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

Dorigny 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.

Zeiss LSM 880 Airyscan

Stand
  • Axio Observer.Z1

Illumination

  • HXP R 120W

Lasers

  • 405nm

  • 458nm, 488nm, 514nm

  • 561nm

  • 633nm

Objectives

  • Plan Apochromat 10x 0.45 DIC
  • Plan Apochromat 20x 0.8 DIC
  • Plan Apochromat 40x 1.3 Oil DIC
  • Plan Apochromat 63x 1.4 Oil DIC

Stage

  • Motorized XY

  • Piezo Z

Contrast

  • DIC

CO2 | T° control

  • No

Detector type

  • 2 PMT

  • 1 T-PMT
  • 1 Quasar detector (spectral imaging)

  • 1 Airyscan

Optional modules

 

Software

  • Zen black 2.1 SP3

Room

  • GEN-1020.1

Zeiss LSM 710

Stand
  • Axio Observer.Z1

Illumination

  • HXP R 120W

Lasers

  • 405nm

  • 458nm, 488nm, 514nm

  • 561nm

  • 633nm

Objectives

  • Plan Apochromat 10x 0.45 DIC
  • Plan Apochromat 20x 0.8 DIC
  • Plan Neofluar 25x 0.8 Ph2 Multi-immersion
  • Plan Apochromat 40x 1.3 Oil DIC
  • Plan Apochromat 63x 1.4 Oil DIC
  • Plan Apochromat 100x 1.4 Oil DIC

Stage

  • Motorized XY

  • Piezo Z

Contrast

  • DIC

CO2 | T° control

  • No

Detector type

  • 3 PMT

  • 1 T-PMT

Optional modules

 

Software

  • Zen black 2012

Room

  • GEN-1020.1

Zeiss LSM 700

Stand
  • Axio Observer.Z1

Illumination

  • X-Cite 120W PC Q

Lasers

  • 405nm

  • 488nm

  • 555nm

  • 639nm

Objectives

  • Plan Apochromat 10x 0.45 DIC
  • lan Apochromat 25x 0.8 DIC Multi-immersion
  • Plan Neofluar 40x 1.3 Oil DIC
  • C-Apochromat 63x 1.2 Water

Stage

  • Motorized XY

  • Piezo Z

Contrast

  • DIC

CO2 | T° control

  • No

Detector type

  • 2 PMT

  • 1 T-PMT

Optional modules

 

Software

  • Zen 2010

Room

  • BIO-4416

Leica SP8X

Stand
  • Leica DMi 6000

Illumination

  • NC

Lasers

  • 405nm

  • 458nm, 477nm, 488nm, 514nm (Argon)

  • White Laser Line (WLL) – Tunable between 470nm and 670nm

Objectives

  • Plan Fluotar 10x 0.30

  • Plan Apochromat 20x 0.75 CORR

  • Plan Apochromat 40x 1.10 Water CORR

  • Plan Apochromat 63x 1.20 Water CORR

Stage

  • Motorized XY

Contrast

  • NC

CO2 | T° control

  • No

Detector type

  • 3 HyD

  • 1 PMT

  • 1 T-PMT

Optional modules

 

Software

  • Zen 2012

Room

  • BIO-4416

Multi-Photon Confocal

Two-photon excitation microscopy is a fluorescence imaging technique that allows imaging of living tissue up to about one millimeter in depth. It differs from traditional fluorescence microscopy, in which the excitation wavelength is shorter than the emission wavelength, as the wavelengths of the two exciting photons are longer than the wavelength of the resulting emitted light. Two-photon excitation microscopy typically uses near-infrared excitation light which can also excite fluorescent dyes. However, for each excitation, two photons of infrared light are absorbed. Using infrared light minimizes scattering in the tissue. Due to the multiphoton absorption, the background signal is strongly suppressed. Both effects lead to an increased penetration depth for these microscopes. Two-photon excitation can be a superior alternative to confocal microscopy due to its deeper tissue penetration, efficient light detection, and reduced photobleaching.[1][2]

Zeiss LSM 710 NLO

Stand
  • Axio Examiner.Z1

Illumination

  • HXP R 120W

Lasers

1 Photon

  • 405nm

  • 458nm, 488nm, 514nm

  • 561nm

  • 633nm

2 Photon

  • Coherent Chameleon Ultra II IR laser (tunable 690nm to 1040nm)

Objectives

  • Plan Apochromat 20x 1.0 Water
  • Plan Apochromat 40x 1.0 Water DIC 

Stage

  • Motorized XY

Contrast

  • DIC

CO2 | T° control

  • No

Detector type

Internal

  • 2 PMT

  • 1 T-PMT
  • 1 Quasar detector (spectral imaging)

External

  • 2 NDD PMT

Optional modules

 

Software

  • Zen 2010

Room

  • GEN-1020.3

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

  • GEN-1020.2

Laser-capture Microdissection

Laser-capture microdissection (LCM) is a method to procure subpopulations of tissue cells under direct microscopic visualization. LCM technology can harvest the cells of interest directly or can isolate specific cells by cutting away unwanted cells to give histologically pure enriched cell populations. A variety of downstream applications exist: DNA genotyping and loss-of-heterozygosity (LOH) analysis, RNA transcript profiling, cDNA library generation, proteomics discovery and signal-pathway profiling. The total time required to carry out this protocol is typically 1–1.5 h.[3]

MMI CellCut | Olympus IX71 inverted microsocope

Widefield Illumination

  • HBO

Lasers

Laser cutting

  • Solid state UV

Optical tweezer

  • IR laser

Objectives

  • 4x

  • 10x

  • 20x

  • 40x

  • 63x

  • 100x Oil immersion

Stage

  • Motorized XY

Contrast

  • DIC

CO2 | T° control

  • No

Detector type

  •  

Optional modules

  • Wacom Pen Screen

Software

  •  

Room

  • GEN-1020.5

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.

Nikon lab microscope

Stereomicroscopy

The stereo, stereoscopic or dissecting microscope is an optical microscope variant designed for low magnification observation of a sample, typically using light reflected from the surface of an object rather than transmitted through it. The instrument uses two separate optical paths with two objectives and eyepieces to provide slightly different viewing angles to the left and right eyes. This arrangement produces a three-dimensional visualization of the sample being examined.[1] Stereomicroscopy overlaps macrophotography for recording and examining solid samples with complex surface topography, where a three-dimensional view is needed for analyzing the detail.

Nikon SMZ25 Stereomicroscope | Macroscope

Stand
  • Nikon SMZ-25, Upright

Illumination

  • Bright field from the base
  • Fluorescence

Filters

  • DAPI
  • GFP
  • TXR

Objectives

  • Plan Apochromat 0.5x SHR WD 71mm
  • Plan Apochromat 1x SHR WD 60mm
  • Plan Apochromat 2x SHR WD 20mm
  • Motorized 25x zoom

Stage

  • Manual XY

  • Motorized Z

Contrast

  • Brightfield + Fluorescence

CO2 | T° control

  • No

Cameras

  • Nikon DS-Ri2 16 Mpx color camera

Optional modules

  • Extended Depth of Field
  • JOBS
  • Multi-position

Software

  • Nikon NIS-BR 4.20

Room

  • GEN-1020.5

Leica Macroscope

Image Processing

[Under construction]