Semrock Master Catalog 2018

Fluorophores
Single-band
Sets
Multiband
Sets
Cubes Laser
Sets
NLO
Filters
BrightLine ® FRET Single-band Sets
These filter sets offer our simplest solution for dual-wavelength FRET imaging.
Also see our multiband “Sedat” filter sets for high-performance FRET imaging starting on page 28.
Filter Set /
Primary Fluorophores
FRET-BFP/GFP-B
Blue: BFP, DAPI, Hoechst, Alexa Fluor ® 350
Green: GFP, (EGFP), FITC, Cy2 , Alexa Fluor ® 488
FRET-CFP/YFP-C
Cyan: CFP, CyPet, AmCyan
Yellow: YFP, YPet, Venus
FRET-GFP/RFP-D
Green: GFP, (EGFP), FITC, Cy2 , Alexa Fluor ® 488
Red: mRFP1, mCherry, mStrawberry, dTomato,
DsRed, TRITC, Cy3
FRET-CY3/CY5-A
Yellow: Cy3 , Alexa Fluor ® 555
Red: Cy5 , Alexa Fluor ® 647
Excitation
(CWL/BW)
Emission
(CWL/BW)
387/11 Em1: 447/60
Em2: 520/35
438/24 Em1: 483/32
Em2: 542/27
472/30 Em1: 520/35
Em2: 641/75
531/40 Em1: 593/40
Em2: 676/29
Dichroic
(Edge)
Filter Set Part Numbers /
–ZERO Set Part Numbers
409 nm FRET-BFP/GFP-B-000
FRET-BFP/GFP-B-000-ZERO
458 nm FRET-CFP/YFP-C-000
FRET-CFP/YFP-C-000-ZERO
495 nm FRET-GFP/RFP-D-000
FRET-GFP/RFP-D-000-ZERO
562 nm FRET-CY3/CY5-A-000
FRET-CY3/CY5-A-000-ZERO
Filter Set and imaging dichroic options for common FRET fluorophore pairs
Start with a FRET
fluorophore pair
For imaging with emission
filter wheel only, using a
FRET single-band filter set
For imaging utilizing
emission signal splitting,
add an image
splitting dichroic
BFP/GFP FRET-BFP/GFP-B FRET-BFP/GFP-B
FF484-FDi01-25x36
CFP/YFP FRET-CFP/YFP-C FRET-CFP/YFP-C
FF509-FDi01-25x36
GFP/RFP FRET-GFP/RFP-C FRET-GFP/RFP-C
FF580-FDi01-25x36
Cy3/Cy5 FRET-Cy3/Cy5-A FRET-Cy3/Cy5-A
FF662-FDi01-25x36
See spectra graphs and ASCII data for these filter sets at
www.semrock.com/FRET-SingleBand.aspx
Base Price /
–ZERO
Price
$1125
$1224
$1125
$1224
$1125
$1224
$1125
$1224
Filter Specifications on page 34
“–ZERO” denotes zero pixel shift performance (see page 36)
For imaging with excitation
and emission filter
wheels, “Sedat” multiband
filter sets
DA/FI-2X2M-B
CFP/YFP-2X2M-B
FITC/TxRed-2X2M-B
Cy3/Cy5-2X2M-B
For imaging with
excitation filter wheel
and emission signal splitting,
add image splitting dichroic
DA/FI-2X2M-B
FF484-FDi01-25x36
CFP/YFP-2X2M-B
FF509-FDi01-25x36
FITC/TxRed-2X2M-B
FF580-FDi01-25x36
Cy3/Cy5-2X2M-B
FF662-FDi01-25x36
Image splitting dichroics are listed on page 64
CUBES PAGE 35
Individual
Filters
Dichroic
Beamsplitters
Tunable
Filters
TECHNICAL NOTE
Fluorescence Resonance Energy Transfer (FRET)
Fluorescence (or Förster) Resonance Energy Transfer (FRET) is a powerful
technique for characterizing distance-dependent interactions on a molecular
scale. FRET starts with the excitation of a “donor” fluorophore molecule by
incident light within its absorption spectrum. If another fluorophore molecule
(the “acceptor”) is in close proximity to the donor and has an absorption
spectrum that overlaps the donor emission spectrum, nonradiative energy
transfer may occur between donor and acceptor. For example, CFP and YFP
support a strong FRET interaction. FRET can measure distances on the order
of the “Förster distance” – typically 20 to 90 Å. This length scale is far below
the Rayleigh-criterion resolution limit of an optical microscope (about 2500 Å
for visible light and high numerical aperture), thus illustrating the power of
FRET for measuring extremely small distance interactions.
100
90
80
70
60
50
CFP Abs
CFP Em
40
30
20
10
0
400 450 500 550
Wavelength (nm)
The figure at the right shows as an example the CFP and YFP absorption and emission spectra, along with the transmission
spectra of the filters in the FRET-CFP/YFP-A set. Sets like this one and those listed above are optimized for the FRET-cube
method of imaging.
Transmission (%)
Exciter
Emitters
Dichroic
YFP Abs
YFP Em
600
More
22