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Research Technical Services

Confocal Microscopy

Technique of microscopic observation

Confocal Laser Microscopy is a microscopic observation technique that is based on obtaining images of the emission of fluorescent light from the sample, from which the light from the out-of-focus planes is removed, collecting only the focal plane light. This has great advantages over conventional microscopy, such as greater contrast, sharpness and resolution of the images, and the possibility of obtaining optical sections of the sample to perform three-dimensional studies.

Job request

In order to request that a sample be analysed, you need to contact the staff responsible for the team.

You will need to give the toxicity and any storage precautions. Once the report has been delivered, if you do not collect the sample within one week, any remaining sample will be destroyed.

The equipment

NIKON A1R

Year of acquisition:
2013
Spectral Confocal Laser Microscopy (CLSM) module installed in a high-performance motorised inverted microscope equipped with:

- Bank of lasers

- Scanning module

- Motorized turntable

- Control modules for photomultiplier and spectral detectors

- Cell incubation system

- Control program NIKON NIS-Elements AR v 4.10


Approximate resolution:
150 nms in XY and 250 nms in Z

It is a spectral confocal laser microscopy (CLSM) module and a structured light microscopy module of the "Spinning disc” type. Both modules have been installed on a high performance motorised inverted microscope.

How does it work?

Technical characteristics of the instrumental set:

  1. NIKON Ti Eclipse motorised inverted microscope, with 6 Plan Apochromatic objectives (10x, 20x, 40x, 60x water, 60x oil, 100x oil), phase contrast and DIC. Piezoelectric plate with up to 1nm resolution in the Z axis.
  2. Lighting systems: halogen white transmitted light source, fiber optic epifluorescence and turret for six filter blocks, argon laser with 4 emission lines (457, 477, 488 and 514nm), laser Helium-Neon (543nm), UV diode laser (405nm) and red diode laser (642nm).
  3. NIKON A1R confocal module with galvanometric and resonant tracking, of different directionality. Maximum resolution of 4096 x 4096 pixels and maximum speed of 420 fps at 512 x 32 pixels.
  4. Reflected light detectors: 4 photomultipliers and 32-channel spectral module.
  5. DSD ANDOR disc spinning module with structured lighting and high definition CCD camera.
  6. Overplatinum cell incubation system, with CO2 atmosphere, equipped with temperature and humidity control.
  7. Computer HP Z820 for the complete control of the whole instrument and of capture of images Software owner NIS Elements v 4.10
  8. Independent computer to be able to work "off line", intended for image analysis by users.

Applications / Practical cases

Possibility to carry out different fluorescence assays in live cell: FRAP, FRET, PA, Time lapse experiments, colocalization studies,..., and also by means of reflection, observing the structure of materials, pores and microfisures, roughness, three-dimensional fibre studes, and so on.

web images

More information:

All those who are interested in receiving more specific information about the equipment and its applications, can contact the equipment managers.

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