Microscopy

The Microscopy Core Facility provides researchers with the ability to visualize a variety of samples from single fibril and intracellular structures to very thick sections from organ tissues. This is accomplished using a variety of imaging technologies, including: Widefield, Light sheet (DLS), Confocal (LSM), Multi-photon, Total Internal Reflectance Fluorescence (TIRF), Super-Resolution Stimulated Emission Depletion (STED) microscopy, as well as equipment for cell and tissue laser dissection microscopy and live cell imaging and image analysis. The Core Facility also allows users to conduct biomechanical studies complemented by the Atomic Force Microscopy (AFM) including force spectroscopy experiments and imaging. The Microscopy Core Facility provides access to several workstations with software packages for image processing, analysis, and 3D image reconstruction. The Core Facility director offers instrument training, imaging and sample preparation assistance, as well as consultation for both on- and off-campus investigators of all levels of experience and from any discipline.

Contact Information:
Sergey Pryshchep, Ph.D.
Microscopy & Cellular Imaging Core Director
518-276-3456
pryshs@rpi.edu

Access to the Microscopy Suite and Equipment

Access to all equipment must be approved by the core director. Approved users will have their ID cards activated to access the microscopy suite. Center users may request keys to the equipment rooms (limit one key per laboratory for each equipment room). All other users will need to request temporary access from the front desk.
Each individual user has a unique login name. Password sharing is prohibited. Doing so will lead to suspension of user’s access to the facility.

Training

All users must receive proper training before being permitted access to the equipment. Training is provided by the core director. The length of the training varies with the trainee’s prior experience on the equipment, but a typical training session will last about three hours. Minimum training time is one hour.

Scheduling

Advance reservation using the online sign-up is required. The billing is based on the length of occupation time, which is calculated based on the online sign-up records (minimum) and the equipment computer log records. Make sure to logout after each session.

Cancellation within 48 hours prior to the experiment will be subjected to a $20 cancellation fee.

Cleaning Up

After each session, users are required to remove oil from the objectives using the lens paper and the Sparkle solution (purple color) as demonstrated during the training. Do not rub the lens.

Users are responsible for keeping the equipment areas clean and for removing their data from the hard disk of the computer immediately after each session. Long term storage of large quantity of data on the equipment computer is prohibited.

Log Book

The logbook must be used to record the actual start and end time for each session and any problems encountered during the session.

Problems, Questions and Suggestions

Users are required to report to the core director any problems that occur on the equipment during the session immediately. If the core director is unavailable (e.g. after hours use), please e-mail the core director and leave a note for subsequent users if the problem affects the usability of the instrument. Questions and suggestions are always welcome.

Billing

All users are responsible for providing accurate and updated billing information. Users with multiple billing accounts should specify the account number to be charged upon sign-up. Both internal and external users will be billed on a monthly basis.

Core User Grants

Unfunded investigators who need to use the facility may apply for a core user grant. Eligibility will be determined by the Center Director, Dr. Deepak Vashishth.

Technology

Example of data produced by the Leica TCS STED Microscope
The Leica TCS SP8 3X STED offer flexible system solution focused on live cell as well as fixed sample super resolution imaging. The Leica 3D STED instrument achieves resolution below the diffraction limit (50-200nm) in both the lateral, and axial directions.
Example of data produced by the Multiphoton Microscope - Zeiss LSM
Ideal for imaging very thick fluorescently labeled specimens with less photobleaching and attenuation than conventional confocal. Many live specimens can be imaged without causing damage.
Example of image produced by the Zeiss LSM 510 META
This LSM510 confocal system is equipped with multiple laser lines ranging from 405nm to 633nm. The system uses a Zeiss inverted microscope stand. It can collect bright field and DIC as well as conventional and confocal fluorescence images. This microscope has the ability to measure in 3 dimensions, with software to automatically handle a series of optical sections in the Z dimension.
Image of the instrument and the type of results produced
The MFP-3D AFM provides high resolution images, exact force and stiffness measurement in both air and liquid. The MFP-3D AFM is integrated with an inverted optical microscope, supporting optical microscopy techniques including fluorescence microscopy.
Example of data produced by optical tweezers
Optical tweezers are capable of measuring picoNewton forces involved in single molecule dynamics and interactions (e.g. proteins & nucleic acids), while the laser microdissection setup can isolate tissue, single cells, or even submolecular structures (e.g. chromosomes); isolated material can then be prepared for genomic or proteomic analysis.
Data produced by the Olympus IX81 TIRF Microscope
Total internal reflection fluorescence (TIRF) microscopy is ideal for visualizing structures in the immediate vicinity (~100nm) of the coverslip including cell membrane structure and dynamics, and for resolving single molecules (e.g. DNA, cytoskeletal motility assays).
Leica CM150 Cryostat
The CM1520 from Leica is a cryostat for high quality sectioning of frozen tissues. The displays and instrument controls are easy to operate. This instrument is used to rapidly section frozen tissues such as brain, spleen, kidney, artery wall in thicknesses between 5 – 300 µm. All tissue samples are frozen with O.C.T. embedding medium to provide rigidity and support for sectioning.
Autosamdri®-815, Series A
Microelecromechanical systems, TEM and SEM sample. Process up to 5 x 10mm square die or 5 x 1" diameter wafers with every run.

Required Acknowledgement and Authorship

Please acknowledge the CBIS Core Facilities in all publications and grant applications where our equipment and/or personnel have facilitated the work. These acknowledgements are very important because documenting our contributions helps to ensure that the resources of the Core Facilities are sustainable.

  • Equipment: If you used Core Facility equipment, please note this in the Materials and Methods. e.g., Thermogravimetric analysis was carried out using a TA Instruments TGA-Q50 (Rensselaer CBIS Analytical Biochemistry Core Facility).
  • Personnel: Please consider including CBIS personnel as co-authors on your publications when they have made a significant intellectual contribution to the research. Include CBIS Core Facility directors or staff as co-PI or co-investigators in grant applications when they provide a significant contribution to the grant proposal and scientific/intellectual leadership for the proposed work. Please follow these guidelines: ABRF Recommended Guidelines for Authorship on Manuscripts. Also, our Core Facility personnel always appreciate when they are mentioned in the Acknowledgements section of publications.
  • Required Funding Authorization Form: Rensselaer researchers must fill out the CBIS Cores Authorization Form (PDF) to use the CBIS Core Facilities.
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