Equipment
GLiM offers accessibility to a divers set of microscopes. They are subdivided below by the type of microscopy.
Reservation of the instruments occurs through CFMS.
Manuals, SOPs and background information can be accessed on the sharepoint site after an access request.
Wide-field microscopes
Wide-field microscopy is used for thin samples, provides low light exposure and can acquire frames with a speed in the order of 20fps. With these microscopes, the sample is illuminated by a collimated beam of light.
Confocal microscopes
Confocal microscopy is used to provide optical sectioning and an improved resolution. The sample is illuminated by a focused laser beam which is scanned over the sample. Emission from each position is detected by a point detector and a pinhole removes out of focus emission light to improve contrast and resolution.Spinning disk microscope
Spinning disk microscopes can acquire images with a speed in the order of 20fps and a reduced light dose compared to confocal microscopes while providing optical sectioning. While confocal microscopy uses one pinhole to detect fluorescence of a sample scanned by one excitation spot, spinning disk confocal microscopy uses a rotating disk with 1000 or more pinholes to generate multiple simultaneous excitation beams. Thereafter emission is detected through the same set of pinholes to ensure confocal detection.
Multi-photon microscope
Multi-photon microscopy is advantageous for deep imaging in thick (living) samples and is an alternative to confocal microscopy. Fluorophores are excited with light of a long wavelength (half the energy needed for one photon excitation) and the fluorophore needs to absorb multiple photons simultaneously. These events can only occur at high excitation intensities which can be reached in the focal volume of a pulsed laser, thereby eliminating out-of-focus excitation and fluorescence. Two photon microscopy is advantageous for optical sectioning in thick samples.
Laser capture microdissection
Instruments which allow isolation of individual (living) cells, a piece of tissue or even subcellular components from a sample under light or fluorescence microscopic visualization. This isolation occurs without direct contact minimizing risk of contamination.
Nanoscopy
Since the invention of the first microscopes almost 400 years ago, there has been an ongoing strive to ever see more details within organisms, tissues and cells. Based on fundamental physics from the nineteenth century, it was long thought impossible to see beyond a resolution of ~200 nm with light microscopes. However, thanks to recent advances in fluorescent labels and laser technology, it is now possible to improve this resolution by a staggering factor of 10, making ‘nanoscopy’ a reality.
Custom-made microscopes
Microscopes which are not commercially available and designed for a specific research purpose.