Equipment

Spinning disk confocal microscopes

The Yokogawa Spinning Disk microscopes allow multicolor fluorescence image acquisition at high speed with low light, by using sensitive Electron-Multiplier-CCD cameras. The systems provide ideal imaging conditions for living cells or weak fluorescent signals.

To facilitate the work-flow both systems operate under Metamorph software on a Nikon inverted microscope equipped with auto-focus and automated stage with multipoint memory and piezo-controlled xyz submicron accuracy. Fast dynamics of fluorescent molecules in cells or samples can be monitored for extended time periods and in 3-dimensions. A FRAP-unit allows local subcellular bleaching to study molecular dynamics. A heating/CO2 unit on stage allows to keep ideal conditions for living animal or human cells.

The specifications of the hardware of the Roper and Andor spinning disk systems are listed in the Technical Details paragraph.

Technical details

Comparison of main specs of Roper and Andor SD systems
Part Andor-Revolution SD Roper SD
Install date 2013 2009
inverted microscope Nikon Ti Eclipse PFS3 Nikon Ti Eclipse PFS2
diode 405 nm CW 100 mW --
DPSS 488 nm CW 50 mW 50 mW
DPSS 561 nm CW 50 mW 50 mW
diode 640 nm CW 100 mW (640 nm) 100 mW (633 nm)
EM-CCD camera Andor iXon888, 1024x1024, 13x13 Photometrics Evolve 512x512, 16x16
xyz stage piezo ASI XY- LE, z- 150um piezo ASI
objectives 10-20-60-100 10-20-60-100
emission filters on request on request
shutter Rotr shutter wheel 30 ms Yokogawa shutter
transmitted light CoolWhite LED halogen
UV Intensilight c-HGFIE Hg
FRAPPA -- YES
5% CO2 and 37C YES, Tokay Hit --

Information obtained by light microscopy

Localisation. Specific staining or fluorescent labels allow temporal and spatial localisation of your (bio)molecule of interest. A range of fluorescent probes can be combined for simultaneous multicolor, multidimensional image acquisition.

Morphology and 3D reconstruction. The shape, 3D volume and interaction of molecules, organelles or tissues can be derived from optical sections from confocal microscopy.

Analysing (sub)micron dynamics. Rapid image acquisition of fluorescent probes allows imaging (relative) dynamics that can be analysed in kymographs. FRAP studies give insight in fluorescent displacements and allow quantification of protein dynamics.

Polarization microscopy. Biopolymers are made out of molecules arranged in a specific order and as a consequence have a birefringent characteristic. Protein crystals, collagen, cell wall polymers, microtubules or actin filament bundles are examples. Crystallographic and polymer orientation information can be obtained.

Applications

Application areas are in structural analysis, protein interactions, signal transduction, life sciences, single molecule detection, biochip development, food processing, biophysical studies and colloid chemistry.

Ancillary equipment

Ancillary equipment for sample preparation includes (cryo-)microtomes, micro slicer, needle puller, micro manipulation and microinjection.

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In addition to the spinning disk confocal microscope, Wageningen University & Research, Shared Research Facilities offers the use of: