Confocal microscopy
libst | Louvain-la-Neuve
Picture credit : Yvan Flamant
Confocal microscopy is an optical imaging technique allowing the acquisition of contrasted images of a single plane of a sample.
A laser beam scans the specimen pixel by pixel and line by line and the emitted light is going through a pinhole located in front of the detector to eliminate the light emerging outside that plane.
Only the light within the focal plane can reach the detector. The resulting image has a high contrast and resolution.
This technique allows the acquisition of images in the X, Y and Z planes of a specimen that can be combined into a 3D representation.
The recent improvement of the equipment and the development of increasing array of fluorescent molecules offer the possibility to detect simultaneously several dyes or fluorophores at high sensitivity and to follow their dynamics in living samples.
Equipment
Confocal microscopy
- LEICA STELLARIS 8 FALCON
- White Light Laser 440nm-790nm
- Diodes 405-488-561-638
- AOBS beam splitter
- Thermostatic chamber
- Software LasX STELLARIS and LasX 3D Visualisation
- ZEISS LSM980: Lasers 405-445-488-514-561-663
- Airyscan II/Multiplex
- Definite focus 2
Epifluorescence microscopy
- 2 Inverted Microscopes ZEISS Observer Z1:
- Motorized stage / HXP Lamp
- Bright Field-Phase contrast-DIC-Fluorescence
- Zen software and Zen Module Tiles/Positions
- Microfluidic system CellASIC ONIX 2
Main applications
Confocal microscopy allows the determination of the cellular and subcellular localization of proteins and biomolecules at high resolution, and to follow their dynamics.
In addition, combined with appropriate softwares, this equipment allows to obtain quantitative data on the molecule of interest. Multicolor imaging can be performed with a large array of fluorescent protein.
- Live cell imaging
- Multifluorescence
- Co-localization
- 3D and 4D imaging
- Spectral imaging
- Fluorescence recovery after photobleaching (FRAP)
- Fluorescence lost in photobleaching (FLIP)
- Förster resonance energy transfer (FRET)
