Multi-modal advanced light microscopy

Deconvolution widefield microscopy

Description
Deconvolution is a computational technique for improving the contrast and resolution of digital images. It includes a suite of methods that seek to remove or reverse the blurring present in microscopes images caused by the limited aperture of the microscope objective lens. Nearly any image acquired on a digital fluorescence microscope can be deconvolved.
In addition, new applications to transmitted light images are now available. Three-dimensional images made up of a series of optical sections are particularly well suited for improvement by deconvolution.

Service provider
Advanced Light Microscopy Facility (ALMF)

Location
Heidelberg, Germany

Contacts
Scientific contact: Rainer Pepperkok
Technical contact: Stefan Terjung

Laser scanning confocal microscopy (LSCM / CLSM)

Description
Laser scanning confocal microscopy (LSCM), also known as Confocal laser scanning microscopy (CLSM) or simply confocal microscopy, is a technique for obtaining high resolution optical images with depth selectivity. The key feature of confocal microscopy is its ability to acquire in-focus images from selected depths, a process known as optical sectioning. Images are acquired point-by-point and reconstructed with a computer. This allows three-dimensional reconstructions of topologically complex objects.

Service provider I
Advanced Light Microscopy Facility (ALMF)

Location
Heidelberg, Germany

Contacts
Scientific contact: Rainer Pepperkok
Technical contact: Stefan Terjung

 

Service provider II
The Institute of Photonic Sciences (ICFO)

Location
Barcelona, Spain

Contacts
Scientific Contact: Pablo Loza-Alvarez 
Technical Contact: Jordi Andilla, Maria Marsal 

Multiphoton microscopy systems

Description
Multiphoton microscopy is a fluorescence imaging technique that allows imaging of living tissue up to about one millimeter in depth. It uses red-shifted excitation light, which can also excite fluorescent dyes. However, for each excitation, two photons of low energy infrared light are absorbed simultaneously to provide the required energy for electrons in the fluorophore to reach the excited state. Using infrared light minimizes scattering in the tissue. Due to the multiphoton absorption, the background signal is strongly suppressed. Both effects lead to an increased penetration depth for these microscopes.

Service provider I
Advanced Light Microscopy Facility (ALMF)

Location
Heidelberg, Germany

Contacts
Scientific contact: Rainer Pepperkok
Technical contact: Stefan Terjung

 

Service provider II
The Institute of Photonic Sciences (ICFO)

Location
Barcelona, Spain

Contacts
Scientific Contact: Pablo Loza-Alvarez
Technical Contact: Jordi Andilla, Maria Marsal

Spinning disc confocal microscopy (SDCM)

Description
Spinning disc microscopy has advanced significantly in the past decade and now represents one of the optimum solutions for both routine and high-performance live-cell imaging applications. Spinning disc confocal microscopy technique uses a series of moving pinholes on a disc to scan spots of light. Since a series of pinholes scans an area in parallel, each pinhole is allowed to hover over a specific area for a longer amount of time thereby reducing the excitation energy needed to illuminate a sample when compared to laser scanning microscopes. Decreased excitation energy reduces photo-toxicity and photo-bleaching of a sample often making it the preferred system for imaging live cells or organisms.

Service provider I
Advanced Light Microscopy Facility (ALMF)

Location
Heidelberg, Germany

Contacts
Scientific contact: Rainer Pepperkok
Technical contact: Stefan Terjung

Total internal reflection fluorescence microscopy (TIRF)

Description
Total internal reflection fluorescence microscopy (TIRF) is a type of microscopy technique with which a thin region of a specimen, usually less than 200 nm can be observed. A TIRF microscope uses an evanescent wave to selectively illuminate and excite fluorophores in a restricted region of the specimen immediately adjacent to the glass-water interface. The evanescent wave is generated only when the incident light is totally internally reflected at the glass-water interface. The evanescent electromagnetic field decays exponentially from the interface, and thus penetrates to a depth of only approximately 100 nm into the sample medium. Thus the TIRF microscope enables a selective visualization of surface regions such as the basal plasma membrane (which are about 7.5 nm thick) of cells.

Service provider I
Advanced Light Microscopy Facility (ALMF)

Location
Heidelberg, Germany

Contacts
Scientific contact: Rainer Pepperkok
Technical contact: Stefan Terjung

 

Service provider II
The Institute of Photonic Sciences (ICFO)

Location
Barcelona, Spain

Contacts
Scientific Contact: Pablo Loza-Alvarez
Technical Contact: Jordi Andilla, Maria Marsal