Confocal Microscopes


Zeiss LSM 800 Laser Scanning Confocal Microscope (A/Z W5)

The Zeiss LSM 510-Meta microscope located in A/Z W5 was recently upgraded to an LSM 800. The new system includes solid state lasers at 405 nm, 488 nm, 561 nm and 642 nm. The objectives currently installed vary from 5x to 63x magnifications. Other features include a motorized 3D positioning stage, X-Cite LED transmitted light with filters for blue, green and red epifluorescence, a Zeiss Axiocam color camera, a Definite Focus module for autofocusing correction, and three dedicated collection channels including one GaAsP detector. The microscope includes Zen Blue acquisition and processing software.

Cost

$40 per hour flat rate.

Access/Sign-up Process

To schedule time on this microscope, please contact the facility supervisor, Connie King, Research Associate, Tel: 1-6608.


Zeiss LSM 510-Meta Laser Scanning Confocal Microscope (Microbiology, B318)

Cost

The user fees for this instrument are as follows:

  • Hourly use for trained users: $40/h

  • Fee-for-service: $25/h (in addition to hourly use fee)

  • Computer log-in will keep track of use time and billing will be done bi-monthly by electronic billing of pre-registered account numbers.

  • Costs for users external to CSU will be at $80/h plus $50/h fee-for-service.

Access/Sign-up Process

Access is limited to individuals that have passed a basic class in operation and care of the system. To schedule time on this microscope, please contact the facility supervisor, Dr. Mercedes Gonzalez-Juarrero, Associate Professor, Tel: 1-7306.


Olympus IX81 Inverted Confocal Laser Scanning Microscope (Research Innovation Center, Foothills Campus)

The Olympus IX81 Inverted Confocal Laser Scanning Microscope is located in the Research Innovation Center, Foothills Campus has widefield fluorescence visualization, brightfield and differential interference contrast. Four channel spectral and filter-based confocal scanning includes a transmitted light channel for DIC. This feature provides sensitivity and flexibility for imaging with and without fluorescent fluorophores and will allow users to image up to four compatible fluorophores simultaneously.  Objectives on this system are 10X, 20X, 40X, and 100X. This instrument also has a second set of galvanometer mirrors allowing simultaneous photo-activation/stimulation/bleaching during imaging experiments. The second scan unit is a laser point scanner with the capacity to target defined regions using a Tornado (helical) scan. This feature allows activation and depletion of fluorescent probes for analysis of temporal and spatial activity in live samples. A spectral detector based PMT system allows for specific bandwidth selection in co-localization experiments plus spectral scanning and separation of overlapping fluorophores. The FV1000 also has a motorized, programmable stage that can be used for multipoint time-lapse and mosaic imaging. The four lasers with AOTF control allow six different wavelengths to be selected: 405 nm, 457 nm, 488 nm, 514 nm, 559 nm and 635 nm. Other features include multimode imaging in six dimensions including: XYZ, time, wavelength, mosaic scanning, and multipoint high content imaging.  Fluorescence Recovery after Photobleaching (FRAP) and Photoactivation (PA) is facilitated by the easy software wizard with any laser line. The system also includes co-localization software module for complete analysis including scattergrams and statistical data ready for easy export.

The FV1000 is an Olympus IX81 Inverted Microscope with widefield fluorescence visualization, brightfield and differential interference contrast. The system allows for four channel spectral and filter-based confocal scanning including transmitted light channel for DIC. This feature provides sensitivity and flexibility for imaging with and without fluorescent fluorophores and will allow users to image up to four compatible fluorophores simultaneously. We have 10X, 20X, 40X, and 100X objectives for use on this microscope.

A key feature of this instrument that was not available from other manufacturers is the incorporation of a second set of galvanometer mirrors for simultaneous photo-activation/stimulation/bleaching during imaging experiments. The second scan unit is a laser point scanner with the capacity to target defined regions using a Tornado (helical) scan. This feature allows activation and depletion of fluorescent probes for analysis of temporal and spatial activity in live samples.

This spectral detector based PMT system allows for specific bandwidth selection in colocalization experiments plus spectral scanning and separation of overlapping fluorophores. The FV1000 also has a motorized, programmable stage that can be used for multipoint timelapse and mosaic imaging. The six line laser combiner with AOTF control includes the following lines that will provide us the flexibility required to look at multiple fluorophores:

405                                                                 DAPI, Hoescht, Alexa 405

mArgon (457nm, 488nm, 514nm)              CFP, GFP, FITC, Alexa 488, YFP

559                                                                 TRITC, CY3, mCherry, DsRed

635                                                                 Cy5, Alexa 647

Other features that this model includes multimode imaging in six dimensions including: XYZ, time, wavelength, mosaic scanning, and multipoint high content imaging.The Fluorescence Recovery after Photobleaching (FRAP) and Photoactivation (PA) that is facilitated by the easy software wizard with any laser line. The system also includes colocalization software module for complete analysis including scattergrams and statistical data ready for easy export.

For access to this microscope, contact the facility supervisors, Prof. Brad Borlee: brad.borlee@colostate.edu and Grace Borlee: grace.borlee@colostate.edu.


Olympus IX83 Inverted Spinning Disk Confocal Microscope (MRB 224)

The Olympus IX83 Inverted Spinning Disk Confocal (SDC) Microscope was recently upgraded from an IX81 microscope frame to include a Zero Drift Compensation (ZDC) autofocusing unit. The microscope has a custom-built stage top environmental control chamber with CO2 control for live cell work, an X,Y, piezo Z stage for rapid image stack acquisition across many predetermined fields (4 D imaging), a CSU 22 head with quad dichroic and additional emission filter wheel to eliminate spectral crossover, four high power diode lasers (405 nm, 488 nm, 561 nm and 647 nm) with rapid (microsecond) switcher and a phasor holographic photobleaching/photoactivation/photoconversion system for intracellular molecular dynamic measurements. Phasor can also be used for ion uncaging and other applications. System has differential interference contrast (DIC) optics with 10, 20, 40, 60 and 100X objectives, built in correction for spherical aberration for all objectives, and a wide field Xenon light source.  A cascade II EMCCD camera (confocal imaging) and a Photometrics HQ camera (wide field imaging) are both integrated for image capture using Slidebook software.  Laser selection and power are software controlled.  System is connected to RAID terabyte storage. 

Data Storage

Users must bring their own drives for downloading image files as soon as the experiments are complete. Storage capacity for a longer time may be purchased on the Terabyte drive system.

Cost

Training: Cost for training is $250, which includes 5 hours of microscope access after training.

Short-term use: $20 per hour flat rate.

Long-term use: New users pay $20 per hour up to a maximum of $400 for the first month, $300 for the second month and $210 per month thereafter. The increased rates for the first two months reflect the additional supervisory time and maintenance/service required for new users. Costs decline as users become proficient in application of the many different items of ancillary equipment and program applications associated with the microscopes.

Trained users pay $20 per hour to a maximum of $210 per month for unlimited access.

Fee-for-service: These projects are negotiated on a project by project basis, but generally cost $60 per hour in addition to the microscope user fees.

Scheduling

To schedule time on the SDC, please contact the facility supervisor, Dr. Barbara Bernstein, Senior Research Scientist, Tel: 1-0430.

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