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CIF Coordinator: jean-yves.chatton@unil.ch

Agora Instruments

Acquisition systems

Confocal

Confocal microscopy, most frequently confocal laser scanning microscopy (CLSM) or laser confocal scanning microscopy (LCSM), is an optical imaging technique for increasing optical resolution and contrast of a micrograph by means of using a spatial pinhole to block out-of-focus light in image formation.[1] Capturing multiple two-dimensional images at different depths in a sample enables the reconstruction of three-dimensional structures (a process known as optical sectioning) within an object.

Olympus FluoView 3000

Illumination

  • LED; pE-300white; CoolLED

Camera

Olympus DP80

Sensor:CCD color
Number of pixels:4080 x 3072
Chip area: 8.8 x 6.6 mm
Pixel chip size: 6.45 x 6.45 µm
Dynamic range: 42 bit (colour)

Lasers

  • 405nm

  • 488nm

  • 561nm

  • 640nm

Objectives

  • Plan Apochromat 1.25x/0.04 WD 5.0mm no contrast dry

  • Plan Apochromat 10x/0.4WD 3.1mm DIC dry

  • Plan Apochromat 20x/0.75 WD 0.6mm DIC dry

  • Plan Fluorite 40x/1.3 WD 0.2mm DIC oil

  • Plan Apochromat 60x/1.42 WD 0.15mm DIC oil

Stage

  • Motorized

Contrast

  • DIC

CO2 | T° control

  • No

Detector type

  • 1x T-PMT

  • 4x GaAsP

Optional modules

 

Software

  • CellSens

  • FluoView 31S-SW

Room

  • B25A.03.201

Spinning Disk | Time-Lapse

Spinning-disk (Nipkow disk) confocal microscopes use 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 phototoxicity and photobleaching of a sample often making it the preferred system for imaging live cells or organisms.

Nikon Ti2 | Yokogawa CSU-W1

Widefield LED Illumination

Lumencore Spectra X LED

  • 395nm

  • 440nm

  • 470nm

  • 508nm

  • 555nm

  • 640nm

Filter Cubes

  • Quintuple multiband filtercube for DAPI/FITC/TRITC/Cy5/Cy7 (32mm)
  • DAPI-5060C Filter Cube (32mm)
  • GFP-4050B Filter Cube (32mm)
  • Cy3-4040C Filter Cube (32mm)
  • Cy5-4040C Filter Cube (32mm)

Spinning Disk laser excitation

Omicron modified LightHUB+

  • 405nm
  • 488nm

  • 561nm
  • 638nm

Objectives

  • CFI Plan Apochromat Lambda 10X, N.A. 0.45, W.D. 4.0mm
  • CFI Plan Apochromat Lambda 20X, N.A. 0.75, W.D. 1.0mm, spring-loaded
  • CFI Plan Apochromat Lambda 40XC N.A. 0.95, W.D. 0.21mm, Spring-loaded, Cover glass correction: 0.11-0.23mm
  • CFI Plan Fluor 40x Oil N.A. 1.30, W.D. 0.2mm, Spring-loaded
  • CFI Plan Apochromat Lambda 60X Oil, N.A1.40, W.D. 0.13mm

Stage

  • Motorized

  • Piezo (Z direction)
  • Nikon PFS (Perfect Focus System)

Contrast

  • DIC

CO2 | T° control

  • Yes [Full enclosure]

Cameras

  • 2x Photometrics Prime 95B

  • 1x Nikon DS-Qi2

Optional modules

  • JOBS
  • General Analysis 3
  • Deconvolution

Software

  • Nikon NIS-Elements AR 5.0

Room

  • B25A.03.201

Widefield | Fluorescence

The majority of fluorescence microscopes, especially those used in the life sciences, are of the epifluorescence design shown in the diagram. Light of the excitation wavelength illuminates the specimen through the objective lens. The fluorescence emitted by the specimen is focused to the detector by the same objective that is used for the excitation which for greater resolution will need objective lens with higher numerical aperture. Since most of the excitation light is transmitted through the specimen, only reflected excitatory light reaches the objective together with the emitted light and the epifluorescence method therefore gives a high signal-to-noise ratio. The dichroic beamsplitter acts as a wavelength specific filter, transmitting fluoresced light through to the eyepiece or detector, but reflecting any remaining excitation light back towards the source.

Zeiss | Widefield

 

Stage
  • Zeiss Axio Observer Z1, Inverted

Illumination

  • X-Cite

Filter Cubes

  • DAPI
  • GFP
  • Rhodamin
  • Cy5

Objectives

  • Plan Apochromat 10X, N.A. 0.45 DIC
  • Plan Apochromat 20X, N.A. 0.8 DIC
  • EC Plan Neofluar 40X, N.A. 1.3 Oil DIC
  • Plan Apochromat 63X, N.A. 1.4 Oil DIC

Stage

  • Motorized

Contrast

  • DIC

CO2 | T° control

  • No | No

Cameras

  • Axiocam 506 color
  • Axiocam 506 mono

Optional modules

  • Tile (Image stitching)

Software

  • Zeiss Zen 2.6 Blue edition

Room

  • B25A.03.201

Image Processing

[Under construction]