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华拓生物为Fluorochrome公司国内授权代理销售Fluoro Gold

放大字体  缩小字体 发布日期:2014-03-28  浏览次数:5

 

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:
1,供应公司声称自己为国外很多品牌的一级代理,但均无授权证明的。
2,供应产品信息不全,宣传推广只宣传国外某个品牌,没有该品牌的详细产品信息,并在其他网站也找不到相应代理信息。如有的公司只做个品牌的百度推广,但查询不到任何具体产品信息的。
3,声称荧光金大量现货,各种规格均有销售。如:10mg,30mg (Fluorochrome公司荧光金无法分装,只有20mg、50mg、100mg、150mg、200mg的包装规格) 
4,价格低廉,均低于市场均价,质量差。
 

公司近期收到一些水货及假货的投诉,特此声明,不论假货或水货,因进货渠道不明,运输条件不合规格,产品效用不明,一律没有产品保证,绝不退款或换货。 
深圳市豪地华拓生物科技有限公司 Fluorochrome公司在中国地区的指定授权代理。 为保障您的实验结果,我们郑重建议各位客户及经销商在选购Fluorochrome产品时,务必确认产品为正规来源。

 
美国Fluorochrome《荧光金》
Fluoro-Gold
Use Guide and Protocol
 
US Patent No. 4,716,905
UK Patent No. 2,181,545

 

FC10000 荧光金 Fluoro-Gold 20mg   Fluorochrome
FC10001 荧光金 Fluoro-Gold 50mg   Fluorochrome
FC10002 荧光金 Fluoro-Gold 100mg   Fluorochrome
FC10003 荧光金 Fluoro-Gold 150mg   Fluorochrome
FC10004 荧光金 Fluoro-Gold 200mg   Fluorochrome
FC20001 荧光金抗体 Antibody to Fluoro-Gold 0.1ml×1   Fluorochrome
FC20002 荧光金抗体 Antibody to Fluoro-Gold 0.1ml×2   Fluorochrome
FC20003 荧光金抗体 Antibody to Fluoro-Gold 0.1ml×3   Fluorochrome
FC30001 红色荧光金 Fluoro-Ruby 30mg   Fluorochrome
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 me-tals (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 (10µ 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 ab0ut 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

 

Vehicle For pressure injections through a microsyringe or micropipette, Fluoro-Gold should be dissolved in distilled water or .9% saline. Fluoro-Gold may also be utilized as a suspension in .2M neutral phosphate buffer, however, the suspended particles may clog a fine micropipette tip so distilled water or .9% saline is the preferred vehicle. For iontophoresis, a 1% Fluoro-Gold solution is made up in .1M acetate buffer (pH=3.3). Well-cleaned (95% ETOH, water) glass micropipettes should have tips of 10-20 µ m. Optimal iontophoresis parameters are +1 to +5u amps delivered with pulsed current (4-10 seconds on, 4-10 seconds off) over a 10-20 minute period. Injection Sites Virtually any central or peripheral nervous system structure can be injected with Fluoro-Gold for analysis of retrograde transport. In the peripheral nervous system, ganglia and peripheral targets can be studied. For studies of peripheral nerve, the nerve should be cut or damaged and either dipped in, or injected with, aq 5% solution of Fluoro-Gold. Since Fluoro-Gold is not significantly taken up by intact fibers of passage, the fibers must be cut or severely damaged for uptake of the dye to occur. Transport and Survival TimeFluoro-Gold is used as a retrograde axonal tracer, although orthograde axonal transport does occur. The survival time should be varied (especially to very short survival times of 12 hours - 2 days) to maximize orthograde transport in the specific neuronal system under study. For retrograde transport, the survival times should be varied from 4 days to 14 days. Seven to 10 days works for most systems, although long pathways (e.g., spinal cord to brainstem) and pathways in large mammals (e.g., cats, m0nkeys) may require longer survival times (e.g., 14 days). In addition, since Fluoro-Gold remains fast within retrogradely labeled neurons, survival times of several months will also produce excellent results. For iontophoresis, a 2-5 day survival time is recommended. It is estimated that transport occurs at ab0ut 2 cm per day for mammals; it is slower for cold-blooded animals. Tissue Processing Tissue processing is covered in detail in the use guide and in the original publication ( Schmued and Fallon, 1986, Brain Research 377:147-154 ). Since Fluoro-Gold is stable in many solvents and remains fast within retrogradely labeled neurons, its use is compatible with many histochemical techniques. It can be used with other retrograde tracers, immunofluorescence, PAP and ABC immunocytochemistry, HRP histochemistry, autoradiography, counterstains (ethidium bromide is the preferred fluorescent counterstain), paraffin embedding and plastic embedding. However, if tissue is unfixed, additional processing of tissue in aqueous solutions for over an hour or two will result in loss of Fluoro-Gold fluorescence from labeled neurons. Fluoro-Gold may be useful in electron microscopy. Fluoro-Gold can be used in a brain which has been sectioned and transferred to phosphate buffer. Sections are typically mounted on gelatin-coated slides, air dried, immersed in xylene and coverslipped with DPX plastic mounting media (FLUKA Chemical Corp., 255 Oser Avenue, Hauppauge, New York, 11788, Catalog #44581). Tissue may also be viewed on slides without further processing, can be run through graded alcohols for dehydration, or, for immunocytochemistry, the sections can be air dried and directly coverslipped with neutral buffered glycerine (1:2). Examination and Photography Fluoro-Gold is visualized with a fluorescence microscope using a wide band ultraviolet (UV) excitation filter. Use the same filter pack you would for other fluorescent retrograde tracers excited under wide band UV (e.g., True Blue, Fast Blue, Nuclear Yellow), such as the Leitz Ploem filter system A (Wide Band UV, Excitation filter BP 340-380), Mirror RKP 400, Barrier Filter LP 430). Objectives should be made especially for fluorescence microscopy (such as that made by Zeiss) glycerine, or water. Since plastic does absorb UV light, it is not advised to view through plastic petri dishes, etc. Recommended films are T-Max (Kodak, black & white) and Ektachrome 200 (Kodak, color slides). Exposure times usually vary from 20 seconds to 1.5 minutes. Chemical Analysis

Quality Expected Result Actual Result
Appearance A golden-yellow, hygroscopic, crystalline powder A bright-yellow powder
Odor None None
Solution 20 ml of a 5% w/v aqueous solution should be clean, clear and almost free from suspended matter, and should have not more than a very slight odor Passes Test
pH of a 1% Solution Between 4.0 and 5.5 at 25 degrees Celsius 4.6
Spectral characteristics The spectral characteristics of Fluoro-Gold vary with pH A 0.1% solution in distilled water has a pH of 4.5 and excitation peak of 414 nm and emission peak of 541 nm
Fluoro-Gold bound to membranes at a physiological pH of 7.4 has an excitation band of 350 to 395 nm and an emission band of 530 to 600 nm
Chloride Not more than 0.035% 0.017%
Sulfate Not more than 0.1% Less than 0.05%
Sulfated ash Not more than 0.1% Negligible
Heavy me-tals Not more than 10 p.p.m Less than 10 p.p.m
Selenium Not more than 30 p.p.m Less than 10 p.p.m.
Loss on drying Not more than 1.0% after 3 hours in vacuo at 60 degrees Celsius 0.1%
Assay Between 95.0 and 105.0% calculated with reference to the dried material 99.2%
Notice: The original and only true Fluoro-Gold (Fluorogold) is produced by Fluorochrome, LLC and marketed by Fluorochrome, LLC and Histo-Chem Inc.
深圳市华拓生物科技有限公司( www.otwobiotech.com

 

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