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Stories & Features

Semrock Introduces the BrightLine® Family of LED-based Light Engine Filter Sets

Aug 6, 2014 by IDEX Health & Science

Four ExcitersLED-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 fourteen sets in single-band, full Pinkel & Sedat 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). This state-of-the-art set represents a break-through in quad-band performance, and has been optimized for popular LED-based light engines from manufacturers including Excelitas Technologies®, CoolLED, and lumencor®. 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 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 are  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.


Comparison of excitation and emission wavelengths, and dichroic edge positions by part number for a traditional light source set versus the LED-based light engine optimized filter set:

 DA/FI/TR/Cy5-4X-BLED-DA/FI/TR/Cy5-4X-A
DAPI-ExciterFF01-387/11FF01-392/23
FITC-ExciterFF02-485/20FF01-474/27
TRITC-ExciterFF01-560/25FF01-554/23
Cy5-ExciterFF01-650/13FF01-635/18
DichroicFF410/504/582/669-Di01FF409/493/573/652-Di01
EmitterFF01-440/521/607/700FF01-432/515/595/730

 

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
DAPI100%134%
FITC100%102%
TRITC100%135%
Cy5100%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*
DAPI100%101%
FITC100%206%
TRITC100%131%
Cy5100%283%
Signal Normalized to Semrock DA/FI/TR/Cy5-4X-B illuminated with CoolLED pE-4000.
*CoolLED pE-4000 configured with 385, 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**
DAPI100%99%
FITC100%204%
TRITC100%132%
Cy5100%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