Iron Man User Guide v. 1.4
15
-
You need to know the
excitation and emission spectra
for all the dyes/fluorophores/probes you
will be using. This can readily be viewed in various Spectrum viewers on the web (list of which is on
the LMCF Website Resources tab at the bottom. Life Technologies’ SpectraViewer is a
straighforward and reasonably complete one. Excitation spectra are shown as dotted lines and
emission spectra are solid
-
Next, figure out which
lasers
are appropriate. Available lines: Argon/2 –
458
,
477
,
488
,
514
;
HeNe
–
543;
HeNe –
633
;
Diode –
405/30
.
In this example, we’ll use the 405 (DAPI), 488 (FITC) and 543
(Rhodamine). Shown here are the excitation spectra and the laser lines we’ve selected.
