Re: Hoescht 33342 and UV Excitation with 325nm HeCd Laser

From: Nigel Blackhall (Nigel.Blackhall@nottingham.ac.uk)
Date: Thu Oct 12 2000 - 05:28:47 EST


Dear all

Jim Houston stated:

The optimum excitation of 33342 is with the 351 line of the argon laser.
With emmision around 460+/-.
When using the 325 line of the HeCd can I expected a lower emmision output?

I would like to run the SP population protocol that was published by Stroms
et al. in Blood 15 September 2000-Vol 96.
They indicated using the 351 line in combination with a 450DF20 filter for
the Blue and a LP675 for the Red.

The Question???  Will the 450/20 work with the 325 line or do I need expect
it to shift down.??

Any help is appreciated.

***********************
Reply:

Emission from fluorochromes is generally independent of excitation wavelength (as long as its exciting the same electron orbits).  Therefore keep the 450DF20 and LP675.  However, you will need to adjust the filter which sends laser light to the 90° (wide angle) scatter detector.

Regards,

Nigel Blackhall
School of Biosciences
Plant Science Division
University of Nottingham
Biological Sciences Building
University Park
Nottingham NG7 2RD
UK
Tel: +44 (0)115 9515151 ext 18501
Fax: +44 (0)115 9513298
Email: Nigel.Blackhall@Nottingham.ac.uk
http://plantsci.nottingham.ac.uk
http://www.nottingham.ac.uk



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