Location Abbott 129

1. Leica SP2 AOBS LSM

Advanced confocal system with wide range of laser excitation lines (now we can do DAPI and Texas Red) and digital definition of wavelengths captured per detector channel (total of 3). Spectral scanning and separation of similar colored fluorophores possible (e.g., GFP vs FITC or YFP).

Microscope:

DM IRE2 HCB Fluo TCS /1-B-UV Inverted microscope stand DM IRE2 for transmitted light and fluorescence. Conventional fluorescence filter blocks:

* I3 (blue excitation BP 450 -490; long pass emission LP 515) typical for FITC or Cy2 etc.
* N2.1 (green excitation BP 515 - 560; long pass emission LP 590) typical for TRITC or Cy3 etc.
* Motorized coarse and fine focusing and 6-position objective turret.
* Objectives:
o 10x / NA 0.4 dry,
o 20x /0.7 dry, 40x / 1.2-0.75 oil,
o 63x / 1.4-0.6 UV oil, 63x / 1.3 glycerol,
o 100x /1.40-0.7 oil.
* DIC polarizer/analyzer plus prisms appropriate for each objective.
* Transmitted light detector for recording bright field images.
* Galvo stage with fast z-axis capability, and
* TMC anti-vibration table.

Scanner: Scan detector equipped with an acousto-optical beam splitter (AOBS) to select/introduce most excitation laser lines (9 available), eliminating the need for main dichroic mirrors. Three PMT detectors with 8 or 12-bit output; simultaneous operation, digital/graphic definition of emission wavelength content per channel. Spectral scanning / display and linear un-mixing: the ability to separate largely overlapping spectra.

The following laser lines (mW power) are available: 405 (25), 458 (5), 476 (5), 488 (20), 496, 514 (20), 543 (1.2), 594 (orange HeNe), 633 (10) nm, fiber coupled, and controlled by AOTFs. Scanner with 2000-Hz line scans, frame scan rate of 40 fps at 512x32 pixels, scan resolution up to 4096x4096 pixels, zoom to 32x, scan rotation; interactive control panel with digital potentiometers.

Control: LCS Leica Confocal Software with fully operator configurable user interface. Context sensitive online help system. Multitasking high performance Windows XPpro Xeon Workstation (HP X4000) with 2.4GHz CPU, 2048MB RAM, two 21"CRT Monitors, 60 GB SCSI hard disc, 3.5" floppy, 16x DVD ROM, CD writer 16x/10x/40x. Software includes LCS 3D Volume rendering, 3D animations, 3D filter, Stereo images and animations, Average projection, Maximum point projection, SFP; LCS Multicolor Co-localization of 2channel and 3channel recordings. Cytofluorograms in 2D and 3D display. Histogram-segmentation and masking.

2. Olympus Live-cell Automataed Multi-parameter Digital Imaging Microscope

Allows for automated acquisition of high resolution image data using multiple excitation and emission wavelengths over time and space (Z axis); fluorescence and DIC.

Microscope: Inverted Olympus IX81 side port stand, Tilting binocular, Motorized DIC/Phase Condenser, U Plan Fluorite 10X Objective, NA 0.30, LWD Plan FL 20X Objective, NA 0.40 with Correction Cap for 1.1mm Thick Plastic, CAP-G0.5 Correction Cap for Glass Vessels, 40X oil NA 1.35, TIRFM-SP Plan APO 60X Oil objective; NA 1.45, U Plan APO 60X Water objective, NA 1.20, Motorized 6 position fluorescence mirror cube turret; mirror cubes: DAPI, CFP, C/YFP FRET, YFP, red, Cy5, Fura-2. High power 100W Hg arc lamp. Integral Z/focus motorization, 10nm step resolution. High resolution (1360x1036, 6.45 mm pixel), high quantum efficiency Retiga EXi digital chilled CCD camera, firewire interface, 8 or 12-bit output. Frame rates are 10 and 5 fps at full resolution (8 vs 12 bits, respectively), to 120 fps for 8-bit, 4x4-binned, 10x10 pixel subregions; exposures from 40 microsec to 15 min. Colors filters are now installed on the Retiga camera to capture color images.

Control: Intel P4 (2GHz) Windows XPpro CPU with CDRW, 80GB hard disk, running MetaMorph and MetaFluor software. Ten-position Sutter excitation and emission filter changers, Fura-2 exciters, C/YFP FRET emission filters; neutral density filters.

3. Fluoview laser scanning confocal microscope

A user-friendly, high-resolution system for confocal optical sectioning of fixed and living specimens and data analyses. The system features dual detectors for simultaneous data acquisition of green and red/far red fluorescence or DIC transmitted light images, automated optical sectioning, time lapsing, and device control (for physiological experiments).
3 ion lasers, 3 laser lines. 488nm Ar, 543, 633nm HeNe's; illumination by individual lines and 488+543 or 488+633 dual lines
dual scanning galvanometer mirrors (1.5 msec/line scan rate)
neutral density selection of 100, 50, 20, 6% laser light
5 confocal pinhole apertures
barrier filter selection for narrow or long-pass green, red, and far red (Cy5), or none for reflectance and transmitted light modes
two photomultiplier detectors, 12-bit, image resolution to 1024x1024 pixels
digital zoom 1-10X
z- stepping motor, 0.1 µm resolution (will accept piezo steppers)
scan control for x-y, xyt, xyz, xyzt, xz, and xt modes; region-of-interest scan
Tiempo physiology software (real-time intensity plots, ratio calculation, calibration, trigger in/out)
Extensive analysis capability (distance, area, intensity, multichannel, volume reconstruction, animation, tiling, math/logical, masking, digital filtering, color encoding/overlay)
Olympus IX70 (inverted) microscope with conventional fluorescence ( 100W Hg) and DIC optics
Wide GFP and "rhodamine" filter sets
100x ,60x,40x,20x,10x
PIII-450MHz PC with 512MB RAM, 68GB storage, CD, CDRW, floppy, and zip drives; ethernet and server mode.

4. Zeiss Axiovert 135

Link to old SBRI web page:

http://digital.bsd.uchicago.edu/merged%20sbri.html

Contact:

Christine Labno, PhD
Technical Director
Tel. 773.834.9040
email: ccase@midway.uchicago.edu