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FLUORO-RUBY USE GUIDE

PostPosted: Fri 11 Dec 2015 12:51
by milaP59
Hello,

please, Can I have the use guide/protocol of FLUORO-RUBY?
If you have also the product information sheet... thanks!

Best,

Mila

Re: FLUORO-RUBY USE GUIDE

PostPosted: Fri 11 Dec 2015 13:05
by Interchim
Hello Mila,

For the FLUORO-RUBY protocol :

A 10% solution of Fluoro-Ruby is made by dissolving 10 mg of Fluoro-Ruby dry powder in 100 µL of distilled water. It is then delivered via intracranial stereotaxic injection using a 1 uL Hamilton microsyringe to pressure inject the tracer. Injection volumes typically range from 0.02-0.1 µL and are gradually injected over a 10-15 minute interval. The animal is allowed to recover and is sacrificed typically 4-14 days later.

The animal is then perfused with neutral buffered formaldehyde (10% formalin or 4% paraformaldehyde in 0.1 M neutral phosphate buffer.) The brain is removed and postfixed for at least overnight in the same fixative solution plus 20% sucrose for cryo-protection. The brain is then cut on a freezing sliding microtome or cryostat into sections usually between 20 and 40 microns in thickness.

The tissue sections are mounted on gelatin coated slides and are air dried on a slide warmer at 50-60 degrees Centigrade for at least one half hour. The dry slides are then transferred to xylene clearing solution for at least one minute and then coverslipped with DPX mounting media. The slides can then be examined under an epifluorescent microscope using a filter suitable for visualizing TRITC (green light excitation).

If you need more, let us know.

Best regards,

Interchim

Re: FLUORO-RUBY USE GUIDE

PostPosted: Fri 11 Dec 2015 13:08
by Interchim
*About the product information :

Classification: Tetramethylrhodamine dextran-amine

Appearance: Pink to crimson colored powder

Solubility: Highly soluble in aqueous solutions

Purity: Thin layer chromatography using cellulose plates and a solvent system of acetonitrile and water (6:4) revealed no detectable free TRITC dye.

Substitution: 1 molecule TRITC per molecule of dextran-amine

Average Molecular Weight: 10,500

Excitation Peak: 540 nm

Emission peak: 600 nm

Filter system for visualizing: Standard TRITC filter

Storage: The dry powder can be stored in a tight container and may be stored at room temperature or in a commercial refrigerator without the frost free feature. Solutions should be tightly covered and refrigerated when not in use

Toxicity: Although the compound appears to be of low toxicity, it has not been extensively evaluated and therefore routine laboratory caution should be exercised. Not approved for human consumption.


Interchim