Microscopy Assets
Microscopes and related equipment:
- Two photon/confocal microscope – custom built
- Nikon TE2000-U stand.
- Halogen condenser illumination.
- Mercury arc epifluorescence.
- Variety of fluorescence cubes.
- Objectives:
- Nikon 60x/1.27NA Plan Apo IR water immersion with DIC.
- 40x/0.6NA Plan Fluor with phase contrast.
- 20x/0.75NA Plan Fluor ELWD with DIC.
- Physik Instrumente fast piezo focuser on 60x objective.
- Argon ion laser (Spectra-Physics).
- Titanium:Sapphire laser: Mai Tai HP (Spectra-Physics).
- Fast Pockels cell (Conoptics) on Mai Tai beam path.
- Two Avalanche photodiode detectors (Perkin Elmer) – single photon counting modules.
- Two Hybrid detectors (Hamamatsu/Becker and Hickl).
- Becker and Hickl fluorescence lifetime module.
- x-y galvanometer mirrors (Cambridge Technology).
- Custom LabVIEW-based instrument control and acquisition software and hardware (National Instruments).
- Vibration dampened optical bench with optics and optomechanical components (various vendors).
- Custom applications:
- Quantitative subcellular protein distribution analysis
- 2 photon FRAPa and FRAPb
- 2 photon FLIM-based FRET
- 2 photon FLIM-based polarization anisotropy microscopy
- Fluorescence correlation spectroscopy
- sptPALM and STORM microscope (super resolution).
- Nikon Ti2 stand.
- Objectives:
- SR HP Apo TIRF 100x/1.49 Oil.
- 20x/0.75NA Plan Apo.
- LED condenser illumination.
- LED EXFO epifluorescence.
- 3D N-STORM module with high power 4 laser system.
- ILAS2 ring TIRF illumination.
- Ixon Ultra 892 EMCCD camera.
- Prime BSI sCMOS camera.
- Live cell incubation chamber – okolab.
- NIS-Elements Advanced Research
- AI module.
- 3D super resolution analysis.
- AI-mediated automated acquisition.
- Custom applications:
- sptPALM for high density mapping of single molecule dynamics in live cells.
- 3D sptPALM under development.
- Qdot-based single molecule tracking in live cells.
Microscopes and related equipment:
- Leica TIRF/dSTORM microscope (super resolution)
- LED widefield illumination
- Solid state 120-150 mW 405, 488, 561, and 651 lasers
- Standard fluorescent cubes and filters (for wavelengths above)
- Objectives:
- 20x air
- 40x oil pl apo 1.25 NA DIC
- 63x pl apo 1.47 NA TIRF/DIC, oil (temperature correction)
- 100x pl apo 1.47 NA TIRF/DIC, oil (temperature correction)
- 163x pl apo 1.45 NA TIRF/DIC, oil (temperature correction)
- DIC and polarization setup
- Motorized stage
- LASX software with STORM module
- Andor iXon EMCCD camera
- Antivibration table
- Autofocus
- Advanced peristaltic pump
- Custom microfluidics devices
- OKO labs heater (stage insert and objective heater) – no gas
Applications:
- Standard epifluorescence
- TIRF/HiLo (VAEM) for live or fixed cells
- TIRF “biochemistry on a coverglass” reconstitution experiments
- SNAP- Halo-tag based single molecule tracking in live cells and in vitro reconstitution assays
- Nikon SoRa spinning disk confocal and deconvolution system (super resolution)
- SOLA LED widefield illumination
- Solid state Solid state 100 mW 405, 488, 561, and 651 lasers
- Photometrics Prime sCMOS camera
- Objectives
- 20x pl apo 0.75 NA
- 60x pl apo 1.4 NA
- Piezo stage
- Motorized turret and stage
- NIS-elements software
- SoRa deconvolution software upgrade
- Antivibration table
- Yokogawa CSU-W1
Applications:
- Super resolution spinning disk confocal (40-50 nM resolution)
- Single molecule tracking in cells
Extremely fast 4D (Z stack + time) movies
The lab houses a TIRF microscope that belongs to the CDB department.
This is a Nikon TE2000 microscope equipped for live cell imaging (with a temperature-controlled environmental chamber).
The following imaging modalities and objectives are available: IRM, DIC (60X and 20X), phase contrast (10X), epifluorescence (20X, 60X, 100X), and 4-color TIRF (60X or 100X). The following TIRF lasers are available: 405 nm, 488 nm, 561 nm, and 640 nm.
The microscope is equipped with the motorized Z-position control, Perfect Focus, and with a Prime 95B cameras and is controlled by a dual monitor computer using Nikon Elements software. The microscope is equipped with filter sets for multicolor TIRF, DAPI, TRITC, GFP, and CY5.
Zeiss LSM 780 confocal
- The LSM780 is an inverted microscope with 4 lasers, 3 detectors, an environmental chamber, and mercury and halogen light sources. The microscope has 6 excitation color capabilities, and 3 independent fluorescence imaging channels including one spectral, and one transmission PMT. The system is configurable for simultaneous and/or sequential multicolor acquisition.
Leica SP8 with STED
1.Confocal Microscopy
- Key Features:
- UV Laser 405 nm, White light laser WLL2 470-670 nm.
- Detection objectives: 10x dry, 20x dry, 40x oil, 63X oil.
- Five spectral detector channels: two PMTs and three HyDs.
- Tandem scanner (8kHz).
- XY Scanning Stage.
- HyVolution2 SuperResolution package
2. STED (stimulated emission depletion) 3X nanoscopy
This is a super-resolution technique that overcomes diffraction limited resolution. With multiple STED laser lines of at 592 nm, 660 nm and the pulsed laser at 775 nm, STED can reach a resolution below 30 nm.
- Key Features:
- UV Laser 405 nm, White light laser WLL2 470-670 nm.
- STED Laser: 592 nm, 660 nm and 775 nm.
- STED objectives: 93x glycerol and 100x oil.
- Five spectral detector channels: two PMTs and three HyDs.
- Tandem scanner (8kHz).
- XY Scanning Stage.
- HyVolution2 for STED.
- Select a confocal geometry optimal for your assay and throughput needs
- Achieve excellent image quality without sacrificing throughput via our unique optical path technology
- Acquire statistically relevant data quickly with an advanced scientific CMOS detector, enabling >3 log dynamic range
- Large field of view enables whole-well imaging
- Expand your research capabilities with transmitted light, liquid handling, and environmental options
Confocal technology at the speed of widefield imaging:
- Capture an entire well of a 384‑well plate with a single image at 4X magnification
- Capture four wells of a 1536- well plate in a single image at 4X magnification
- Throughput of >160K wells/day confocal, >200K wells/day widefield 2 ImageXpress
- JEM-2100 (JEOL) 80-200 kV cryo configuration
- LaB6 electron source
- Gatan 626 cryo stage
- TVIPS F415-MP 4096x4096 pixel slow scan CCD camera, 15 µm pixel size, 6.1 x 6.1 cm imaging area
- TVIPS F114-NX 1024x1024 pixel fast scan CCD camera, 14 µm pixel size, up to 26 frames/sec for life imaging
- EMMENU software for manual and automatic image collection
Leica SP8 confocal microscopy: (WH-108):
Leica's latest confocal system with three channel spectral detectors and HyVolution software that combines super-sensitive HyD detection with SVI Huygens deconvolution, allowing lateral resolution down to 140 nm compared with 200-250 nm for standard confocal microscopy.
Key Features:
- UV Laser 405 nm, VIS Laser 488 nm, 552 nm and 638 nm.
- Detection objectives: 10x dry, 20x dry and 63x Oil.
- Three spectral detector channels: two PMTs and one HyD.
- XY Scanning Stage.
- HyVolution2 SuperResolution package.
- Select a confocal geometry optimal for your assay and throughput needs
- Achieve excellent image quality without sacrificing throughput via our unique optical path technology
- Acquire statistically relevant data quickly with an advanced scientific CMOS detector, enabling >3 log dynamic range
- Large field of view enables whole-well imaging
- Expand your research capabilities with transmitted light, liquid handling, and environmental options
Confocal technology at the speed of widefield imaging:
- Capture an entire well of a 384‑well plate with a single image at 4X magnification
- Capture four wells of a 1536- well plate in a single image at 4X magnification
- Throughput of >160K wells/day confocal, >200K wells/day widefield 2 ImageXpress