Olympus America has introduced a microscope designed to acquire very low-light bioluminescence images.
The LV200 stand-alone microscope system claims to provide sub-second imaging with exceptional clarity.
Bio- and chemi-luminescence, natural phenomena in many living species, have been used in the life sciences for several decades.
In some ways, luminescence resembles fluorescence, which is among the most widely used imaging methods for research microscopy.
But unlike fluorescence, in which cells are labeled with probes that absorb excitation light at one wavelength and then emit light at another, luminescence relies on a chemical reaction occurring within the specimen.
Bioluminescence occurs in a living cell when an enzyme (luciferase) reacts with a substrate (luciferin or another enzyme-specific substrate) to generate light.
Imaging cells by exploiting this reaction has several advantages over fluorescence.
First, repeated exposure of living cells or tissues to fluorescence excitation wavelengths can harm or kill cells, while bioluminescence is not burdened by such phototoxic effects.
Also, unlike fluorescence, bioluminescence imaging typically does not suffer from unwanted background signal. This is a major advantage when imaging samples with high native background fluorescence such as plants and some mammalian tissues.
The resulting high signal-to-background ratio allows more straightforward signal quantification and more sensitive signal detection. Finally, the excitation light of fluorescence causes photobleaching of known fluorophores. Because no excitation light is required for luminescence imaging, photobleaching is not an issue.
Despite these advantages, applications for luminescence in live cell imaging have been limited until now, primarily because luminescence is much dimmer than fluorescence. Even at its best, light emitted by luciferases is dim compared to light emitted by fluorescent probes.
Luminescence has been used in a variety of ways, including bioluminescence resonance energy transfer (BRET) assays, in which the signal from thousands (or millions) ofcells is collected and quantified.
But until now, imaging single cells has been challenging; it demands a completely dark environment and requires relatively long exposure times.
Olympus designed the LV200 microscope with special optical elements and the shortest-possible light path from sample to detector to maximise light capture and minimise the loss of valuable luminescence photons from living cells. In addition, the space-efficient, benchtop microscope features a built-in live-cell incubator and a light-tight casing that prevents ambientlight from interfering with imaging.
Together, these design elements allow researchers to acquire much brighter luminescence images using much shorter exposure times, even over extended periods. The LV200 has been used successfully to image biological processes in chronobiology, neuroscience, developmental and plant biology, and more.
Now, together with brighter species of luciferases, such as Promega Corporation’s recently launched NanoLuc luciferase, the LV200 makes it possible to use bioluminescence for a far wider range ofexperiments than was ever possible before.
“Researchers have long waited for luminescence imaging to come of age,” said Dr. Michael Lewis, Director of Marketing for Olympus America’s Scientific Equipment Group. “The technique allows high-resolution imaging with unprecedented sensitivity and clarity over long periods. Now, for the first time, both the instrumentation and consumables are available to move science forward using this technique.”
