Policies for Safe Use of Shared Research Equipment
- All users must make an appointment by phone (402-559-6659) or email to the facility manager.
- Use on each instrument will be staggered so that there will be sufficient time between users for proper disinfection and cleaning.
- Saran wrap with a clean rubber band will be placed on each eyepiece by core personnel, to be removed and replaced after each user leaves
- In addition, after each use, the microscope eyepieces and all touch surfaces will be disinfected with 70% ethanol and Kim wipes.
- Users should not come to the UAMCF or be on campus if ill.
- All users entering the facility must don masks or face coverings over mouth and nose.
- All users entering will be immediately required to wash hands with warm water and soap before proceeding to the microscope room. Taped spaces on the floor will indicate where to stand for washing up.
- After washing hands, all users with then don gloves and safety goggles for the remainder of their time in the facility. When use of the microscope is completed, the users will deposit the reusable safety goggles in ready buckets with disinfecting solution.
- Only a single user per lab will be permitted per microscope.
- Use “after hours” (evenings/weekends) will be permitted but limited to ensure proper disinfection between users.
- Hours of operation are typically 8 am to 4 pm (by phone and email appointment only, no drop-ins) with opportunities to book time in advance from 4 pm to 7 pm Tuesdays and Thursdays.
- For any special issues or concerns, feel free to contact us directly.
- All users must have a cost center number entered into the system prior to each period of use. This is required for automated billing.
- Minimal charge will be 30 minutes with extended use charged to the nearest 1/4 hour. Billing statements are issued monthly.
- Individuals must be trained, accrue 20 hours of assisted image acquisitions, and receive permission from core staff before unassisted (after regular business hours and weekends) will be permitted.
- If you cancel with less than 24-hours’ notice, you will be charged a fee of $30 to recover instrument set-up costs.
- If you arrive more than 15 minutes after your planned arrival, your first hour will be charged from the time scheduled, not the time of arrival. If you are more than 30 minutes late during high use periods, your time slot may be assigned to another user.
- If you overbook time (i.e., book three hours and use one hour), you will be charged for the full use period unless another user is available on short notice.
Super Resolution Imaging (ELYRA PS.1) Policies
Structured Illumination Microscopy (SIM), Photoactivated Localization Microscopy (PALM), and Stochastic Optical Reconstruction Microscopy (STORM) are specialized techniques using delicate instrumentation requiring extended technical support from AMCF core staff. PALM and STORM require additional specialized sample preparations (see AMCF Imaging Resources).
- SIM imaging can be conducted using traditional blue, green, short- and long-red fluorophores that are bright.
- Users may bring SIM samples to core facility personnel for acquisition of images and basic processing.
- Only core personnel will acquire images and process them (including dual/multi-channel alignments). Users will be able to view and choose the regions of interest that they wish to acquire. The AMCF will consider independent usage after extensive training.
- The core facility will provide basic processing and channel alignment and will allow users to choose how they would like data rendered and saved. Among these options are TIF images of a composite 3D SIM, individual TIFs of various z-sections, altered angles, planes, etc. as TIF images, 3D movies, etc. Please note: these image processing manipulations are complex and take time, yet they are an essential part of the SIM analysis thus are part of the core service (and charge).
- First-time users may contact the core director for initial consultation/advise prior to using the facility. For each laboratory, this consultation will be free for the initial hour, but will be subject to time and availability of the director.
PALM, STORM Imaging (Currently in Development)
- The Elyra PS.1 is equipped for PALM, STORM imaging which requires the use of specialized photo-activated/photo-converted/blinking fluorophores and imaging buffers, see AMCF Imaging Resources.
- The AMCF will host a PALM and STORM imaging workshop with imaging specialists from Zeiss. This workshop will develop new SOPs and methodologies for researchers interested in super resolution imaging using PALM or STORM.
- PALM, STORM workshop: 07/2022 (exact dates pending)
- Research teams will bring fully prepared samples to their designated time slot during the super resolution imaging workshop
- Sample details and imaging goals must be submitted to the AMCF at least 2 weeks prior to the workshop
- Researchers may contact the AMCF Director to participate in the upcoming super resolution workshop.
In select circumstances, where additional expertise is required and goes beyond an additional consultation, if requested the director may agree to serve as a collaborator for a given project. This is subject to the discretion, time and availability of the director.
Acknowledging the AMCF Facility and Personnel
Services and equipment in the AMCF are provided with support from multiple funding agencies and MUST be appropriately cited for sustained operation of these shared resources. AMCF users are obligated to fully acknowledge the facility and its funding sources in formal publications and presentations containing data generated in the facility. We also appreciate receiving reprints or PDFs of such publications to use in our annual reports to internal and external funding agencies.
"This publication uses the following equipment, a [INSERT EQUIPMENT HERE*] housed in the University of Nebraska Medical Center Advanced Microscopy Core Facility (UNMC AMCF) supported by National Institutes of Health (National Institute for General Medical Science) INBRE P20 GM103427, Nebraska Center for Cellular Signaling COBRE P30 GM106397, NIH (National Cancer Institute) Fred & Pamela Buffett Cancer Center support grant P30 CA036727, the Nebraska Research Initiative and UNMC Office of the Vice Chancellor for Research. This content is solely the responsibility of the authors and does not necessarily represent the official views of the National Institutes of Health."
*Include all equipment used:
- Zeiss 710 CLSM (NIH S10 RR027301)
- Zeiss 800 CLSM with Airyscan (NIH P30 GM106397)
- Zeiss Cell Discoverer 7 live cell imaging system (NIH P30 GM106397)
- Zeiss Axioscan 7 whole slide scanner (NIH P20 GM103427)
- UltraMicroscope II Light Sheet Microscope (NIH S10 OD030486)
- HALO-equipped workstation (NIH P20 GM103427)
- IMARIS-equipped workstation (NIH S10 OD030486)
"We additionally thank [INSERT APPLICABLE NAMES HERE**] of the UNMC AMCF for their assistance."
**Janice A. Taylor, B.S., James R. Talaska, B.S., Heather Jensen-Smith, Ph.D.