Confocal & Specialized Microscopy Microscopes and Supporting Equipment

Choose by imaging method
2-photon excitation
Histology (color brightfield imaging)
Laser capture microdissection, photoablation, targeted DNA damage
LED fluorescence excitation
Resonant scanning confocal imaging
Scanning confocal imaging
Spectral confocal imaging
Spinning-disk confocal imaging
Super-resolution localization microscopy (STORM/fPALM)
Super-resolution structured illumination microscopy (SIM)
Total internal reflection fluorescence (TIRF)
Wide-field fluorescence
or choose an instrument
Red: Nikon A1 Confocal
Orange: Nikon A1RMP
Yellow: Nikon N-SIM
Green: Nikon N-STORM/TIRF/SD
Blue: Zeiss MicroBeam LCM/Histology

Accessory equipment and live-cell facilities

Red: A1 Confocal Microscope

 

Hepatic fibrosis. R. Schwabe. (Troeger et al., Gastroenterology, 2012)

 
Microscope stand Nikon Ti Eclipse inverted
Features . Motorized XY Stage, allowing mosaic images and multi-point time-lapse imaging
. Autofocus using the Perfect Focus System
. Targeted photobleaching, FRAP, FRET and FLIP capability
Samples Slides, 35 mm round culture dishes, chamber slides, chamber coverslips, multiwell plates.
Fluorescence excitation 405, 488, 561, 638 nm
Compatible fluorophores Similar to DAPI, DyLight 405, FITC, TRITC, Cy5
Objectives Nikon Objectives
Software NIS-Elements

Orange: A1RMP Multiphoton Confocal Microscope

 

Embryonic mouse kidney (2-photon excitation). F. Costantini.

 
Microscope stand Nikon Ti Eclipse inverted
Features . Motorized XY Stage, allowing mosaic images and multi-point time-lapse imaging
. Fast live imaging with resonant scanner and piezoelectric focus
. Spectral unmixing of overlapping fluorophores or autofluorescence
. Autofocus using the Perfect Focus System
. Tokai Hit stage-top incubator and objective heater
. Targeted photobleaching, FRAP, FRET and FLIP capability
. 2-photon microscopy with non-descanned detector: Best for deep (up to 1mm) or long-term imaging
. Spectral fingerprinting, multiphoton excitation fingerprinting, unmixing of multiple fluorophores/autofluorescence sources
Samples Slides, 35 mm round culture dishes, chamber slides, chamber coverslips, multiwell plates
Fluorescence excitation . 1-photon: 458, 488, 514, 561, 639 nm
. 2-photon: Software-tunable Chameleon II laser (680-1040 nm)
Compatible fluorophores . 1-photon: Similar to DAPI, DyLight 405, CFP, FITC, YFP, mOrange, TRITC, Cy5
. 2-photon: Compatible with most red, green and blue fluorophores
Objectives . 1-photon: Nikon Objectives
. 2-photon: 25x/1.10 coverslip-corrected water-immersion IR lens
Software NIS-Elements

Yellow: Structured Illumination Microscope (SIM)

 

Wide-field deconvolution (top) and SIM (bottom) images of meiotic chromosomes in spermatocyte spreads. (Horn et al., doi: 10.1083/jcb.201304004, 2013.)

 
Microscope stand Nikon Ti Eclipse inverted
Features . SIM provides double the resolution of the conventional confocal microscope (lateral ~58-110nm; axial ~200-250nm)
. Autofocus using the Perfect Focus System
. 3D Axial Range: up to 20μm
. Imaging speed: 0.6 sec/frame (2D SIM); 1.0 sec/frame (3D SIM)
. Motorized Stage and Piezo Z
Samples Slides, 35 mm round coverslip-bottom culture dishes, chamber slides, chamber coverslips
Fluorescence excitation 405, 488, 561, 647 nm
Compatible fluorophores DAPI, FITC, TRITC, Cy5
Objectives Nikon Objectives
Software NIS-Elements

Green: N-STORM Stochastic Optical Reconstruction Microscope

 

Comparison of conventional and STORM images of microtubules

Comparison of a cell viewed with epi-fluorescence (left) and TIRFM (right)

Microscope stand Nikon Ti Eclipse inverted
Features . Tokai Hit stage-top incubator and objective heater
. Motorized Stage and Piezo Z
. EMCCD Camera (512x512) (Andor DU-897 and Evolve)
Features:STORM . Allows imaging fixed samples with a lateral resolution of ~20-30nm and an Axial resolution of ~50-60nm
. 2D and 3D STORM capabilities
Features:Spinning-disk confocal imaging . Yokogawa CSU-X1
. Allows very fast imaging of bright samples (>50 frames per second)
. Lower photodamage than scanning confocal imaging
Features:Total Internal Reflection Fluorescence Helpful for imaging events at the cell surface (~100nm) with very good signal/noise.
Samples Slides, 35 mm round culture dishes, chamber slides, chamber coverslips, multiwell plates.
Fluorescence excitation 405, 488, 561, 647 nm
Compatible fluorophores DAPI, FITC, TRITC, Cy5
Objectives For TIRF and STORM, 60x/1.49 and 100x/1.49; for spinning-disk, see Nikon Objectives
Software NIS-Elements

Blue: Wide-field fluorescence microscope with PALM Microbeam IV for laser capture microdissection and microirradiation and color histology imaging

 

Noncontact laser capture microdissection. From zeiss.com.

Recruitment of NKX3.1 to the site of induced DNA damage. E. Gelmann. (Cai et al., Cancer Res., 2010.)

 
Microscope stand Inverted Zeiss AxioObserver.Z1
Features . Automated stage for multi-location time-lapse or mosaic imaging
. Autofocus using hardware (Definite Focus) or software
. Pecon stage-top incubation
Features: Wide-field fluorescence . Bioptechs objective heater
. Fast switching LEDs for multicolor live imaging
. Orca ER cooled CCD camera
Features: Microirradiation and Microdissection • Non-contact laser catapulting for capturing live cells or material from frozen or fixed tissue
• Captured material can be used for isolation of DNA, RNA, or protein, or for cell/tissue culture applications
• 355-nm solid-state laser can also be used for
   o creating localized double-strand breaks in DNA
   o ablating cells or subcellular structures
   o photobleaching or photoactivation
Features: Color histology imaging Axiocam ICc for color imaging and histological staining
Samples Slides, 35 mm round culture dishes, chamber slides, chamber coverslips, multiswell plates
Fluorescence excitation Mercury halide lamp with fluorescence filters; LEDs at 390, 455, 470, 505 and 590 nm
Compatible fluorophores DAPI, FITC, TRITC, CFP, GFP, YFP, RFP (DsRed)
Objectives Zeiss Objectives
Software . Zen Blue
. PALM Robo

Accessories

A lab bench with dissecting microscope, minifuge, vortexer, water bath, and hotplate are available for basic sample preparation. For live-cell imaging experiments, a cell culture incubator with CO2 is available in the main microscope room. We also have a laminar flow hood for sterile tissue culture procedures. Stagetop incubators provide temperature, humidity, and CO2 control during experiments. A syringe pump and peristaltic pump are available for microfluidics and perfusion experiments. The CellAsic ONIX microfluidic system offers the opportunity for stable cell culture with simultaneous high-quality, long-term imaging of mammalian cells, yeast, or bacteria.

Nikon Objectives

Magnification Numerical Aperture Immersion Notes
10x 0.4 air OK for plates
20x 0.75 air high-res, low-mag
25x 1.1 water 2-photon
40x 0.6 air OK for plates
40x 0.75 air highest resolution without oil
40x 1.3 oil  
60x 1.49 oil TIRF, super-resolution
100x 1.45 oil  
100x 1.49 oil TIRF
100x 1.45 oil TIRF, super-resolution

Zeiss Objectives

Magnification Numerical Aperture Immersion Notes
2.5x 0.075 air  
5x 0.16 air  
10x 0.3 air OK for plates
10x 0.5 air high-res, low-mag
10x 0.4 air OK for plates
20x 0.4 air laser capture, OK for plates
20x 0.6 air laser capture, OK for plates
40x 0.75 air highest resolution without oil
40x 1.3 oil  
100x 1.3 oil