Biology Reference
In-Depth Information
Fluo-3
A
1000
600
100
400
10
700
800
900
1000
1P
×
2
2P
200
0
700
800
900
1000
Wavelength (nm)
B
Rhod-2
1000
100
600
10
800
1000
1200
400
200
1P
×
2
2P
0
800
900
1000
1100
700
1200
Excitation wavelength (nm)
Fig. 8
Excitation spectra of Ca
2
þ
bound forms of (A) Fluo-3 and (B) Rhod-2. Emitted fluorescence
was collected at 500-650 nm (Fluo-3) and 550-650 nm (Rhod-2). Black line represents the single-photon
(1P) excitation spectra measured on a Perkin-Elmer spectrophotometer (2 nm slit width); the 1P
wavelength has been scaled by a factor of 2 (
2) to generate comparable wavelengths to two-photon
(2P) spectra. Gray line represents the 2P excitation spectrum of an equivalent concentration of dye (10
m
M); excitation light was provided by a Coherent Chameleon XP Ti:Sapphire laser attached to a Zeiss
510 upright laser scanning microscope. Laser power was altered at each wavelength to ensure equivalent
excitation power across the excitation wavelengths. Insets show spectra plotted on log scale.
