Widefield Microscopy

The General Instrumentation Department hosts the following widefield microscopes.

Leica Thunder (room HG01.228)

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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)

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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)

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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

Leica DMIRE2 (room HG01.228)

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Microscope:

  • Inverted DMIRE2 microscope for bright-field, and epi-fluorescence

Filters:

  • A for UV excitation BP340-380, DM 400, LP 425
  • N2.1 for green excitation BP 515-560, DM 580, LP 590
  • I3 for blue excitation BP 450-490, DM 510, LP 515

Objectives:

  • 10x/0.3 Air
  • 20x/0.7 Air
  • 63x/1.3 Glycerol
  • 63x/1.4 Oil

Detection: Photometrics CoolSnap K4 monochrome CCD camera 2048 x 2048 imaging array, 7.4 x 7.4-um pixels, 15.16 x 15.16-mm imaging area (optically centered)

Software: uManager 1.4 controlling the camera, the stage the filters wheel and transmitted light.

Leitz HM-Pol (room HG01.235)

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Microscope:

  • Upright polarization microscope with a rotating stage and an optional retardation plate.

Objectives:

  • 4x/0.12 Air
  • 25x/0.35 Air
  • 40x/0.65 Air

Leica MZFLIII Stereoscope (room HG01.228)

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Microscope: Stereoscope with a continual zoom magnification (from 0.8x to 10x) is appropriate for object of about 100 µm to a few mm. It can be used in transmission, epi-illumination and dark field mode. For the epi-illumination one can choose between two light spots or an annular light source (less shadow).

Detection:  Coolsnap 5M (Photometrics) color camera operated with the Micro-Manager Software. (Resolution: 2580 x 1944 active pixels; Pixel Size : 3.4 µm x 3.4 µm)