Overview of gel imaging

Gel imaging definition
Gel imaging is a non-chemiluminescence imaging analysis of different staining of dna/rna/protein and other gel electrophoresis (such as eb, Coomassie blue, silver stain, sybr green) and microplates and plates. Gel imaging systems can be applied to routine engineering studies such as molecular weight calculation, density scanning, density quantification, and PCR quantification.

Principle of gel imaging system

The mobility of the sample on the electrophoresis gel or other carrier is not the same. A qualitative analysis of the unknown sample is performed by comparison with a standard or other alternative standard. This is the basis for the qualitative analysis of image analysis systems. According to the location of the unknown sample in the map, it can be qualitatively analyzed to determine its composition and properties. The sample partially absorbs the projected or reflected light, so that the optical density of the sample strip on the image obtained by the photographing is different. The optical density is linear with respect to the concentration or mass of the sample. Depending on the optical density of the unknown sample, the concentration or mass of the unknown sample can be obtained by comparing the optical density of the sample strips of known concentration. This is the basis for the quantitative analysis of image analysis systems. The use of the latest technology of ultraviolet transmission light source and white light transmission source to make the light distribution more uniform, to minimize the impact of uneven optical density on the results.

Gel imaging application range

In general, gel imaging can be applied to: gel imaging systems can be used for qualitative analysis of separation and purification results of other biomolecules such as proteins, nucleic acids, peptides, amino acids, and polyamino acids. (1) Molecular weight quantification For the commonly used DNA film, the molecular weight quantification function is used to automatically generate a fitting curve by annotating the known molecular weight of the DNA Marker band on the gel, and measuring the molecular weight of the unknown band by it. The results obtained by this method are much more accurate than the visual observations. (2) Density quantification A commonly used method for determining the concentration of DNA (deoxyribonucleic acid) and RNA (ribonucleic acid) is the ultraviolet absorption method, but it can only measure the total nucleotide concentration in the sample, and cannot distinguish the concentration of each length fragment. Using a gel imaging system and software, after density calibration of a standard strip of known DNA content on a DNA film, it is convenient to click on other unknown bands, based on the density of the known bands. The amount of unknown DNA can be obtained. This method is also applicable to the determination of the concentration of the PA GE protein strip. (3) Density scanning In molecular biology and bioengineering research, the most commonly used is the calculation of the percentage of protein expression products in the total bacterial protein. The traditional approach is to use a dedicated density scan, but this can be done using bioanalytical software in conjunction with a scanner that is routinely equipped in the lab or a gel that is directly illuminated with white light. (4) PCR quantification PCR quantification mainly means that if the bands amplified by the PCR experiment are not one, the software can calculate the relative percentage of each strip to the total strip. For this function, it is similar to density scanning, but it is not the same in principle. PCR quantification is to quantify the relative density of selected bands and calculate the percentage of the total. When the density is scanned, a longitudinal scan curve is generated and integrated into the selected area.

Gel imaging type

(1) Ordinary gel imaging analysis system: image collection and qualitative analysis of protein gel electrophoresis gel and DNA gel samples, including: EB, SYBR Green, SYBR Gold, Texas Red, GelStar, Fluoroscecin, Nucleic acid monitoring of Radiant Red and the like; and Coomassie Blue, SYPRO Orange, various stained protein gels such as test dyes. (or UV, EB and colored and visible sample imaging); (2) Chemiluminescence imaging analysis system: imaging range covers UV, EB, chemiluminescence, ultraviolet-fluorescence, colored and visible sample imaging; (3) Multicolor fluorescence imaging analysis system: imaging range covers UV, EB, chemiluminescence, multicolor fluorescent fluorescence, colored and visible sample imaging; (4) Multi-function in vivo imaging analysis system: UV, EB, chemiluminescence, multi-color fluorescent fluorescence, colored and visible sample imaging and isolated tissues and small animals, and large animals.

Gel imaging manufacturer

At present, there are many gel imaging manufacturers, and the common gel imaging on the market is: American Western Union (sim) brand gel imaging, Japanese astron gel imaging and so on.

Gel imaging features

Take the American West International (sim) brand Bio-Best 140M gel imaging as an example: High resolution pixels: 1.4 million, 2 million, 300 high pixel professional digital CCD camera, the analysis of images is clearer and the resolution is higher. Push-pull workbench: Easy to handle, clean, easy to experiment and humanized design. Unique fan and closed duct design: The UV workbench and the main unit are equipped with an exhaust fan. The side wall of the main unit has SIM's patented closed air exhaust passage, which effectively reduces the indoor temperature. The maximum procedure reduces the impact on the palm and prevents the temperature from being too high to make the glue disperse too fast. Seal design: It protects the operator from radiation that leaks out ultraviolet light. Advanced black box technology for safe operation. A variety of light sources are available: The four applications of the 312nm UV, 254nm UV, 365nm UV and white light that can be controlled by the touch key extend the range of applications. Push-pull workbench: Easy to handle, clean, easy to experiment and humanized design. Unique fan and closed duct design: The UV workbench and the main unit are equipped with an exhaust fan. The side wall of the main unit has a SIM exclusive patented closed exhaust passage to effectively reduce the indoor temperature. The maximum program reduces the influence on the sample and avoids the test being too high to make the glue disperse too fast. Seal design: It protects the operator from radiation that leaks out ultraviolet light. Advanced black box technology, safe operation: The sealing effect is good to ensure the acquisition of pictures and the legend of UV.

Gel imaging general procedure

1. Turn on the gel imaging system switch. 2. Turn on the computer and the system automatically turns on and enters the imaging software. 3. Open the front panel of the gel imaging system and choose to use the UV or white light transmission source to place the corresponding light source in place. 4. Place the sample on the sample stage of the transmitted light source. 5. Select the Upper white light source in the imaging operation interface and click the green (immediate imaging) button.

common problem

1. Is the higher the pixel, the better the product? Pixels are to be viewed for imaging devices. For example, the pixels of a digital camera and a CCD are not comparable, and the quality of the CCD itself is more important than the pixel level. The most important indicator for the same level of CCD is its size, which is the CCD size. The larger the size, the more its own value grows geometrically. 2. What are the main uses of UV light source for accessories? The UV-reflecting lamp source is not widely used, and is mainly used for providing imaging services such as paper chromatography for non-transparent materials. Because the more commonly used transparent agarose gel and polyacrylamide gel, the transmission source can be fully satisfied, so the UV-reflecting light source as an option is not included in the standard. 3. Can I cut the gel directly on the gel image? TOCAN gel imaging can, if you need to directly cut the glue, first press the observation window in the middle of the system, open the observation window with imported anti-UV glass, and then open the small door designed for the tapping on the left and right sides, you can cut the glue. operating. 4. How to adjust the focus focus aperture to get high quality pictures?

The three most important tunable parameters are focal focus and aperture. Among them, the focus adjusts the sharpness, the focus adjusts the image size, and the aperture adjusts the brightness. Generally, the aperture is adjusted to a suitable brightness, then the focus is adjusted to maximize the image, and finally the focal length is adjusted. In the case of a large image, the focal length is relatively easy to adjust to the clearest state, and then the image is scaled to an appropriate size for imaging. Just fine.

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