🚀 go-pugleaf

On Mon, 29 May 1995, Jeff Craig wrote:

> On the subject of dual label fluorescence, I have performed a little
> comparison of absorbtion and emission spectra from various fluorochromes and
> worked out which 2 would be compatible for the microscopes/filters we have.
> I copied the spectra from the following molecules onto transparencies and
> overlayed combinations together with the absorbtion/emission details of each
> microscope. For our BioRad MRC 600 confocal, which excites at 514nm and emits
> at 525-555nm and >600nm, the following combinations would theoretically have
> no bleed-through:
> FITC and TexasRed (or Propidium Iodide DNA stain)
>....................................

While Jeff's theoretical approach seems very reasonable, life shows that
FITC spills over not only to Tx Red channel but even to Cy5 channel.
Theory versus practice - here we go again, Lenin.

Dr. Michal Opas
Department of Anatomy & Cell Biology
University of Toronto
Medical Sciences Building
Toronto, Ontario, M5S 1A8 Canada

tel 416 978-8947
fax 416 978-3954
e-mail m.opas@utoronto.ca