Re: microbead calibration to set psf?

Posted by Zoltan on
URL: http://confocal-microscopy-list.275.s1.nabble.com/microbead-calibration-to-set-psf-tp2560403p2563282.html

David,

 

Thermo still lists the beads:

 

http://www.thermo.com/com/cda/product/detail/1,,10138303,00.html

 

Just checked it, is that what you need?

 

Zoltan

 

 

On Tue, Mar 31, 2009 at 2:56 PM, David Knecht <[hidden email]> wrote:

I am looking for some very bright 100nm beads.  I tried to look for the beads you mention and it looks like Duke Scientific no longer exists and is now part of hte Thermo conglomerate.  The closest thing I found to what you describe is the Europium chelate 100nm beads.  Is that what you were referrring to?  THey provide very little data on fluorescence properties and they are unusual in apparently being broad excitation spectrum but there is no emission spectrum in their literature I can find.  Since they fluoresce maximally at 613nm it would seem they would be a poor fit for most FITC, TRITC, CY3 or CY5 type filter sets.  Can you clarify your experience with them?  Dave

On Mar 31, 2009, at 3:36 AM, Mariette P.C. Kemner - van de Corput wrote:

I have good experience with fluorescent beads from Duke scientific. The
100nm beads are very bright and are perfect for determining the PSF. I use
the red, green and blue 100nm beads to determine the PSF in three channels
and to deconvolved my 3 colour FISH images.
I have trouble finding the 100nm and 200nm TetraSpeck beads on my slide
beacuse they are rather weak in intensity. So that's why I switched to the
single coloured 100nm beads from DS. The 500nm TetraSpeck beads are very
good and I use them to determine the chromatic shift in 4 channels. Once
made, bead slides can be used for a long period of time.

Mariette

____________________________________________

Dr. M.P.C. Kemner-van de Corput,
____________________________________________

MGC - Dept. of Cell Biology & Genetics
Erasmus Medical Center
Dr. Molewaterplein 50, 3015 GE Rotterdam
POB 2040, 3000 CA Rotterdam, The Netherlands

Office: H-Ee751; tel: [phone]
Lab: H-Ee710; tel lab: +31 10 704.3315
tel secr: +31 10 704.3169
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http://www.thesis.kemner.biz/
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Op Di, 31 maart, 2009 1:59 am, schreef Matiar Jafari:

hey again,

I'm trying to find out how and what the best way to setup my own psf

is in AutoQuant.  I know i need microbeads and im wondering what brand

and what product i need and then how i need to shoot it and load it.

Also what diameter does it need to be.  A cat# would be great too also

i don't know if it matters on what im shooting but just to put it out

there im shooting gfp slices.  Sorry if this is vague. Any help would

be greatly appreciated.  Thanks in advance

Thank You

--

Matiar Jafari

Dr. David Knecht    

Department of Molecular and Cell Biology

Co-head Flow Cytometry and Confocal Microscopy Facility

U-3125

91 N. Eagleville Rd.

University of Connecticut

Storrs, CT 06269

860-486-2200

860-486-4331 (fax)

--

Zoltan Cseresnyes
Facility manager, Imaging Suite