This is a multi-function, motorized inverted microscope that includes a brightfield color camera and a 14bit greyscale fluorescence camera. The addition of the Apotome III attachment allows this microscope to give crisp "confocal-like" z-stack images of thinner samples. Thinner means out to approximately 20um in thickness, making this microscope suitable for samples such as: cells on a coverslip, paraffin and standard cryostat sections.

How does this microscope create "confocal-like" images? - The Apotome III projects a grid structure into the focal plane of the specimen, then moves it into different positions using a scanning mechanism. At each grid position, the Apotome III automatically acquires a digital image. The system processes all images into one optical section with improved contrast and increased resolution using a patented algorithm. The resulting image is free from grid structures. (SOURCE: Zeiss) Deconvolution software is available to further improve the image quality of the Apotome images.

Features:

• AI sample finder - using a unique contrasting method, the microscope can find multiple wells in a plate, unstained sections on a slide, or locate the edges of a coverslip . Very useful for setting things up to scan multiple areas or large area scanning of tissue that's otherwise difficult to see by eye.
• Colibri 7 LED illuminator allows for 7 specific excitation wavelengths (including a near-IR wavelength). With just two multi-band filter sets, switching between channels is significantly faster than microscopes using filter cubes, making tile scanning in fluorescence much faster. Due to spectral overlap issues, we should be able to capture up to 5 channels in regular widefield fluorescence and 4 channels with the Apotome III (which does not work with near-IR wavelengths). To see how your dyes might work with this system, see our configuration at FPbase.
• Capable of color brightfield imaging (e.g., histology stained slides), Differential Interference Contrast (DIC, e.g., unstained cells), simple Polarized light imaging (crossed linear polarizers, e.g., Picrosirious red staining), and multiple channels of fluorescence as well as combinations of any of these imaging modes.

Specifications

• Objective lenses: 5x/0.16NA dry, 10x/0.45NA dry, 20x/0.8NA dry, 40x/1.3NA oil, 63x/1.4NA oil (a 40x/0.95NA dry objective with coverslip correction is available upon advance request)
• Colibri 7 excitation wavelengths: UV 385/30nm, V 423/44nm, B 469/38nm, G 555/30nm, Y 590/27nm, R 631/33nm, FR 735/40nm
• Zeiss 489112-9110-000 (suitable for fluorescent dyes like DAPI, FITC, DsRed, CY5, and CY7) and 489091-9110-000 (suitable for fluorescent dyes like CFP, YFP, and mCherry) multiband filter sets

Post-processing of image data includes:

• Exporting from propriety formats (e.g., Zeiss CZI or Leica LIF) to TIF images
• stitching of tile scans (montage creation)
• spectral un-mixing of data collected using the Quasar detector on either of the Zeiss LSM880 microscopes
• Airyscan post processing (Zeiss LSM800 confocal/multiphoton - due to software licensing issues, this is only available on the microscope computer)
• SIM post-processing and channel alignment (Zeiss Elyra S.1)
• Deconvolution of Apotome data (Zeiss Axio Observer 7 with Apotome III Microscope)

If users need these post-processing techniques, they are typically covered in the user's training on the specific instrument. The workstations are available for scheduling on iLab to approved users. If you need training or assistance, please feel free to contact us.

• Image Analysis computer workstation (LSN)
• Zeiss SIM post-processing computer workstation (LSN)
• Zeiss LSM880 post-processing computer workstation (Marley)

There are times when users need quality microscope data immediately, or they may only need a few example images to compliment other scientific data for a publication or a grant submission. Our experienced core managers are available to operate the microscopes on your behalf. The charges for assisted use are calculated at the routine hourly usage rate plus an hourly staff labor charge (rates are available in iLab).

Sample preparation strongly affects the quality of the images that can be captured with a microscope. If a user brings us a bad sample, we will give the user the best images that are possible with that sample. Please consult with us ahead of time to ensure a good outcome.

While we are very familiar with our instruments, occasionally we are asked to perform an imaging method that we have not done before. This may require additional research on our part, and perhaps additional training from the vendor before we can successfully perform the new method. Depending on the amount of time involved, we may need to discuss an additional staff labor fee.

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

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 in 2023 we will be working on developing D2L-based training to replace the instructional sessions for most of our instruments.

Please contact us if you have any questions.

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.