Widefield Microscopy

The General Instrumentation Department hosts the following widefield microscopes.

Leica Thunder (room HG01.228)

Afbeelding2

Microscope:

  • Automated DMi8 (inverted) with hardware autofocus (AFC) and motorized polarizer.

Excitation Lines: Lumencor 4-line LED with the following wavelengths

  • 395nm
  • 488nm
  • 561nm
  • 638nm

Emission Selection: EFW - external filter wheel slider for high speed acquisition. Filterset– quad cube for imaging (DAPI, FITC, TRITC, Cy5) including filter wheel for EFW

Detection: Leica DFC9000 GTC

  • 2 MP sCMOS camera, 6.5 μm pixel size (19 mm sensor diagonal)
  • 82% quantum efficiency, dark current of 0.14e-/p/sec
  • ~90 frames per second acquisition speed
  • extended dynamic range of 33000:1

Objectives:

  • 20x/0.8 Air
  • 63x/1.2 Water

Stage: High speed/precision quantum stage

Environmental Control: Incubator with temperature controller

Software Modules:

  • LAS X 3D for 3D rendering of acquired volumes
  • “Navigator” for large tile scans
  • “Thunder” for instant or volume computational clearing and improvement of resolution

Leica DM2500 (room HG01.235)

Afbeelding3

Microscope:

  • Upright manual microscope for bright-field, phase-contrast, dark-field and epi-fluorescence

Filters:

  • Transmission (BF, Ph, DF), no filters
  • Ultraviolet (DAPI, aniline blue), Filter A, Exc. BP 320-380, DM 400, Em.LP 425
  • Blue (Fluorescein, Alexa 488), Filter I3, Exc. BP 450-490, DM 510, Em. LP 515
  • Green (Rhodamine, Alexa 561, Cy3), Filter N2.1, Exc. BP 515-560, DM 580, Em. LP 590
  • Red (Cy5), Filter Y5 ET, Exc. BP 590-650, DM 660, Em.BP700/75

Objectives:

  • 5x/0.07 Air
  • 10x/0.3 Air
  • 20x/0.5 Air
  • 40x/0.75 Air
  • 100x/1.3 Oil

Detection: Leica DFC7000 T controlled by the LAS X software

  • 2.8 MP CCD
  • Pixel size 4.54 x 4.54 μm
  • 1920 x 1440 pixels
  • 8/12 bit with 16-bit A/D converter
  • 40 fps (full frame) / 123 fps (5 x 5 binning)

Leica DMRA (room HG01.235)

Afbeelding4

Microscope:

  • Upright manual microscope for bright-field, phase-contrast and epi-fluorescence

Filters:

  • A: BP340-380 - 400 - LP425
  • S-Aqua: D436 / 20x - 455 - D480 / 30
  • GFP ET: 47- / 40 - 495 - 525/50
  • N2.1: BP515-560 - 580 - LP590
  • Y5: BP620 / 60 - 660 - BP700 / 75
  • Cyan GFP V2
  • GFP BP Yellow

Objectives:

  • 6x/0.05 Air
  • 5x/0.07 Air
  • 5x/0.15 Air
  • 10x/0.3 Air
  • 20x/0.7 Air
  • 40x/0.5 Air

Detection: Leica DFC340FX camera operated with the LAS AF software