Confocal microscopy

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

Zeiss LSM 710 confocal microscopy fitted with 7 laser lines (405, 440, 458, 488, 514, 543 and 633 nm) and a scanning module including 34 spectral detection channels
Zeiss LSM Exciter confocal microscope fitted with an argon multilign laser
Workspace station equipped with Zen Software as a user interface for image acquisition
A second workspace station with Zen Software is available for image analysis and quantification

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)