Related Experiment Video
Updated: May 6, 2026
07:33
In vivo Dual Substrate Bioluminescent Imaging
Published on: October 11, 2011
15.9K
Multicomponent Bioluminescence Imaging with a π-Extended Luciferin.
Journal of the American Chemical Society
|August 14, 2020
Summary
Researchers developed new bioluminescent probes using engineered luciferases and π-extended luciferins. This breakthrough enables multicolor imaging and deep-tissue visualization, overcoming limitations of traditional bioluminescence imaging (BLI).
Area of Science:
- Biochemistry
- Molecular Biology
- Biotechnology
Background:
- Bioluminescence imaging (BLI) using luciferase-luciferin systems is vital for studying biological processes.
- Existing BLI probes face limitations in distinguishing signals and deep-tissue penetration.
- Need for novel probes to enable multicolor and deep-tissue imaging.
Purpose of the Study:
- To engineer novel, red-shifted bioluminescent probes for enhanced imaging capabilities.
- To develop orthogonal probes for simultaneous multicolor imaging.
- To utilize Rosetta-guided enzyme design for creating complementary luciferase-luciferin pairs.
Main Methods:
- Synthesis of π-extended luciferins designed as rotationally labile luminophores.
- Identification of a luciferin analog with an intramolecular 'lock' mechanism.
- Rosetta-guided enzyme design to create a complementary luciferase for the novel luciferin.
Main Results:
- A novel, red-shifted luciferase-luciferin pair was successfully engineered.
- The new probe exhibits orthogonality, allowing for multicomponent imaging.
- Demonstrated simultaneous imaging of four substrate-resolved luciferases in one session.
Conclusions:
- This study presents the first use of Rosetta-guided design for engineering bioluminescent tools.
- The developed probes significantly expand the scope of orthogonal bioluminescence imaging.
- The engineered system overcomes limitations of traditional BLI, enabling advanced biological visualization.
Related Concept Videos
01:18Imaging Biological Samples with Optical Microscopy
9.2K
Optical microscopy uses optic principles to provide detailed images of samples. Antonie van Leeuwenhoek designed the first compound optical microscope in the 17th century to visualize blood cells, bacteria, and yeast cells. In 1830, Joseph Jackson Lister created an essentially modern light microscope. The 20th century saw the development of microscopes with enhanced magnification and resolution.
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...
In optical microscopy, the specimen to be viewed is placed on a glass slide and clipped on the stage...
9.2K
01:16Confocal Fluorescence Microscopy
16.0K
Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
16.0K
01:05Total Internal Reflection Fluorescence Microscopy
11.0K
Total internal reflection fluorescence microscopy or TIRF is an advanced microscopic technique used to visualize fluorophores in samples close to a solid surface with a higher refractive index, such as a glass coverslip. TIRF only allows fluorophores in proximity to the solid surface to be excited. When light from a medium with a lower refractive index (such as air) hits the glass coverslip at a critical angle, the light undergoes total internal reflection stead of passing through the glass.
11.0K
01:37Super-resolution Fluorescence Microscopy
12.3K
Super-resolution fluorescence microscopy (SRFM) provides a better resolution than conventional fluorescence microscopy by reducing the point spread function (PSF). PSF is the light intensity distribution from a point that causes it to appear blurred. Due to PSF, each fluorescing point appears bigger than its actual size, and it is the PSF interference of nearby fluorophores that causes the blurred image. Various approaches to achieving higher resolution through SRFM have recently been...
12.3K