Imaging Cores - Optical

RRID:SCR_023355

Microscope training - specific to each instrument

Each instrument in our core facility requires specific training by one of our core managers before a user can be allowed to operate it by themselves. The exact length of training varies due to the complexity of the instrument and the previous experience of the new user. Training is charged at the the instrument's routine hourly rate plus a per hour staff labor fee (training rates for all of our instruments can be found in iLab). The core manager makes the determination as to when the user is ready to operate an instrument by themselves.

Training is often 2-3 sessions of 1.5 hours each. Because our rooms are small, we limit the number of trainees to a maximum of two individuals from the same lab. The first session is typically instructional, with the manager explaining principles of the optics, electronics, software, and data handling. The core sometimes uses a teaching sample for the first session. The remaining sessions are hands-on with the user's sample and the user learning to operate the microscope. We teach the trainee how to get the most of the instrument and often make suggestions about sample prep, alternate techniques, controls, etc.

For many of our instruments we have written resources (available online as PDFs) and we are working on developing D2L-based training to replace the instructional sessions for most of our instruments.

Please contact us if you have any questions.

Service

Project Consultation

Getting the most out of our microscopes requires careful experimental design and sample preparation. We are available to discuss your research needs, review your plans, suggest changes that could improve your images, and steer you towards an instrument that is best suited to your needs (even if it is in another core). Please contact us to talk about your research needs.

Some of our microscopes can be used for BSL2 research. DO NOT SHOW UP WITH A BSL2 SAMPLE WITHOUT FIRST CLEARING IT WITH THE ON-SITE CORE MANAGER!

If your questions require us to do additional in-depth research, we are happy to do that. There may need to be a labor charge for this extra work, but that can be discussed up front.

Service

Zeiss LSM880 post-processing computer workstation

This workstation is only available to users already trained on the Zeiss LSM880 microscopes. The PC is a Hewlett-Packard Z6 workstation with an 8 core Intel Xeon CPU, 64GB of RAM, 12TB of data storage (RAID 10), running Windows 10 (64 bit) with a licensed version of the Zeiss ZEN software for data processing (stitching, spectral unmixing, etc.) and FIJI (ImageJ).

Equipment
Location

Pelco Biowave Pro Microwave Tissue Processor

Programmable microwave tissue processor, with continuous power settings from 100-750 Watts, load cooling to maintain consistent temperatures, with built-in fume extraction and vacuum.

The microwave is useful for:

  • improved tissue fixation
  • improved retention of fluorescence after fixation
  • for more rapid tissue processing for immunocytochemistry
  • electron microscopy
  • decalcification.
Image
Microwave
Equipment
Location

Zeiss Axiozoom 16 Fluorescent Stereo Microscope

The Zeiss Axiozoom 16 fluorescent stereo microscope is equipped with a Hamamatsu Flash 4.0 camera. The HP computer is equipped with Zen Blue software for combining the grayscale images from each channel into color overlays. The fluorescent filters include UV (DAPI), Blue (GFP, FITC, Alexa488) and Green (RFP, Alexa 555). The magnification ranges from 7-112x (we now have a 2x front lens that can be mounted on request, allowing for a magnification range of 14-224x). There is also a fiber optic light to enable brightfield images.
Image
Zeiss_Axiozoom16
Equipment
Location

Zeiss LSM880 NLO Upright Multiphoton/Confocal Microscope with Fast Airyscan

The Zeiss LSM 880 NLO upright multiphoton/confocal microscope includes a 34-channel detector for spectral information and additional non-descanned GAsP detectors. The system includes 7 visible spectrum laser lines (405, 458, 488, 514, 561, 633 nm), plus an additional software-tunable Spectra Physics Mai Tai HP DeepSee laser (690-1040 nm). The system includes software modules for controlled region-of-interest scanning, deconvolution, physiology experiments, performing FRAP & FCS and control of the motorized stage to allow stitching together images from multiple fields of view into one larger image (montage). The software also enables visiting multiple sites during a time lapse acquisition, and the instrument includes an environmental chamber suitable for live cell imaging at a variety of temperatures. Wet lab space is available to users of the facility in an adjacent room, including cell culture incubators and a biosafety hood.

NOTE: BSL2 samples must be discussed with the on-site core manager BEFORE they arrive in the facility. Live animals - invertebrates only!

The Fast Airyscan allows for resolution improvement up to 1.9x of typical confocal resolution and the fast mode sacrifices a bit of resolution for speed (useful for live cell imaging). The scanhead includes a 34-channel detector for spectral information.

Commonly used lenses on the Upright Zeiss
LSM800 NLO confocal microscope

Numerical
aperture

Working
distance (mm)

type

20x plan apo

0.8

0.55

dry

40x plan apo

1.3

0.17

oil

20x plan apo dippable

1.0

2.4

water

40x plan dippable

0.8

2.1

water

Image
LSM880 NLO
Equipment
Location

Zeiss LSM880 Inverted Confocal Microscope

The Zeiss LSM 880 inverted confocal microscope includes a 34-channel detector for spectral information, and 7 Laser lines (405, 458, 488, 514, 561, 594, 633nm). The system includes additional software modules for performing FRAP and control for the motorized stage to allow stitching together images from multiple fields of view into one larger image (montage), as well as visiting multiple sites during a time lapse acquisition. The instrument includes an environmental chamber suitable for live cell imaging at a variety of temperatures, humidity and CO2 capabilities. The facility has wet lab space, cell culture incubators, and a biosafety hood in an adjacent room.

NOTE: BSL2 samples must be discussed with the on-site core manager BEFORE they arrive in the facility.

Objective lenses (with matching DIC prisms) include:

  • Plan-Apochromat 10x/0.45 (WD=2.0mm)
  • Plan-Apochromat 20x/0.8 (WD=0.55mm)
  • Plan-Apochromat 40x/1.3 Oil
  • Plan-Apochromat 63x/1.4 Oil
  • Plan-Apochromat 40x/0.95 dry CORR (not typically on the microscope, available upon request)
Image
zeiss lsm880
Equipment
Location