Merci Beaucoup Fred
Louis
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| De : "Fred E. Indig, Ph.D." <[hidden email]>@LISTSERV.BUFFALO.EDU
Envoyé par : Confocal Microscopy List <[hidden email]>
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cc : (ccc : Louis Villeneuve/ICM/Reg06/SSSS)
Objet : Re: DABCO-mounting medium
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| 2008-03-11 10:40
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Dear Louis,
I have used the following formula for over 10 years (Indig et al., Biochem J. 327:291-8, 1997) with excellent results:
(1) Dissolve in beaker (0.5 L for 100 mL solution):
DABCO (1,4-Diazabicyclo[2.2.2]-octane, Aldrich) 2.5% (w/v)
and Mowiol 4-88 (Calbiochem) 10% (w/v)
in 0.1M TRIS-HCl, pH 8.5 and 25% (w/v) glycerol
(2) This could take a while (hours)- use a heated magnetic stirrer at low speed, about 50 C.
(3) After all is dissolved, aliquot 0.25 mL into microtubes, store -20 C in the dark (good for well over one year).
(4) Before use, heat to 65 C for 10 min, let cool 1-2 min and use immediately on specimen. Discard remains. This mounting media will solidify overnight in the refrigerator (4 C, dark), but is not as hard as epoxy mounts, so handle gently.
(5) I measured the refractive index as R=1.4101 @ 22.0 C (water 1.33, oil 1.56)
Best Wishes,
Fred
Search the CONFOCAL archive at http://listserv.acsu.buffalo.edu/cgi-bin/wa?S1=confocal
Hi everyone,
I am looking for a recipe for mounting media with DABCO diluted in glycerol. The one I have is DABCO, 0.4% (diluted in PBS) mix 1:4 in glycerol. Also, where I could find a database for finding the refractive index of several mouting medium/media?
Thanks
Louis Villeneuve
Louis Villeneuve
Research Associate - Confocal Microscopy
Heart Montreal Institute- Research Center
5000 East Belanger
Montreal (Qc), Canada
H1T 1C8
514-376-3330 ext 3511
514-376-1355 (Fax)
[hidden email]
--
Fred E. Indig, Ph.D.
Head, Confocal Imaging Facility
Research Resources Branch
NIH Biomedical Research Center (BRC)
National Institute on Aging,
251 Bayview Boulevard,
Room #4B026
Baltimore, Maryland 21224
Tel. 410-558-8173
Fax 410-558-8236
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