also be determined based on existing knowledge and time changes.

When using VILBER Fusion FX7 for in vivo imaging, the recommended settings are as follows:

If it is biological self-luminescence, select Chemiluminescence imaging mode, set to f0.84 aperture, automatically or manually calculate the optimal exposure time, you can also select Video mode for continuous image capture to obtain the best imaging.

The application of French VILBER Fusion series optical in vivo imaging

If you choose a fluorescent dye, choose the Fluorescence imaging mode, choose the appropriate light source and filter according to the selected dye, the aperture can be set to f1.2, and it can be adjusted according to the signal strength. Select the manual mode to increase and decrease the exposure time according to the signal strength.

After the imaging is completed, pseudo colors can also be added to distinguish the strength of the signal with different colors.

Applications:

The following is the image obtained in the VILBER Fusion FX7 Spectra series imaging system of the mouse in vivo drug metabolism analysis experiment conducted by the Affiliated Hospital of Qingdao University School of Medicine and China Ocean University:

FITC dye replaces drug-coated nanomaterials and is injected into mice by gavage to observe the distribution of drugs (signals) in the body during different periods of time. The following is the signal distribution of 2h \ 4h \ 6h after intragastric administration.

The application of French VILBER Fusion series optical in vivo imaging

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