Perelman School of Medicine at the University of Pennsylvania

Electron Microscopy Resource Lab

Beckman Foundation
Electron Microscopy Resource Lab

Training & Services

All requests for services are done on the iLab site. Please Click Here to redirect to iLab website and choose the service and send your request. 

Services Offered at the Cryo-EM Screening Lab:

Negative stain/Pay-Per-Service (Internal only):

Imaging of biochemically pure viruses, proteins, nucleic acids, or liposomal complexes. All the supplies required for conducting the experiments are provided by the facility at an appropriate cost. However, users can bring their supplies if preferred.

Negative stain Imaging service on T12 (Internal Users only)

Service includes following

T12 Microscope time

3 hrs

Staff time

3 hrs

Negative stain solution

2% Uranyl Acetate in DI (pH ~6.0)

Blotting paper

As needed (Whatman filter paper)

Thin carbon 300 mesh copper grids

3

No of trials

3

Available Buffer for dilution

20mM Tris pH 8.0, 150mM NaCl

The user brings a report on the quality and quantity of the sample and a buffer for dilution.

Here, up to 3 hrs of T12 time and staff time are covered. A total of up to 3 different conditions and trials are executed. The optimizations for 2nd and 3rd trials are based on the 1st trial of Imaging.

If re-imaging is requested (different mag, different exposure, etc.,) a $100 fee is accessed for reimaging the sample with the presence of the user. A 1 hr of T12 time and staff time is dedicated for this based on staff and Instrument availability.

 Training Services (Internal and External):

 

Monday

Tuesday

Wednesday

Thursday

T12

1PM-5PM

1PM-5PM

N/A

N/A

Tf20

N/A

N/A

1PM-5PM

1PM-5PM

Vitrobot

10AM-12PM

10AM-12PM

N/A

N/A

Cryo Holders

N/A

N/A

10AM-12PM

10AM-12PM

 

T12 Training:

T12 can be used for both Negative stain EM as well as Cryo-EM.

T12 cannot be used for thin sections.

For negative staining, recommended grids are Thin carbon 300 mesh grids. Do not use Formvar coated carbon grids for negative stain TEM.

Training Fee                                                            - one-time fee per person

Instrument time                                                       - 8 hrs (split between 2 users)

login to iLab and under Request Services click to expand CryoEM screening lab and under T12 Microscope Training click Initiate request.

Training covers:

  1. Glow discharging grids, its theory and purpose.
  2. Negative staining method, its theory, purpose, QA and references.
  3. T12 SOP, alignments and theory of basic electron optics.

After the training is finished, the user will be semi-independent and will be allowed to sign up for the microscope between 9 AM to 5 PM. The user will eventually be classified as Independent user and can access the instrument without any assistance from staff. Once classified as Independent user, the user will gain access to the instrument from 8 AM to 10 PM.

If the user fails to learn the operation of the microscope, the user will be retrained on a one-to-one basis and all of the aforementioned charges will be accessed.

Tf20 Cryo-EM Training:

T12 training is a prerequisite for Cryo-EM Training.

Training is done in a group of 2 users

Tf20 should be used for Cryo-EM as well as Cryo-ET only.

Training Fee (one time only)                                   - one-time fee per person

TF20 Instrument charge                                          - 8 hrs (split between 2 users)

Vitrobot charge                                                        - 2 hrs (split between 2 users)

Pumping Station, Cryo Holders (no charge)           - 1 hr  (split between 2 users)

login to iLab and under Request Services click to expand CryoEM screening lab and under TF20 Microscope Training click Initiate request.

Training Covers:
  1. Electron optics theory, Contrast transfer, TF20 SOP, QA and references.
  2. Safety handling of liquid nitrogen, ethane.

After the training is finished, the user will be semi-independent and will be allowed to sign up for the microscope between 9 AM to 5 PM. The user will eventually be classified as Independent user and can access the instrument without any assistance from staff. Once classified as Independent user, the user will gain access to the instrument from 8 AM to 10 PM.

If the user fails to learn the operation of microscope, the user will be retrained on a one-to-one basis and all of the aforementioned charges will be accessed.