Dichroic Mirrors and Filters for SPECTRA, CELESTA, and ZIVA Light Engines®
In fluorescence microscopy, the filter set, consisting of excitation and emission bandpass filters and a dichroic beamsplitter, plays a critical role. It performs the essential functions of directing excitation light from the light source to the sample and then separating it on the basis of wavelength, from fluorescence emitted from the specimen. The filter set consists of excitation and emission bandpass filters and a dichroic beamsplitter. Optimized filter sets are critical because fluorescence emission from a microscopic specimen ismany orders of magnitude (>106) weaker than the excitation from the light source. Recognizing optimized filter set specifications are critical for obtaining images with high signal:background contrast. Lumencor has developed uniquely high performance, multiband dichroic beamsplitters and multiband and single band emission filters for use with our SPECTRA, CELESTA, and ZIVA light engines®. These light engines are setting the standard for high performance, high brightness, turn-key solutions in solid-state lighting for life and industrial sciences.
All emission filters and dichroics have standard dimensions and are supplied in an unmounted form. Installation in filter cubes, filter wheels, or other instrumentation-specific mountings is required before use.
The multiband emitters and dichroics are optimized for compatibility with the electronically selectable excitation outputs of SPECTRA, CELESTA, and ZIVA light engines® (Figure 1). This enables fast multicolor imaging without the need for filter wheels or other positioning devices to execute filter interchanges. Single bandpass filters are offered for use in situations where fluorescence crosstalk (e.g. detection of FITC fluorescence derived from violet (DAPI) excitation) confounds identification of fluorescently labeled components of the specimen.
Figure 1. Transmission spectra of CELESTA/ZIVA VCGRnIR pentaband dichroic and emitter superimposed on the violet, cyan, green, red, and near-infrared output lines of the CELESTA and ZIVA light engines®.
Lumencor’s Earth Day Light Microscopy Imaging Competition is in full swing, offering the opportunity to win up to $10,000 worth of state-of-the-art, solid-state lighting! Submit your images today and help us celebrate and promote a brighter, greener planet through the use of mercury-free illumination. Qualifying images must be acquired using Lumencor light engines.
Lumencor can support your laboratory as you return to work. The challenges are numerous for researchers and instrument manufacturers alike- maintain best lab practices, manage constrained budgets, keep staff safe and productive. Let Lumencor help with illumination tools designed for the best data quality and best lab safety!
Multiplexed imaging of gene expression using the CELESTA light engine
MERFISH (multiplex error robust fluorescence in situ hybridization) is an imaging technique that profiles cell populations based on the identification of thousands of RNA transcripts per cell.The CELESTA light engine is an ideal and widely-adopted illumination source for this application.In a recent paper published in Nature , Wheeler and co-workers used MERFISH imaging with a CELESTA light engine to quantify the expression of nine specific astrocyte and T-cell markers.Five of the CELESTA light engine’s seven laser lines were used in the highly multiplexed MERFISH imaging protocol.The overall objective of the research described in the paper was to characterize astrocyte populations that contribute to pathogenesis in a preclinical model of multiple sclerosis.
 MA Wheeler, JR Moffitt, IC Clark, EC Tjon, Z Li, SE J Zandee, CP Couturier, BR Watson, G Scalisi, S Alkwai, V Rothhammer, A Rotem, JA Heyman, S Thaploo, LM Sanmarco, J Ragoussis, DA Weitz, K Petrecca, JR Moffitt, B Becher, JP Antel, A Prat, FJ Quintana, Nature (2020) 578:593–5990
Independent Intensity and Pulse Width Control for Stroboscopic Illumination
Evaluation of photo-stimulation intensity dependence is often a necessary part of neuromodulation experimentation utilized in optogenetics studies . The inherent stability and quantitative nature of Lumencor’s SPECTRA X light engine make it particularly well suited as the pulsed light source of choice for studies requiring pulse width and frequency of stroboscopic illumination analyses. Find more detail regarding this extremely stable, reproducible, and well-behaved data, as well as a specific reference in a recent Journal of Physiology publication by authors Kubota, Sidikejiang, and Seki, on Lumencor’s website.
Figure Description: Alternating cyan (485/25 nm, ~0.5 ms) and green (560/32 nm, ~3 ms) output pulses generated by TTL triggering of a SPECTRA X light engine. Two superimposed oscilloscope traces are shown in which the cyan intensity is adjusted from 100% to 55% via RS232 serial commands while the green intensity remains constant. Separation of the cyan and green pulses is ~0.25 ms.