Universitat Pompeu Fabra
Core Facilities

Confocal Microscopy

Leica DM IRBE inverted research microscope and optical outfit
Inverted microscope for transmitted light brightfield and fluorescence of incident light.
- Tube optics HC with 1x lens and Bertrand lens for phase contrast and Nomarski (or differential interference contrast (DIC)).
- Motorized macro/micro focusing.
- Eyepieces HC plan 10x22 BR.M.
- Condenser S23 (0.53 NA).
- Light source: mercury lamp with 50 W bulb.
- Fluorescence filter blocks: I3 (blue excitation BP 450-490 nm, dichromatic 510, emission LP 515 nm; for FITC, Cy2); N2.1 (green excitation BP 515-560 nm, dichromatic 580, LP emission 590 nm; for TRITC, Cy3) and a triple filter B/G/R (BP420/30nm, dichromatic 415, BP emission 465/20).
- Objectives: HC PL Fluotar 10x /0.30 Ph1; HC PL Fluotar 20x /0.50 Ph2; HCX PL APO 40x /1.25 Oil Ph3 CS; HCX PL APO 63x /1.32 Oil Ph3 CS; HCX PL APO 100x /1,40-0.70 Oil CS.
- Contrast techniques: interference contrast for 40x and 63x. Phase contrast for 10x, 20x, 40x and 63x.

Leica TCS SP2 spectral confocal module
- Lasers: Argon (458 nm 5 mW, 476 nm/ 5 mW, 488 nm 20 mW, 514 nm/ 20 mW), HeNe (543 nm/ 1.2 mW) and HeNe (633 nm/ 10 mW). Optical fibre connection.
- Range of detection spectrum 400-850 nm.
- Line frequency up to 2,000 lines/sec, 3 fps (frames per second) at 512x 512; 25 fps at 512x 31 pixels.
- Zoom 1....32 x
- Diagonal of the scanned area 22 mm.
- Scan rotation: -5...+95 degrees.
- Detection pinhole diameter, adjustable and computer-controlled.
- AOTF (Acousto-optical tuneable filters) 6 channels. Allows simultaneous control of 6 laser lines. Users can change laser lines quickly and control their intensity individually. - 3 simultaneous detection channels for confocal reflected light and/or fluorescence. Continually adjustable spectrum centre and width, independent for each channel.
- Anti-vibration table for inverted microscope, with steel base and granite platform.
- Resolution: 4,096 x 4,096 pixels x 12 bits.

High-precision focus control system.
Galvanometer driven stage with z axis control and a 40 nm resolution.

Transmitted light detection channel. Non-confocal.
Transmitted light detector to record brightfield images. Non-confocal images.

Complete incubation system
Incubation chamber with gas and temperature control, thermal adapter for petri dishes (35 or 60 mm Ø), Bachofer perfusion chambers, and an adapter (Molecular Probes atto-fluor) that can be fitted to this system.

System software
TCS SP2 on-line and electronic work station
The system is controlled by an Intel PIII processor (600 MHz and 256 MB RAM) that works with Windows NT4.0 operating system. It has two 21" real-colour monitors with a total resolution of 2,560 x 1,024 pixels. CD reader/writer.
The following software is installed on this processor:
- LCS software to control browsing, acquisition of multidimensional series, display, two-dimensional processing, quantification and documentation of images.
- LCS 3D software for reconstructing images in three dimensions.
- LCS Multicolor software analysis of two- and three-channel colocalization, 2D and 3D cytofluorograms, histograms...
- LCS Physiology software, enabling the quantification and measurement of ions in living cells...
Off-line work station
Includes the following applications:
- LCS software, enabling display, two-dimensional processing, quantification and documentation of images.
- LCS 3D software for three-dimensional reconstruction of images.
- LCS Multicolour software for colocalization analysis.

DM RB vertical research microscope fitted to Leica DC300 F digital camera
Vertical research microscope with transmitted light brightfield and incident light fluorescence.
- Optical: Objectives: 10x 0.30 HC PL Fluorar, 20x 0.70 HC PLAN APO, 40x 1.25 Oil PLAN APO, 100x 1.30 HCX PL Fluotar.
-Contrast techniques: interference contrast for 20x and 40x.
- Leica DC300F digital and electronic system camera
RGB camera with colour filter, 1,300 x 1,030 pixels and 6.7x 6.7 mm pixel measurement. Progressive scanning, exposure time: 0.3 ms- 60s.
The system is controlled by an Intel Pentium 4 processor and Windows 2000 operating system. They also have a CD reader/writer.

Cell manipulation and microinjection system.
Eppendorf InjectMan (adherent cell microinjection).
Eppendorf TransferMan NK (suspension cell micromanipulation).
Eppendorf FemtoJet (enables reproducible microinjections).
Eppendorf CellTram Air (enables suspension cell manipulation).

Vertical pipette puller
Narishige vertical pipette puller, PD-10 model.

Last updated 24-02-2009
© Universitat Pompeu Fabra, Barcelona