Application SpotLIGHT – January 2021: Time-lapse Imaging with SOLA Light Engine®

The Relevance of Time-lapse Imaging of GFP Expression Using the SOLA Light Engine® to COVID-19 vaccine Efficacy

The delivery of mRNA through lipid-based transfection has been a longstanding challenge for the development of RNA therapeutics. Moreover, it has acquired a new and urgent prominence from the development of COVID-19 vaccines consisting of mRNAs encapsulated in lipid nanoparticles by Pfizer/BioNTech and Moderna. It is clearly important to understand the effects of mRNA-lipid complex formulation and extracellular medium composition on downstream expression of the protein immunogen that in turn determines vaccine efficacy.  In 2019, before the start of the COVID-19 pandemic, a team of researchers from Ludwig-Maximilians-University in Munich and Stony Brook University, New York described the use of live-cell imaging on single-cell arrays (LISCA) to monitor the onset and rate of GFP expression following mRNA lipoplex transfection [1].  Single cells are arrayed on a micropatterned fibronectin substrate (Figure 1A), incubated with mRNA-lipid complexes for 1 hour and then monitored by time-lapse fluorescence microscopy for 20 hours (Figure 1B).  For GFP fluorescence to give an authentic representation of protein expression levels, stable and reproducible excitation is essential, making the SOLA Light Engine the ideal illumination source for this application.  As well as characterizing the pronounced cell-to-cell variability in onset times and rates of protein expression (Figure 1), LISCA was used to determine the effect of serum proteins on the cellular uptake of different mRNA-lipid complex formulations.

Figure 1. (A) Single GFP-expressing HuH7 cells arrayed on micropatterned fibronectin. (B) Single cell fluorescence trajectories representing GFP expression. The gray-shaded area represents the initial 1-hour period of incubation with mRNA-lipid complexes. (C) Enlarged region of (B) showing cell-to-cell variation in onset of protein expression. Reproduced from Reiser et al. (2019) [1] under the terms of the Creative Commons Attribution License.

Reference

[1] A Reiser, D Woschée, JO Rädler et al.  Integr Biol (2019) 11:362–371


Download the PDF of Lumencor Application SpotLIGHT: January 2021

Parallel Light Engine Performance monitoring Using the Onboard Control GUI

Parallel Light Engine Performance monitoring Using the Onboard Control GUI

AURA, CELESTA, RETRA, SPECTRA, and ZIVA Light Engines incorporate a control GUI accessed through a web browser via an ethernet connection.  Image acquisition applications used to control the light engine though connection of either the USB or RS232 serial ports can be run in parallel with ethernet-connected control GUI to aid in trouble-shooting.  As shown in Figure 1, this allows the light engine to be controlled by the image acquisition software, while the GUI serves as a passive monitor of the light engine status.

Figure 1.  Screenshots of parallel operation of image acquisition software (NIS Elements, left) and CELESTA Light Engine Control GUI (right).  A. NIS Elements command to turn on red light output at 21.7% is inoperative.  Examination of the GUI display reveals that this is due to an open interlock condition (e.g. no optical fiber connected to the light engine output port).  B. After closing the interlock, the same NIS Elements command results in light output, indicated by the filled red channel radio button and non-zero output power reading in the GUI.

The control GUI displays many types of information pertinent to the performance of the light engine that are not accessible in current releases of most image acquisition software packages.   These include:

  • Real-time light output power readouts
  • Standby mode status
  • Light engine operating software error messages
  • Humidity/dew point data
  • Serial port configuration
  • TTL port configuration
  • Cumulative run time data

In cases where the PC being used for image acquisition control has a single ethernet port that is dedicated to internet access, a USB-to-ethernet adapter (Figure 2) can be used for connection to the light engine control GUI.  USB-to-ethernet adapters are readily available from online vendors for less than $20.

Figure 2. USB-to-ethernet adapter

 

 

 

 

 

Lumencor, Inc., Named Makers & Manufacturers Honoree in Portland Business journal Product Innovation Awards, 2020

Lumencor, Inc. has been named 2020 Product Innovation of the Year honoree by the Portland Business Journal for our next generation SOLA light engine®. Each year, the PBJ honors the region’s top manufacturing companies who drive the economy with innovation, excellence and productivity. The new generation SOLA features increased power, longevity, stability and robustness over the projected 15 year life time with no replacement parts. Lumencor’s SOLA light engine is used in fluorescence microscopy for life science and materials science applications… Read Press Release

Find out more about the Innovation Award

 

Recommended Operating Conditions for CELESTA, SPECTRA and ZIVA Light Engines®

Recommended Operating Conditions for CELESTA, SPECTRA and ZIVA Light Engines®

 

To support the long-term stability of the laser light sources in CELESTA, SPECTRA and ZIVA light engines it is recommended that they should be operated only in environments where the dew point is below 15ºC. For reference, at a typical room temperature of 24ºC, a dew point of 15ºC corresponds to 57% humidity. The current dew point inside the light engine, calculated from onboard temperature and humidity sensors, is displayed on the settings page of the onboard control GUI.

Standby Mode, previously described in the October 2019 issue of Light Reading, is another control system designed to support the long-term stability of the laser light sources. Consequently, users writing their own light engine control software are strongly advised to NOT programmatically disable standby mode.

 

 

If operational situations arise where it is necessary to avoid the onset of standby mode during a data acquisition process, please contact techsupport@lumencor.com. We will be happy to work with you and our software engineering team to devise appropriate solutions.

 

Download the PDF of Lumencor Light Reading: July 2020

Earth Day Imaging Contest Extended

Every Day is Earth Day at Lumencor!

Here’s your last chance in 2020 to win free mercury-free lighting for a BRIGHTER, GREENER, PLANET

Participate in our Earth Day Light Microscopy Imaging Competition by showing us your best photomicrograph, using a Lumencor light engine as your light source, and you could win up to $10,000 in Lumencor lighting products.

Submission deadline Extended: July 31st, 2020

Back to the Lab

We’re in this COVID-19 fight together.

 

Whether with

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!

  • Only Lumencor offers a 15-year performance projection that eliminates toxic mercury and metal halide bulbs with best-in-class solid-state lamps.
  • Only Lumencor offers uniquely stable outputs to maximize S/N, lower detection limits, and enhance sensitivity as your image and screen.

Lumencor’s solid-state lighting is tailored for the instrumentation employed by those engaged in our fight against COVID-19.

We understand the critical role our products play. We remain committed to supplying our customers as safely, efficiently, and expeditiously as possible.

lumencor
14940 NW Greenbrier Parkway | Beaverton, OR 97006 USA | 503.213.4269 | info@lumencor.com