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Kurihara, D., Mizuta, Y., Sato, Y. & Higashiyama, T. ClearSee: a rapid optical clearing reagent for whole-plant fluorescence imaging.
In addition, a number of other herbs and spices (dry samples and whole tissues) were analyzed subsequently using Visikol as clearing reagent with comparable results.
A 20 ml of clearing reagent (1/3 lactic acid+ 1/3 glycerol + 1/3 distilled water) were added to roots and roots were left to destain for two hours to overnight.
Congo red (a b-D-glucans stain) or Calcofluor white 0.1% solution (a chitin binding fluorochrome dye) added to the clearing reagent facilitate the visualization of fungal structures, but the latter requires use of a fluorescence microscope.
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Equipment Available for Service: Clearing reagents, dyes, and custom Multiphoton microscope.
Fluorescence-stained embryos and larvae cleared in Murray's reagent were scanned with a Leica SP5 confocal laser scanning microscope (Leica, Wetzlar, Germany).
From the point of view of laboratory workflow, the use of a uniform commercial reagent offers clear advantages in terms of staff time and avoids the need for revalidation of custom reagents.
Thereafter, embryos were washed in PBS for 1 h, dehydrated in a graded isopropanol series (70%, 85 %, 95 in PBS, and twice in 100% for 30 to 60 s each) and cleared in Murray's reagent (benzyl benzoate to benzyl alcohol, 2:1, v:v).
And then sections were rinsed through ascending grades of ethanol with a final three changes of 100% reagent grade ethanol, cleared in xylene and cover slipped Canadian balsam solution.
After cell seeding and treatment, the well plates were cleared and 20 μl of MTT reagent was added to each well.
In another experiment, the original infected MSC line was cleared of mycoplasma using Mynox® reagent (Minerva Biolabs, Germany) according to the manufacturer's protocol, and was confirmed negative by PCR.
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