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

Leica Stellaris 5 WLL IR with Resonnant Scanner

Stand	Leica DMi 8 (inverted microscope)
Illumination	–
Lasers	405 nm Diode Pulsed White Laser Line (WLL) – Tunable between 485 nm and 790 nm
Objectives	HC PL APO CS2 10x 0.4 Dry HC PL APO CS2 20x 0.75 Dry HC PL APO CS2 40x 1.30 Oil HC PL APO CS2 63x 1.40 Oil
Stage	Motorized XY Galvo Z-piezo
Contrast	–
CO2 \| T° control	Yes, stage-top incubation (Oko Lab) CO2, Temp. and Humidity control
Detector type	4 HyDs Highly sensitive detectors – Detection from visible to IR wavelengths on the 4 channels Transmitted PMT
Optional modules	FRAP/FRET wizard Lightning (deconvolution) Navigator (Tiling/stitching over huge areas) TauSense (TauContrast, Gating, Separation and Interaction)
Software	LAS X (2022)
Room	CIF 1020.1 GEN

Leica Stellaris 5 with Resonnant Scanner

Stand	Leica DMi 8 (inverted microscope)
Scanner	Galvo scanner (scan speed standard 400 to 2600 Hz) Resonant scanner (28 fps @512x512pixels, 8000 Hz scan speed)
Lasers	405nm, 488nm, 514nm, 561nm, 638nm.
Objectives	HC PL APO CS2 10x 0.4 Dry HC PL APO CS2 20x 0.75 Dry HC PL APO CS2 20x 0.75 IMM HC PL APO CS2 40x 1.30 Oil HC PL APO CS2 63x 1.20 Water
Stage	Motorized XY Galvo Z-piezo
Contrast	NC
CO2 \| T° control	No
Detector type	3 HyD S 1 T-PMT
Optional modules	FRAP/FRET wizard Lightning (deconvolution) Navigator (Tiling/stitching over huge areas)
Software	LAS X (2020) version 4.1.23273
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	LAS X
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]