Room: S03 V04 F54

Main Applications

• confocal imaging (Spinning Disk), optical sectioning
• fast multi-dimensional live cell + fixed specimen imaging
• multi-position acquisition
• FRAP, Photoactivation / Photoconversion
• TIRF
• epi
• DIC

Funding

Andor/Nikon Spinning Disk confocal and TIRF microscope was funded by the Deutsche Forschungsgemeinschaft (DFG, German Research Foundation) – Project-ID 181168673.

Technical Specifications

Microscope and Accessories

• inverted Nikon Ti-E
• Perfect Focus System PFS
• motorized xy-stage
• Prior Scientific NanoScanZ Piezo stage
• external fluorescence lamp Nikon Intensilight
• Halogen lamp for transmitted light
• EMBLEM microscope incubator box with control device (temperature & CO2 control for live-cell imaging)

Lasers

Andor REVOLUTION 500 series AOTF laser modulator and combiner unit, suitable for launch into CSU/CSUX, TIRF or FRAPPA

• Diode Laser cw 405 nm (100 mW)
• Diode Laser cw 488 nm (50 mW)
• Diode Laser cw 561 nm (50 mW)
• Diode Laser cw 640 nm (100 mW)

Dual Spinning Disk Confocal Scanner

• Yokogawa CSU-X1 Spinning Disk unit, max. 5000 rpm disk spin speed/1000 frames per second
• Andor Revolution FRAP and Photo Activation illumination system (FRAPPA)

Cameras

• Andor Technology iXon DU-897 monochrome backilluminated EMCCD camera, 512 x 512 imaging array, 16 x 16-µm pixels, 14-bit digitization (for Spinning Disk, TIRF)
• Andor Technology Clara camera interline-transfer CCD camera, 1392 x 1040 imaging array, 6.45 x 6.45-µm pixels, 14-bit digitization (for widefield, TIRF)

Objectives

Fluorescence filters & Dichroics

for confocal imaging (emission discrimination and filters):

• CSU Quad Dichroic mirror set for laser wavelength 405 nm, 488 nm, 561 nm, 640 nm
• single band emission filters:
  • DAPI: BP 447/60 nm (Semrock BrightLine)
  • RFP: BP 617/73 nm (Semrock BrightLine)
  • Cy5: BP 685/40 nm (Semrock BrightLine)
• dual band emission filters:
• GFP/RFP: BP 512/23 nm + BP 630/91 nm (Semrock BrightLine)
• GFP/Cy5: BP 538/50 nm + BP 685/45 nm (Semrock BrightLine)

for TIRF imaging (488 nm and 561 nm only):

• Imaging with Andor iXON DU-897
  • TIRF Dual Line Beamsplitter zt 488/561rpc (use together with emission filters of CSU)
• Imaging with Andor Clara
  • GFP: Beamsplitter zt 491 RDCXT, BrightLine HC 525/45
  • RFP: Beamsplitter zt 561 RDCXT, BrightLine HC 509/54

for widefield imaging (filter sets):

• DAPI: EX 340-380, DM 400, BA 435-485
• FITC: EX 465-495, DM 505, BA 515-555
• TRITC: EX 540/25, DM 565, BA 605/55

Software

Andor IQ3