Bioluminescent Probes for Imaging Biology beyond the Culture Dish

Abstract

Bioluminescence with luciferase-luciferin pairs is an attractive method for surveying cells in live tissues and whole organisms. Recent advances in luciferin chemistry and luciferase engineering are further expanding the scope of the technology. It is now possible to spy on cells in a variety of deep tissues and visualize multicellular interactions—feats that are enabling new questions to be asked and new ideas to be explored. This perspective piece highlights recent successes in bioluminescent probe development and their applications to imaging in live cells, tissues, and animals.

Pyridone Luciferins and Mutant Luciferases for Bioluminescence Imaging

“…d ‐Luciferin is not optimally cell‐permeant, requiring large doses to saturate Fluc and achieve robust photon production in cultured cell and tissue models . Imaging with Fluc has also been historically limited to visualizing one feature at a time, owing to a lack of distinguishable probes . To craft improved bioluminescent tools, many groups have relied on Fluc mutagenesis to isolate brighter and spectrally resolved luciferases .…”

Advances in bioluminescence imaging: new probes from old recipes

“…[4,6] Even fewer have been coopted for use in heterologous systems. [1] Continued efforts to mine new luciferase and luciferin architectures from natural sources are expanding the number of available tools. For example, the luciferase gene from Photinus scintillans was recently cloned.…”

“…[1] Luciferases catalyze light emission via the oxidation of small molecule substrates (luciferins). Since no external light is required, the background emission is virtually zero, enabling sensitive imaging in vivo .…”

In vivo optical imaging of cancer cell function and tumor microenvironment

“…Luciferases derived from firefly and Renilla have been widely used in reporter assays to measure the expression of target genes in cell culture. Cells that stably express luciferase protein as a result of introduction of a luciferase gene produce biopermeable luminescence after giving a substrate such as d ‐luciferin or coelenterazine . With the development of ultra‐sensitive charge‐coupled device (CCD) cameras, it has become possible to detect a trace amount of photons emitted within an animal's body.…”