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.
Lumencor’s manufacturing operations remain healthy and active. While daily operations are inevitably constrained by measures taken to foster good health and minimize the transmission of COVID-19, productivity remains high. Minimal component shortages have developed as shelter-in-place practices have dampened our suppliers’ activities. However, we expect our supply chain will likely suffer additional shortages. Lumencor’s practices now include the quarantine of all receivables as our first commitment is to ensure the safety of our employees while safeguarding their work. In some cases, that may mean that our deliveries to customers will be delayed. We will do our best to inform all customers with pending orders of changes to anticipated shipment dates. Please notify us of any urgency around your order and we will do our very best to assist.
Please be assured that we are watching and managing your business with great care during the current emergency. Please contact to our sales, customer service or technical support representatives if you have any questions or wish to discuss deliverables for your organization’s needs.
A Spectacular Triple Play: LIDA Light Engine®, NIS Elements and sCMOS Cameras for Color Light Microscopy
For histologists, clinical pathologists and anyone seeking improvements in the speed, sensitivity and precision of transmitted light microscopy, Lumencor’s LIDA light engine in combination with Nikon NIS Elements software enable high-speed color imaging data without the need for a dedicated color camera. Instead, monochrome images generated by sequential triggering of the LIDA’s red, green and blue light sources by a sCMOS camera are processed by NIS Elements, delivering video-rate color output. These capabilities allow rapid and fully automated imaging of large tissue sections, as illustrated below. The software also enables convenient switching between camera-synchronized RGB illumination and white-light illumination for ocular viewing. Our application note RGB Color Imaging using the LIDA Light Engine and NIS Elements outlines hardware set-up for Hamamatsu ORCA-Flash4.0 and Andor Zyla sCMOS cameras and provides instructions for image acquisition control using NIS Elements software.
Color image of a 1.5 cm x 1 cm section of adenocarcinoma from human breast acquired using Lumencor’s LIDA light engine and NIS Elements software. Image courtesy of Dr. Michael Weber (Harvard Medical School).
Lumencor’s LIDA light engine mounted on the transillumination port of a Nikon Ti2 microscope with Andor Zyla 5.5 megapixel sCMOS camera
Precise power regulation on each of
Lumencor’s newest and brightest light engines: AURA®, SPECTRA®, CELESTA®, and ZIVA light engines®
In addition to high power and intense brightness, output power regulation is one of the many advanced control features incorporated in Lumencor’s next generation products: SPECTRA, AURA, CELESTA, and ZIVA light engines. To use power regulation, a desired power reference value in milliwatts is entered in the onboard control GUI, as shown in the attached link. To activate power regulation, click the padlock icon next to the reference power value. Gray shading of the padlock icon and the reference power value shows that power regulation is active for the selected output channel. When power regulation is active, the intensity setting for the channel is controlled by the onboard microprocessor, based on feedback from the light engine’s reference photodiode array. The microprocessor continuously adjusts the intensity setting so that the output power matches the power reference value set by the user.
Performance of the output power regulation feedback system of a SPECTRA light engine is illustrated below. The response time of the feedback system is approximately one second. Output power regulation allows users to eliminate variations in light output due to temperature fluctuations and other environmental factors in photometric and quantitative imaging applications where reproducibility and accuracy are essential.
Teal (510/25 nm) channel output from a SPECTRA light engine with and without power regulation. Power output from the light guide was monitored with a Coherent PowerMax II-TO power meter with model PM3 thermopile detector.
In 2018, Lumencor introduced the CELESTA light engine incorporating 7 individually addressable laser light sources. The ZIVA light engine, introduced at the 2019 Society for Neuroscience meeting in Chicago, while similar in some respects, is distinctive in being designed to couple into smaller bore optical fibers.
The output of the ZIVA light engine is suitable for structured illumination microscopy (SIM) and other super-resolution microscopy techniques. In these applications, it provides an alternative to more costly and hard-to-align single mode laser sources. The larger illumination field of the CELESTA light engine is preferred for applications such as spinning disk confocal microscopy, MERFISH or smFISH.
Same onboard microprocessor-based control and feedback interface
Same laser safety interlock configuration
100X widefield (a) and structured illumination microscopy (b) images of actin (green) and mitochondria (orange) in fixed bovine pulmonary endothelial cells. From Pospíšil et al, GigaScience (2018) 8:1–12
This year Lumencor launched the Light Microscopy Imaging Competition in celebration of Earth Day to highlight Lumencor’s commitment to manufacturing bright, clean, and mercury-free light engines. We were impressed with the quality and breadth of applications in the submissions we received. Without further ado here are the winners for Lumencor’s 2019 Earth Day Light Microscopy Imaging Competition:
1st Place – Robert Zucker
EPA, Research Triangle Park, NC
PRIZE: NEW SOLA SE U-nIR Light engine
ARPE 19 cell treated with 75nm silver nanoparticles that are coated with PVP Combination of darkfield image (white nanoparticles) and fluorescence image of nuclei (blue), Golgi (green), and microtubules (red). Observation with a using a 60X plan Fluor lenses and a SOLA light engine®. The image was processed using Nikon deconvolution algorithms.
2nd Place – Glyn Nelson
University of Newcastle, UK
PRIZE: $1000 off voucher
Image of a mosquito larva’s ‘mustache’, the fine brush hairs on the side of their mouth. Z stack deconvolved and rendered in Huygens. Larva fixed in PFA and mounted in glycerol. Imaged with a Leica DMi8 equipped with a 40X oil 1.3 NA objective, GFP filter cube, Hamamatsu Flash4 v2 camera and a SOLA SE light engine.
3rd Place – Abdul Mohammed
Ultivue, Cambridge, MA
PRIZE: $500 off Voucher
16-plex biomarker profiling of Non-Small Cell Lung Cancer tissue (CD3, CD4, CD45RO, CD68, CD163, CD8, CD11c, CD20, PD1, PDL1, Cytokeratin, LAG3, FoxP3, Ki67, GranzymeB and MHCII). Nuclear counterstain shown in blue. The slide was processed using Ultivue’s InSituPlex technology and imaged on a fluorescence microscope using a 20X objective and SPECTRA X light engine®.
These images clearly demonstrate the quality of data attainable by using Lumencor’s solid state light engines. For more information on how Lumencor light engine can help you in you illumination needs please contact us.
This year Lumencor has once again supplied state-of-the-art solid state light engines to support the MBL Physiology course in teaching our next generation of scientists. One light engine showcased on the Nikon Ti2 microscope with the CrEST X-Light V2 spinning disk confocal system was the CELESTA light engine (Figure 1). The CELESTA light engine delivers approximately 1 watt of output power from each of its 7 individually addressable solid-state laser light sources (7 watts total). When coupled to the CrEST X-Light V2 confocal system three-dimensional, multi-colored, image sets such as the one shown below (Figure 2) can be captured by users. To take a look at a video generated from multiple z sections made from the imaging of a late blastula stage of the sea urchin Arbacia punctulata click here.
Figure 1: Nikon Ti2 with CrEST X-Light V2 and CELESTA light Engine
Figure 2: Various Z-sections of a late blastula stage of the sea urchin Arbacia punctulata stained for microtubules (red), actin filaments (green), and DNA (blue).