CTI Technologies

Electron Microscopy

JEOL 1200 EXII TEMSCAN analytical electron microscope. With a maximum kV of 125, the JEOL has the capabilities to do minimum electron dose imaging, and it has a side entry goniometer stage for stereo viewing. The JEOL has recently been enhanced with a new 11 megapixel AMT camera.

FEI Tecnai G² F20 TWIN Field Emission Gun Electron Microscope. The column of this 200 kV scope contains a factory aligned Schottky field emitter and isolated vacuum system for the field emission gun (FEG), diffraction lens, two condenser lenses, the TWIN objective lens with a computerized 5-axes eucentric goniometer, two projector lenses, an intermediate lens and a housing for the camera. The Tecnai G² has the capability to do automated tomography, low dose exposure, fast spot scan, and photomontage. This scope is equipped with an Eagle camera that is sensitive enough for cryo TEM.

Other technologies include a Leica AFS2 Freeze Substitution Unit, 4 Leica Ultramicrotomes (one with Cryo capabilities), a Leica DM 4000 with camera, a Leica IGL Immunostainer, 2 Lynx II automated Tissue Processors, a Leica TP automated Tissue Processor, and several image analysis computers.

Light Microscopy

Zeiss LSM 780 NLO

The LSM 780 NLO is configured for both confocal and multiphoton excitation (MPE) imaging and is best suited for multichannel fluorescence detection in fixed or living specimens. The instrument is mounted on an inverted Axio Observer microscope and fitted with an environmental enclosure for control of temperature and CO2. A variety lenses are available for fluorescence and Differential Interference Contrast (DIC) imaging. The complement of available lasers allows for excitation of fluorophores from the violet through the near IR. Single photon excitation lasers lines are 405nm 458nm, 488nm, 514nm, 561nm and 633nm. The wide range of available excitation wavelength along with the 32-channel GaASP detectors makes this instrument extremely well suited to record fluorescence emission spectra and linearly unmix up to 10 individual fluorophores in a single sample. In addition, a tunable femtosecond pulsed laser is available for MPE. Multiphoton capabilities are best applied to improve depth of imaging penetration into thick specimens and to minimize phototoxic effects of imaging in living cells and tissues. Descanned confocal or multiphoton signal may be collected through internal detectors, however, improved MPE signal collection will be achieved by using the non-descanned detectors (NDDs). The microscope is also equipped with a precision motorized stage, enabling multi-point time-lapse experiments as well as tiling over a large area of the specimen.

Nikon C-series Confocals

The Nikon C series laser scanning confocal microscopes are well suited for multichannel fluorescence detection in living or fixed specimens. These confocals are mounted to Nikon TE2000 inverted microscopes and fitted with incubator enclosures for control of temperature and CO2. A variety lenses are available for fluorescence, Phase Contrast, and Differential Interference Contrast (DIC) imaging. In addition to confocal imaging, both systems may be configured for camera based collection of fluorescence or brightfield images.

The C2 is configured with four laser lines (404nm, 488nm, 561nm, and 638nm) optimized for excitation of the most commonly used fluorophores. The integrated Perfect Focus System (PFS), a dynamic focus correction system, tracks the position of the coverslip with infrared light and corrects for any drift in focus with millisecond temporal resolution, eliminating focus drift during long term timelapse imaging experiments. An integrated, motorized stage allows for collection of multipoint z-stacks and for tiling of adjacent fields.

The C1Si is configured with a 3-line Argon ion laser (458nm, 488nm, 514nm) in addition to the 405nm, 561nm, and 638nm lasers. The full complement of laser lines provides the best results from the spectral detection feature. Fluorescence emission spectra can be obtained for any signal on a pixel-by-pixel basis, which is useful for separating closely overlapping fluorophores, distinguishing specific signal from background autofluorescence, and performing spectral FRET.

Spinning Disk Confocal Imaging Systems

The Marianas systems, Marianas 1 and Marianas 2, are multimodal imaging workstations featuring CSU-X spinning disk confocal heads. Each is configured on a Zeiss Axio Observer microscope with 6 diode lasers providing the following wavelengths: 405nm, 442nm, 488nm, 515nm, 561nm, and 642nm. Designed primarily for live cell imaging experiments, these systems are equipped with full incubator enclosures, affording excellent cell viability during long term time-lapse experiments. The spinning disk confocal approach is advantageous in that it offers decreased photobleaching and phototoxicity as well as increased sensitivity and speed over standard point scanning confocal methods.

Both Marianas 1 and Marianas 2 include Vector scanning accessories, which permit exact control of laser positioning to produce areas of photobleaching or photoactivation. Coupled with spinning disk confocal imaging, highly dynamic processes may be recorded. Additionally, both systems hare equipped with Definite Focus Systems, which actively maintain focus during time-lapse imaging experiments. Marianas 1 has been configured for Total Internal Reflections Fluorescence (TIRF) imaging. This imaging modality produces a thin optical section at the specimen-coverslip interface, resulting in significantly improved signal-to-noise (S/N) and z-resolution for events occurring at membrane surfaces such as vesicle trafficking. TIRF is also the basis for STORM and PALM super resolution microscopy. This instrument is also configured for single photon excitation Fluorescence Correlation Microscopy (FCS) and Fluorescence Lifetime Imaging.

Zeiss Elyra PS.1

For projects requiring enhanced resolution, the Elyra PS.1 platform integrates superresolution modalities of PALM (Photoactivated Light Microscopy), STORM (Optical Reconstruction Microscopy), and SIM (Structured Illumination Microscopy). All modalities share the same laser launch delivering 405nm, 488nm, 561nm and 640nm light for excitation of fluorophores. SIM images are a product of algorithm calculations on multiple images acquired through a series of grid patterns. A two-fold improvement in resolution can be achieved in all three dimensions. SIM imaging can be performed with standard fluorophores. Alternatively, the high-precision localization microscopy methods of PALM and STORM can be employed to achieve ten times improvement in resolution over standard fluorescence imaging methods. PALM and STORM require the use of special fluorophores; protocols for preparation of samples are available through the CTIC-LM. A Tokai Hit stage-top incubator is available for control of specimen temperature and CO2.

Zeiss Lightsheet Z.1

Sheet plane illumination imaging is designed for multiview imaging of larger specimens unconstrained by coverslips and slides. A highly 3-dimensional specimen is suspended in a sheet of laser illumination for excitation of multiple fluorophores in a single plane. Detection is via a CCD camera. The combination of optical sectioning and parallel image acquisition facilitates the rapid and efficient collection of data. This system can be used to view either fixed, fixed and cleared, or living specimens. Incubation, CO2 control and chamber perfusion are available. Available objectives include 5X (dry), 20X (water immersion) and 40X (water immersion). 6 laser lines: 405nm, 440nm, 488nm, 514nm, 561nm and 640nm are available.