HomeAboutNewsCombined epifluorescence and atomic force microscopy now available (Sept 2016)

Combined epifluorescence and atomic force microscopy now available (Sept 2016)

CCMR’s popular Asylum MFP-3D-BIO atomic force scanning probe microscope is built atop an Olympus IX71 inverted optical microscope. Epifluorescence microscopy is now possible with the addition of a SOLA excitation light source, a collection of epifluorescence filter cubes, a variety of objective lenses, and a low-noise Andor Zyla 4.2 Scientific CMOS imaging detector. The epifluorescence imaging capability is now available for general use.

CCMR currently provides three filter cubes, in addition to the original set supporting standard bright-field imaging:

Filter cube Excitation range Dichroic cut-on wavelength Emission range
Olympus U-MBF3 Visible N/A Visible
Chroma 39007 590-640

(orange-red)

654

(red)

670-720

(red-infrared)

Olympus U-MNIBA 470-495

(blue)

505

(green)

510-550

(green)

Olympus U-MNG 530-550

(green)

570

(yellow)

590+

(yellow-orange-red)

These three filters were used to generate a composite red/green/blue image of an array of 10-15 um spheres tagged with three different fluorophores. The detector can operate in 12- or 16-bit mode, with low read noise and dark current, allowing detection of even weakly fluorescing samples.

Sphero Combined 10x

We currently have room to accommodate two additional filter cubes, and welcome suggestions for additional filter cubes that are necessary for your research. The cubes we already have will support the following:

  • Carbohydrate/protein modification: Alexa Fluor 488/532/568/647, FITC, Oregon Green 488, PE-B, PE-R, RITC
  • Protein modification: Cy5, Eosin-isothiocyanate, FluorX, Resorufin
  • Nuclei, nucleic acid: Draq5, Ethidium Bromide, Pararosaniline-Feulgen, PI, SYTOX Green, YO PRO 1, YOYO 1
  • Live/dead cell staining: BCECF/AM, Calcein/AM, CFDA, FDA, Ethidium Bromide, Ethidium homodimer-1, PI
  • Membranes: DiL, Merocyanine 540
  • Organelle: BODIPY FL Ceramide, DiO C6(3), JC-1, MitoFluor Green, Rodamine 123
  • Bone: Acidic Fuchsin, Xylenol Orange
  • Nerve cell tracer: Evans Blue
  • Actin fibers: Rhodamine-Phalloidin
  • FISH probe labeling: Spectrum Green, Spectrum Orange

If you have questions or are interested in using the microscope, please contact Darren Dale (darren.dale@cornell.edu) or Steve Kriske (kriske@cornell.edu).

 

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