Re: Nuclear Dyes at 405nm?

Posted by Boswell, Carl A - (cboswell) on
URL: http://confocal-microscopy-list.275.s1.nabble.com/Nuclear-Dyes-at-405nm-tp3144876p3145707.html

Griffin,

 

We have excellent nuclear imaging with DAPI using our 405 laser, but it has not been without issues.  First off, I am curious as to the imaging system you are using, and has DAPI ever looked good on it?  I ask this because our Zeiss LSM 510 Meta has had troubles with 1) the AOM controlling the 405 input, 2) the electronics in the 405 system and 3) the laser itself.  Once each of these was dealt with (over the last 2 years), we returned to blissful DAPI imaging.  Currently all parts are holding up well and all users are quite happy with their very bright DAPI images.  

 

Aside from the above mentioned items, alignment is one of the first things to look at.

 

C

 

Carl A. Boswell, Ph.D.
Molecular and Cellular Biology
University of Arizona
520-954-7053
FAX [phone]

----- Original Message -----

From: [hidden email]

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Sent: Tuesday, June 23, 2009 1:30 PM

Subject: Nuclear Dyes at 405nm?

Dear Listserv,

 

I was wondering if anyone had better results with DAPI @ 405nm than I have.  It seems…off.  I know it is only getting excited @ 25%, but I know another group that gets good images at the same wavelength.  The staining has been uneven with DAPI, but with Topro-3 in the 638nm range, the staining has been beautiful with clearly defined nucleoli.  Does anybody use Hoechst 34580?    Are there any other alternatives to DAPI @ the 405nm range?

 

Thanks in advance,

 

-Griffin