Fluoro Gold 英文名称: Fluoro Gold | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
型号:null 产品货号: FC10000 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
价格:请致电:010-57128832,18610462672 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
品牌: 美国 试剂级别: 超纯级 | |||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Fluorochrome 公司的荧光金通过专利认证,其Fluoro-Gold为全球独家供应。 最近,国内市场上出现假冒Fluorochrome的伪劣产品,导致老师的实验受到很大影响。 伪劣假冒产品特点:
Amygdala cells (40X) after Fluoro-Gold injection in the PVN. Antibody is at 1/50,000. Main Protocol 1. Background The use of Fluoro-Gold is essentially the same as other fluorescent tracers. The main difference is that Fluoro-Gold is more flexible in terms of post-injection survival times, concentration range, tissue treatment and compatibility with other histochemical techniques. 2. Storage and Shelf Life Dry Fluoro-Gold should be kept in a light tight closed container at 4 degrees Celsius. Stored properly, Fluorogold should have a shelf life exceeding one year. The dye in solution should also be kept in a light tight closed container at 4 degrees Celsius and should remain stable for at least six months. 3. Vehicle Fluoro-Gold can be dissolved in distilled water or 0.9% saline, or utilized as a suspension in 0.2M neutral phosphate buffer. 4. Dye Concentration Fluoro-Gold has been successfully used at concentrations ranging from 1-10%. Initially, a 4% concentration is advised. If undesirable necrosis occurs at the injection site, or labeling is too intense, reduce the concentration to a 2% solution. If you need to use more precise measurements, the molecular weight of Fluoro-Gold is 532.6 daltons. 5. Dye Administration A. Pressure Injection - This is probably the most frequently used mode of application. Volumes injected range from .05-1 µ l, typically .1-.2 µ l. B. Iontophoresis - Discrete, small injection sites result from 4-10 second pulsed iontophoretic (+5 to +10ua/10min) application. C. Crystal - A crystal of the tracer can be administered from the tip of a micro-pipette. 6. Post-0perative Survival Period Good retrograde labeling has been observed with periods ranging from two days to two months. Survival periods of three to five days are typical. Long survival periods enhance filling of distal processes without diffusion of the dye from the cell. 7. Fixation Almost any fixative, or no fixative, can be used, Phosphate neutral buffered saline containing 4% formaldehyde is frequently employed. Fixatives containing high concentrations of heavy metals (e.g. osmium, mercury) will quench the fluorescence, while high concentrations (over 1%) of glutaraldehyde may increase background fluorescence 8. Histochemical Processing Tissue containing Fluoro-Gold may be processed according to virtually any common histological technique. This includes cryostat sections of unfixed tissue (10 µ m), frozen sections of fixed tissue (20 µ m), and thin sections cut from tissue imbedded in either plastic (.2-4 µ m) or paraffin (3-10 µ m). Frozen sections of fixed tissue are most frequently used. 9. Combined Methods At this point of processing, sections may be further processed for a second marker such as autoradiography, HRP histochemistry, immunocytochemistry, a second fluorescent tracer, fluorescent counterstain, etc. 10. Mounting, Clearing and Coverslipping Sections are typically mounted on gelatin-coated slides, air-dried, immersed in xylene, and coverslipped with nonfluorescent DPX plastic mounting media. Sections may be dehydrated with graded alcohols, unless this is not compatible with a second tracer. If Fluoro-Gold is to be combined with fluorescence immunocytochemistry, then sections are air- dried and directly coverslipped with neutral buffered glycerine (1:2). 11. Examination and Photography Fluoro-Gold can be visualized with a fluorescence microscope using a wide band ultraviolet excitation filter. A gold color is emitted when tissue has been processed with neutral pH buffer, whereas a blue color is emitted when tissue is processed with acidic (e.g. pH 3.3) pH buffer. It can be photographed digitally or with film (use Ektachrome 200-400 ASA film for color prints and comparable speed film for black and white prints, for example Tri-X). Most exposure times range from 10-60 second exposures, depending on the objective magnification and the intensity of the label. Thirty (30) second exposures are about average. Multiple exposures may be exploited to simultaneously visualize Fluoro-Gold and another tracer. Thus, UV would be combined with bright field illumination to simultaneously locate Fluoro-Gold with HRP or silver grains in autoradiography. Similarly, blue light excitation can be combined to also visualize the green emission color of FITC, while green excitation light may be used to simultaneously observe the red emission color of propidium iodide, or ethidium bromide (a fluorescent counterstain). Additional Information Concerning the Use of Fluoro-Gold
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