Microscopy Unit

The IBMCC Microscopy Unit provides the followingservices:
1) Confocal microscopy: two confocal microscopes,one Zeiss LSM510 and one brand new Leica SP5.
2) Live cell imaging and Deconvolution: one Olympus IX71 with Delta-Vision system, for live cell imaging. It carries a chamber that controls CO2 and temperature.
3) Live cell imaging: one Nikon Eclipse TE-2000 fluorescent microscope with a chamber with CO2 an temperature control too.
4) The Unit also looks after additional microscopes in the IBMCC: 9 fluorescent microscopes, 10 inverted microscopes for tissue culture, 1 microinjector, 1 microscope for cytogenetic and 1 microscope for hystology.

The Unit personnel:
• Runs the confocal microscopes doing Z-Series, Timelapse, FRAP y FRET.
• Gives tutorials on the correct use of the microscopes and deals with the day to day problems.
• Helps the users with the different imaging software: MetaMorph®, Openlab®, AxioVision®, LSM Image Browser® y DPManager®.
• Carries out monthly revisions of the different microscopes, including phase adjustment, Kohler adjustment and objective cleaning and weekly revisions of the different microscope rooms (material reposition).
• Changes and fixing of the mercurium and halogen lamps.
• Writes guides and tutorials on the use, basic configurations and programmes on the Unit website.
• Looks after the Unit quality system. ISO Certifications:ISO 9001:2000.

• Confocal microscopy to external users. This service offers microscopy both to internal and external users. Electronic management allows remote communication
with the service to book the use of different microscopes, especially confocals.
• Training and advice on the use of programs for image capture. Training and advice on the use of image capture programs, including basic training and solving of inquiries about the programs of capture, image analysis and processing including: Metamorph,Leica LAS AF,Image Browser LSM and Openlab.
• Creating, updating and maintaining the Web Unit. Creating, updating and maintaining the Web unit, including a constant update of technical specifications for new equipment that will incorporate the center, as well as incorporation of tutorials and guides. Basic use of major hardware and software associated analysis. Image for downloading by users of the center.
• Advice and training in the use of equipment for living cell microscopy (Nikon Eclipse TE2000). Advice and training in the use of equipment for living cell microscopy (Nikon Eclipse TE2000) with a system of CO2 and temperature control, including basic training in the handling of the team, as well as use and associated software configuration (Metamorph), assembling and designing experiments Timelapse and reconstruction of processing images and videos end.
• Development of Tutorials. Development of Tutorials with instructions for the use of image analysis programs to facilitate the use of microscopes and the capture and image analysis in associated programs.
• Review and monthly maintenance of equipments of microscopy. Review and monthly maintenance of equipment of the Center for microscopy, including setting stage for microscopy in cell culture rooms and adjustment for Kohler for fluorescence microscopes. Control, replacement and adjustment of fluorescent lamps of fluorescence microscopes.

The Microscopy Unit of Cancer Research Center owns a great collection of microscopes that offers a high range of analysis possibilities. There are four main equipments due to their cost and the service they give:
• Laser Scan Confocal Microscopy Leica SP5. The machine was co-paid by FEDER, Ministerio de Sanidad y Consumo and Instituto de Salud Carlos III. It is the Unit main equipment – nearly 80% of the staff job is related to it. The microscopy has four lasers which have seven excitation lines, which, in combination with the spectral detection technology, allows the observation of every fluorocrome in the visible range. Due to its confocal module, it is the most demanded facility by users in order to obtain high resolution images of cell cultures or tissues. Besides, due to its software and the motorized stage techniques such as FRAP, FRET or co-localization analysis can be performed.
• Live Cell Microscopy Delta-Vision. Acquired with FEDER and CSIC founding, this machine is mainly used in in vivo timelapse experiments. Its principal application is observation and image acquisition of yeasts (cell-wall eukaryotic organisms).
Anyway, this equipment is widely used-experiments in animal cells, cultures or fixed cells are run as well. The microscopy main advantages are the ultra-precise stage, the lighting system which combines a xenon lamp with a excitation filters wheel, and a high sensitive CCD camera. Hence, the machine acquires images with high speed and low exposition time, which decreases the damages that the live cells can suffer. In the other hand, the deconvolution module treatment gets high quality images.
• Laser Scan Confocal Microscopy Zeiss LSM 510. It’s older equipment and the demand for its use has decreased since the arrival of the Leica SP5. It’s useful when the demand is over possibilities of the Leica and for external users, who can use
this microscopy under technician unit control.
• In vivo experiments microscope Nikon TE2000. This equipement is very useful to performance asays with living cells. In addition, this microscope is combined with Metamorph, a very powerfull tool in microscopy. This software provides a lot aplications to transfor images, making measurements and many things usisng microscopy images.In addition, the Microscopy Facility of the Cancer Research Center is completed with much equipment of fluorescence, inverted microscopes for culture checking and microscopes for FISH and histological analysis.

Scientific Coordinator
Atanasio Pandiella
Tel.: 923 294 815
E-mail: atanasio@usal.es
Sergio Moreno
Tel.: 923 294 810
E-mail: smo@usal.es

Location and contact
Unidad de Microscopía