Nikon Eclipse Ti microscope
Skirball 2nd floor

Ideal for:

  • TIRF (Total Internal Reflectance Fluorescence) microscopy with an EMCCD camera or sCMOS camera images a defined volume less than 200 nm deep at the substrate. This allows for exceedingly high contrast, high spatial resolution, and high speed imaging of few molecules. Preparations require a #1.5 glass coverslip and aqueous media.
  • Widefield fluorescence microscopy including long time lapse with multiple fields of view. Also, high speed movies with extra wide field of view.
  • Ratio FRET (Förster Resonance Energy Transfer). Using separate color LEDs for excitation and independently addressable filters for emission, ratio FRET can be done on both live and fixed material.

 

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Instructions:  [Word doc]   [PDF]


This microscope is for widefield fluorescence, TIRF,simple brightfield, and phase contrast. It has automated focus for Z series and precision stage for tiling large areas or repeatably imaging multiple positions. It may be used for live imaging and for fixed samples.

A few of the specifications:

  • Objectives: Plan Fluor 10x/0.3 Ph1, Plan Apo 20x/0.75 DIC, S Plan Fluor ELWD 40x/0.6 Ph2, Plan Fluor 40x/0.75 DIC, Plan Apo λ 60x/1.4 Oil Ph3 and Apo TIRF 100x/1.49 Oil DIC
  • Widefield excitation by LED at 395, 440, 470, 508, 555, and 640 nm. The 555 nm LED emits in a wide spectral range around 555 nm and may be tailored with narrow pass filters.
  • Emission filters in front of Zyla camera: 435/26; 475/20; 515/30; 540/21; 495/40; 632/60; 700/75.
  • Laser excitation (for TIRF) at 405, 488, 561, 640 nm.
  • Emission filters in front of DU897 camera: 450/50; 525/50; 600/50; 700/75; polarizer.
  • Filter blocks (dichroics): "TIRF-quad"; 86006 CFP/YFP/dsRED; 86012v2 Dapi/FITC/TxRed/Cy5; SRIC (reflection); 89021 GFP/mCherry.

The 60X planapochromat lens is the most versatile for most high resolution imaging. With the Zyla camera it oversamples (the equivalent of a 100X lens). Phase contrast makes it ideal for cell culture imaging. It should be used with a #1.5 coverglass

The 100X lens should only be used for TIRF and other specialized imaging where the sample is within a few um of a #1.5 coverglass.

 


Old website here for reference: index_prejuly2018.html