If there is one thing which always sparks our interest at GoldBio, it’s the novel methods and discoveries that proliferate in our biotechnology field! Take luciferase for example. Though this compound was detailed half a century ago and has been in increasing use in bioluminescent assays for decades, scientists are still discovering new ways to improve its viability and usefulness in the lab. I recently came across two papers, both looking for a way to look at two different bioluminescent markers on the same cell; both searching in the same direction but from completely different points of view.
The first, published in PLoS One in 2011, is from Laura Mezzanotte, et al.(1). In it, they discussed the use of dual color bioluminescence by way of a newer, red codon optimized, firefly luciferase (Ppy RE8) along with the traditional green luciferase (CBG99)! The red luciferase has a distinct emission peak at 618nm compared to the CBG99 emission peak at 537nm, which is a good separation for spectral analysis in systems that normally have low quantum efficiencies.
Also, the red bioluminescence was not absorbed as easily by vascularized tissue, traditionally a problem with the blue light from the Renilla and Guassia luciferases. So their luciferase-encoded cells performed wonderfully in vivo when they were injected into lab mice. This dual-color luciferase system gave the researchers the tremendous advantage of a simplified method which required only one substrate, D-luciferin, to fuel both genes (which YOU can find the best deal conveniently at Gold Biotechnology (Catalog# LUCK). What could be better than that?
The second paper, publish in Biotechniques in 2010, delved more into the engineering side of multiple gene expression. HyuckJoon Kwan, et al.(2) were more interested in modifying their CCD camera into a dual-path luminescence imaging system. With it, they could detect the emission spectrum of green- and red-emitting luciferase in two different genes at the same time! Their system used a series of dichroic mirrors to divide the incoming green and red light wavelengths. Because of the split, the emissions were focused on different areas of the CCD by the imagining lens. That meant the single CCD could obtain two images of the cell simultaneously, one in red and the other in green!
Wouldn’t it be amazing if you could see the precise interaction of enzymes in two cells at the exact same time instead of taking a green filter pass and then a red filter pass with your camera and assuming (hoping) what was happening while you were filming in the other filter? And just think if Mezzanotte and Kwan had come together to use both of these novel systems in a dual purpose! As always in science…the possibilities are endless.
- Mezzanotte, Laura, et al. "Sensitive dual color in vivo bioluminescence imaging using a new red codon optimized firefly luciferase and a green click beetle luciferase." PLoS One 6.4 (2011): e19277.
- Kwon, HyuckJoon, et al. " Bioluminescence imaging of dual gene expression at the single-cell level." Biotechniques 48 (2010): 460-462.
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