Four Exciters

 

LED-based light engines are steadily replacing traditional mercury, metal-halide and xenon light sources in fluorescence microscopy.  As the spectral output of LED-based light sources differs from that of traditional sources, optical filter sets which were designed to take advantage of the peaks of the traditional sources, may now be sub-optimal for use with LED-based light engines.  Semrock is now offering eleven initial sets in single-band, full & Pinkel multiband configurations designed to be spectrally aligned with the excitation output of the most popular LED light engines on the market today.

 

The LED-DA/FI/TR/Cy5-4X-A is a “Pinkel” quad-band set designed for imaging of a sample that is simultaneously labeled with DAPI, FITC, TRITC, and Cy5 (or other key fluorophores – see complete list on the set page). This state-of-the-art set represents a break-through in quad-band performance, and has been optimized for popular LED-based light engines. This set has been designed to deliver significant fluorescence signal improvements for LED-based light engines compared with traditional sets designed for standard broadband light sources such as mercury or xenon arc lamps. The complete set is comprised of a quad-band beamsplitter and a quad-band emitter (these can be offered assembled in a microscope cube), plus four “no burn-out” single-band exciters that are intended to be mounted in the LED-based light engine, or in an exciter filter wheel or slider.

The part numbering of these new filter sets follows the convention LED-Fluor1/Fluor2/Fluor3/….- and will be available in Single-Band as well as Full Multiband, Pinkel, and Sedat configurations. The full line-up of Semrock’s LED-based light engine filter sets can also be found on our product sheet.

 

DA/FI/TR/Cy5-4X-B
LED-DA/FI/TR/Cy5-4X-A
DAPI-Exciter FF01-387/11 FF01-392/23
FITC-Exciter FF02-485/20 FF01-474/27
TRITC-Exciter FF01-560/25 FF01-554/23
Cy5-Exciter FF01-650/13 FF01-635/18
Dichroic FF410/504/582/669-Di01 FF409/493/573/652-Di01
Emitter FF01-440/521/607/700 FF01-432/515/595/730

 

SearchLight modeling tool shows the LED-DA/FI/TR/Cy5-4X-A filter set optimized for LED-based light engines performs as good as or considerably better in all channels compared with our standard broadband illumination source “Pinkel” set, DA/FI/TR/Cy5-4X-B, and delivers a significant performance improvement specifically in the Cy5 channel, dependent on the specific LED light engine and LED configuration:

DA/FI/TR/Cy5-4X-B
Normalized to Lumencor Spectra X
LED-DA/FI/TR/Cy5-4X-A
Lumencor Spectra X
DAPI 100% 134%
FITC 100% 102%
TRITC 100% 135%
Cy5 100% 750%
Signal Normalized to Semrock DA/FI/TR/Cy5-4X-B illuminated with Lumencor Spectra X.

 

DA/FI/TR/Cy5-4X-B
Normalized to CoolLED pE-4000
LED-DA/FI/TR/Cy5-4X-A
CoolLED pE-4000*
DAPI 100% 104%
FITC 100% 166%
TRITC 100% 134%
Cy5 100% 124%
Signal Normalized to Semrock DA/FI/TR/Cy5-4X-B illuminated with CoolLED pE-4000.
*CoolLED pE-4000 configured with 380, 470, 550 & 635 for LED-DA/FI/TR/Cy5-4X-A & with 660 substituted in Cy5 channel for DA/FI/TR/Cy5-4X-B

 

DA/FI/TR/Cy5-4X-B
Normalized to Lumen Dynamics X-Cite® XLED1
LED-DA/FI/TR/Cy5-4X-A
Lumen Dynamics X-Cite® XLED1**
DAPI 100% 99%
FITC 100% 204%
TRITC 100% 132%
Cy5 100% 289%
Signal Normalized to Semrock DA/FI/TR/Cy5-4X-B illuminated with Lumen Dynamics X-Cite® XLED1.
**Lumen Dynamics X-Cite® XLED1 configured with UVX, BDX, GYX & RLX for LED-DA/FI/TR/Cy5-4X-A & with RDX substituted in Cy5 channel for DA/FI/TR/Cy5-4X-B