Edit

Microscopy Facility

Advanced light microscopy support and technology development

X-light V3 Spinning disk

CREST Optics X-light V3 Spinning disk system, Lumencor CELESTA 7-line laser engine, NIKON Ti microscope body, controlled by NIKON NIS

Location: Room 53

System specifications

Spinning disk confocal on an inverted microscope with a 95% quantum efficiency sCMOS camera allowing high-speed confocal imaging that is also well suited for live-cell imaging and high-resolution morphological imaging.

350-900 nm spectral detection range with 5 nm spectral resolution and sub-micron spatial resolution

  • Measuring up to five dimensions, including x, y, z, time and spectrum
  • Capable of multi-dimensional fluorescence qualitative and quantitative measurement, FRET, FRAP, FLIP, FLAP and other photobleaching-related applications
  • Suitable for imaging fixed and live specimen (Enviromental chamber).

Multi-dimension capability: xyz, xzy, xt, xyt, xzt, xyzt, xy, xz, xyt, xyz

Stage:

  • Motorized X, Y stage, resolution 0.1 µm, repeatability +/- 3 µm, max. speed 25 mm/s
  • Motorized z stage (piezo-controlled) with 166.68 µm scanning range

Pinhole size: 50 µm

Software: NIS AR, Version 1.7.0, for image acquisition and processing. Includes NIS denoise.ai and JOBS for building complex acquisition pipelines.

Data structure: 16-bit images (*.tif) as default, also possible 12 bit.

Image size: 2048 x 2048 pixels

Camera

Photometrics Prime BSI, 95% quantum efficiency sCMOS

Lasers

TypeWavelenght (nm)Dyes and use
UV405DAPI, Hoechst
Argon458, 476, 488, 496,  514CyanFP, GFP, Alexa488, CY2
 561RedFPs, Alexa568
 594Alexa 594, mCherry
HeNe633Cy5, Alexa 647
  • An acousto-optical tunable filter (AOTF) controls the intensity of the visible excitation laser light.
  • An acousto-optical beam splitter (AOBS) is used to split excitation light and emission light instead of filter blocks.

Microscope

Leica DMI 6000 CS AFC Bino inverted microscope with motorized XY-stage

Objectives

MagnificationNATypeImmersionCoverslip
20x *0.70PL Apo UVImm/Corr– / 0.17
40x1.25PL Apo UVOil0.17
63x1.20PL Apo UVWater0.14 – 0.18
63x1.4PL Apo UVOil0.17

* The 20x immersion objective for water, glycerine and immersion oil

Fluorescent source

Leica EL6000 External light source for fluorescence excitation (alignment-free, mercury metal halide bulb with an extremely long lifetime of more than 3000 hours)

Fluorescence filter sets

NameExcitation filterEmission filterDichroic
ABP 340 – 380 nmLP 430 nm400 nm
CFPBP 436/20 nmBP 480/40 nm455 nm
GFPBP 470 40BP 525/50 nm580 nm
N2.1BP 515 – 560 nmLP 590 nm580 nm
YFPBP 500/20 nmBP 535/30  nm515 nm

Power up and down procedure

  • Please contact Alvaro Crevenna to schedule a session
  • Report problems on the Log file

Book this resource online (intranet)

Edit