Effective date: June 1, 2021 (revised January 21, 2025)
Applies to: RII Imaging Cores - Optical
Brief Description: Failure to cancel a scheduled reservation more than 24hrs before the reservation begins will lead to the customer being charged for the reserved time. If the available time happens to be used by a different customer, the original customer is only liable for the remainder. See policy PDF below for specifics.

The TPI 1000 is available at the Marley location. A vibratome is useful for sectioning fresh or fixed tissues into thicker slices, such as 100-200um in thickness. It features an adjustable sectioning window, a bath drain, stroke pause switch, and an anti-corrosion surface. It has a total vertical specimen stroke of 15mm.

Users will need to receive training before they can access the vibratome, as the vibrating razor blade has the potential to be very hazardous.

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)

We have powerful workstations available with selected software programs (commercial or open source) for the post-processing and image analysis of microscope images. The workstations are available for scheduling in iLab. If you have specific post-processing needs regarding microscope data collected on our instruments, we will usually address these as part of the training on the instrument used to gather the data. Please feel free to ask us for assistance.

Depending on your needs (and our current expertise), we may be able to assist you in building an image analysis workflow. We have experience in thresholding and segmentation type analysis, but more elaborate workflows may be beyond our skillset or the capabilities of the software that we have available. Features that can often be quantitated include object counting, geometrical properties (length, area, perimeter, shape factors), and volumes can be estimated using stereological techniques. If we build a workflow, there may be a labor charge for our time. 

Our thoughts on fluorescence quantitation - the software to quantitate fluorescence intensities is easy to find. Unfortunately microscopes are known to be subject to a number of physics, engineering, and optical variabilities that are difficult to control. This means that to correctly capture fluorescence data that can be accurately quantified, the user must be willing to be meticulous in their sample prep and to ensure that the instrument variables are carefully controlled for (as a start, see The 39 Steps: A Cautionary Tale of Quantitative 3-D Fluorescence Microscopy). Controlling the instrument to ensure accurate data is time consuming and non-trivial. It is our opinion that much of what passes as quantitative fluorescence in the published scientific literature is worthless.