Instruments
Laser Scanning Confocal Microscope I (FV1000 Olympus)
Microscope of choice for the following studies:
- Slides with fluorescent labels on cultured cells or tissue sections which require high resolution imaging
- Samples up to 100 um thicknesses
- Live imaging studies both short- and long-term including FRET, FRAP, FLIP, Photobleaching
- Multiple time point ("mark and find" studies) and studies requiring image tiling
- All fluorophores within the visible range with excitations beginning at 405nm can be imaged with this spectral laser confocal system. See below for laser lines.
Summary of Instrument Features
- Microscope: Olympus IX81 (INVERTED microscope configuration i.e. objectives below the sample)
- Objectives:
- Normal configuration: 4x (N.A. 0.16),, 10x (N.A. 0.40), 20x oil (N.A. 0.85), 40x oil (N.A. 1.30), 60x oil (N.A. 1.42), 100x oil (N.A. 1.40); Dry objectives available: 60x water .
- Dry objectives available: 20x dry, (N.A. 0.75), 40x dry (N.A. 0.60), 60x dry (N.A. 0.70)
- Water objectives available: 60x water (N.A. 1.20 (double check)
- DIC Capabilities: 20x, 40x, 60x, 100x
- Lasers: 405 diode laser, 440 diode laser, 488/515 argon laser, 559 diode laser, 635 diode laser
- Stage: Prior, Inc. motorized stage, includes tiling and multi-point mark and find memory function
- Stage Holders: Multiple stage holder formats are provided by the Core. Current stage holders will support: 1 x 3 slides, 2 x 3 slides, circular (all sizes) glass-bottom culture dishes, well plates with glass-coverslip bottoms
- Environmental Chamber: FULL STAGE INCUBATOR on the microscope, with CO2, temperature, and humidity regulation. Short- or long-term live imaging studies including FRET, FRAP, Multiple point timelapse imaging, etc. are possible
- New Binder Incubator in room: Cultures can be equilibrated for a number of hours before being transferred into the stage incubator with identical C02, temp. and humidity conditions.
- Service Contract Support: The FV1000 is under full service contract by Olympus.
- Opportunities to work directly with Olympus reps: The core organizes multiple opportunities for PIs and staff to talk or work directly with the Olympus rep when they are on-campus.
Conventional Microscope (Olympus VanoxAH-2)
Microscope of choice for the following study types:
- Studies needing brightfield microscopy
- Samples with strong fluorophore signal needing standard conventional fluorescence imaging
- Studies requiring "Live counting" image analysis
Summary of Instrument Features
- Microscope: Upright microscope configuration (i.e. objectives above the sample)
- Objectives: 4x, 10x, 20x, 40x, 100x
- Lamp source: Mercury Lamp
- Stage: Prior, Inc. Motorized Stage
- Rotatable slide holder: full rotation capabilities
- Camera: MicroFire Color Camera
- Image capture software: PictureFrame
- Image analysis software: StereoInvestigator (version 8.0)
- SI modules currently in Core:
- Optical Fractionator
- Confocal quantitation module
- Space Balls module
- SI modules currently in Core:
Laser Capture Microdissection Microscope (Zeiss Palmbeam)
Microscope of choice for the following study types:
* Capture of groups of phenotypically similar cells from fresh frozen, fixed then frozen, FFPE tissue onto eppendorf tube caps for further analysis by DNA, RNA, and protein assays
* Capture of cultured cells for further expansion or downstream processing for DNA, RNA, and protein analyses
Summary of Instrument Features
* Microscope: Upright microscope configuration (i.e. objectives above sample)
* Objectives: 10x, 20x, 40x, 63x dry - long working distances (able to see through back of slides with good resolution)
* LED fluorescence lightsource: Xcite Lamp
* Capture laser: Pulsed UV laser (100 nanosecond pulses)
* Cap Holder: Holds two caps
* Sample holders:
a)_for 1x 3 slides – holds 3 slides at a time
b) Culture holders
* Cameras: two cameras:
a) CC1 color camera
b) mRM camera for fluorescence
{Note: Both cameras provide the ability to capture “before cut, after cut, and on-cap” images}
* Stage: motorized stage
High Definition Camera with Adjustable Photography Stand for Large Specimen Imaging
Imaging equipment of choice for:
- Studies requiring comparison of histologically processed sections to the original thick slice of tissue and original organ (i.e. comparative microscopy for gross organ, macro slices to paraffin-embedded histology samples).
Summary of Instrument Features
- Stand: Kodak adjustable photography stand
- Camera: SPOT Camera
- Software: Image Pro Plus
Image Analysis Capabilities
At the current time, the following image analysis software packages are offered through the Core:
- StereoInvestigator for quantitation of counts of cells using the Optical Fractionator method, version 8.0
- Image Pro Plus
- FV1000 Software - basic analysis of confocal images
Contact Us
Director:
Linda Callahan
Phone: 275-1317
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