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College of Pharmacy

Microscopy Core

Instrumentation

The Microscopy Core hosts a wide variety of biological imaging technologies for researchers.

LEICA  ASMDW

Leica AS MDW Live Cell Imaging System

Our Leica AS MDW system was developed for live cell fluorescence imaging. Features include:

Xe monochromator for narrow excitation band
Motorized stage for multiple x,y positions per time point
Specimen chamber: temperature, 5% CO2 atmosphere

The AS MDW was developed for live cell fluorescence imaging. The system uses the Leica DM IRE2 inverted stand with motorized objective turret, z positioning, and fluorescence filter changing. In addition to a mercury lamp it has a Xenon lamp in a monochromator. This latter source is more stable over time and causes less phototoxicity in comparison to the mercury lamp.

The AS MDW also has a motorized stage so that multiple fields can be imaged, over time, in the same experiment, increasing the number of cells analyzed. There is a humidified chamber which fits on the stage and can be perfused with custom mixed gases, such as 5% CO2 in air, to maintain the proper pH of bicarbonate-buffered media.

The system is equipped with N PLAN L 40x/0.55 CORR, HCX PL APO 63x/1.30 GLYCEROL and HCX PL APO 100x/1.4 OIL objectives. Differential interference contrast optics provide contrast in bright field images. The 63x objective is optimized for 37°C and is on a Piezo mount for 3ms focus changes during z stack acquisition. On the bottom port of the stand is a Photometrics Coolsnap HQ camera. Multiple image planes can be acquired at each time point and the resulting data can be processed by 3D image deconvolution.

FLUORESCENCE IMAGING

2 Leica DM IRE2 Microsystems

We have two Leica DM IRE2 systems, one within an environmental chamber for live cell imaging. Features include:

— Environmental chamber
— 10x to 100x
— DAPI (blue) , FITC (green), TRITC (red), and Cy5 (far red) filter sets

There are two of these systems, one within an environmental chamber for live cell imaging. Each uses the Leica DM IRE2 inverted stand with motorized objective turret, z positioning, and fluorescence filter changing. The objectives on each include 10x/0.25n.a. Ph1, 40x/0.6n.a. Ph2 HCX PL FLUOTAR, 63x/1.4 HCX PL APO and 100x/1.4 HCX PL APO. The microscopes use phase contrast with the 10x and 40x objectives and differential interference contrast with the 63x objective.

100W mercury arc lamps provide excitatory light for fluorescence imaging and there are filter cubes for DAPI, GFP (FITC & Cy2), TRITC (Cy3 and Texas Red) and Cy5 (Alexafluor633). Both DM IRE2 microscopes have CCD cameras for fluorescence imaging, one a Leica DC350 FX and the other a Photometrics Coolsnap HQ. The microscopes and cameras are controlled with personal computers running Leica FW4000 software. The software enables the user to set up image acquisitions incorporating z-series, time series and multiple color channels. The FW4000 software also performs basic analysis and image processing functions.

Live Cell Imaging

3 Carl Zeiss Axiovert 200 Microscopes

We have three identically configured Zeiss Axiovert microscopes on air tables for routine phase contrast and color imaging of live and stained specimens.

Cell migration
— Proliferation
— Apoptosis
— Mitosis
— Cell lineage tracking

There are three identically configured Zeiss Axiovert microscopes on air tables. Each is equipped with 10x, 20x and 32x objectives, phase contrast optics, a Zeiss AxioCam MRc color camera and a PC interface running Zeiss Axiovision. These are used for routine phase contrast and color imaging of live and stained specimens.

Olympus  IX81

Olympus IX81 Fluorescence Microscope

The Olympus IX81 is an inverted system microscope built for live cell imaging. Features include:

— Heated stage insert
— Filter wheel for short-term live cell imaging of rapid processes

The Olympus IX81 has an inverted motorized stand controlled by the IX2-UCB control box. Fluorescence illumination is from a Sutter Instruments Lambda LS Xenon lamp with liquid light guide. Fluorescence filter changing is done by the Sutter Instruments Lambda 10-3 filter changer. There are filter cubes for DAPI, FITC, and TRITC. For more rapid filter changing for live cell imaging there is a filter wheel containing emission filters for DAPI, FITC, TRITC and Cy5. Also for live cell imaging there is a Bioptechs Delta T5 culture dish temperature controller. The objectives include a UPlanFL N 10x/0.30 Ph1, a LUCPlan FL N 40x/0.60 Ph2 with correction collar for long working distance, a UApo/340 40x/1.35 oil iris which transmits UV light for 340/380 ratio imaging, a PlanApo N 60x/1.42 oil and a UPlanSApo 100x/1.40.

CARL ZEISS LSM700

Carl Zeiss LSM700 Confocal Microscope

Our LSM700 laser scanning microscope is a member of the seventh generation of confocal instruments from Carl Zeiss. Features include:

Confocal point scanning
3D reconstruction
— Multi-field time lapse
— Ratiometric concentration determination
— Tile and stitch
— FRET (Förster resonance energy transfer)
— FRAP (fluorescence recovery after photobleaching)

The LSM700 has a  scan head  with two photomultiplier tube  (PMT) detectors on an inverted Axio Observer Z1 stand. There are four solid state lasers producing lines at 405nm, 635nm, 555nm and 488nm. A transmitted light PMT collects transmitted light images using differential. A secondary dichroic beamsplitter facilitates acquiring lambda scans to be used for linear unmixing. 

The entire microscope stage and objective turret are enclosed in an environmental chamber to have stable temperature and CO2 control for live cell imaging. There is a motorized stage for acquiring multiple fields in separate wells of a multiwell plate at each time point in time lapse data acquisition. 

zeiss palm system

ZEISS PALM System for Laser Microdissection

ZEISS PALM system for laser microdissection combines powerful imaging capabilities with robust and operationally simple microdissection technology. 

Applicable to cryosections, FFPE (formalin-fixed and paraffin embedded) tissue, native tissue like fresh plants, live cells and chromosomes - without contamination
— Suitable for laser microdissection and analysis for DNA, RNA and protein isolation, whether from archive material or live cells
— Works with standard glass slides

ZEISS PALM system for laser microdissection combines powerful imaging capabilities (bright field, phase contrast, multi-channel fluorescence) with robust and operationally simple microdissection technology. 

PALM MicroBeam uses a focused laser beam to cut out and isolate the selected specimen without contact. The patented laser catapult isolates the target area fast and uncontaminated. It allows to obtain the homogenous analysis material necessary for meaningful scientific results.  The short laser pulse minimizes time heat transfer to adjacent areas, allowing isolation of vital life cells.

Because analyses of gene expression patterns rely on exactly-separated analytical material, unwanted cells may alter your results and conceal the signals of the relevant cells. PALM MicroBeam prevents this by allowing you to define cells and tissue regions precisely, ensuring your results are exact and reproducible.

 


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