03. Leica "Thunder"

Microscope:

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

Excitation Lines: Lumencor SPECTRA-X 7-line LED with the following available wavelengths

• 395 nm
• 440 nm
• 470 nm
• 510 nm
• 550 nm
• 640 nm

Emission Selection:

DFT5 quad-cube for high speed fluorescence imaging (Blue, Green, Red, Far-red, or 100%) enabling specific capture of:

• 420-450 nm (eg. DAPI, Hoechst)
• 506-532 nm (eg. GFP, FITC)
• 581-607 nm (eg. RFP, TRITC, Cy3)
• 666-724 nm (eg. AlexaFluor 633, DRAQ5, Cy5)

CFP filter cube to capture:

• 460-500 nm (e.g. CFP, TFP, Cerulean)

YFP filter cube to capture:

• 520-550 nm (e.g. YFP, mVenus)

Detection: Leica DFC9000 GTC

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

Objectives:

• 5x/0.12 Dry (working distance 14 mm)
• 10x/0.40 Dry (2.74 mm)
• 20x/0.80 Air (0.40 mm)
• 63x/1.20 Water (0.30 mm)
• 63x/1.30 Glycerol (0.28 mm)

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 scan
• “Thunder” for instant or volume computational clearing and improvement of resolution