If you belong to an NYSBC-affiliated institution
If you are a member of any of these affiliate institutions, you
have access to both the instrumentation and staff
resources of the cryo-EM facility at the NYSBC. If you are
interested in embarking on a cryo-EM project at the NYSBC follow
these steps:
1. Read the Interested in EM?
page to determine if EM is suitable for your project
2. Contact your institution's
cryo-EM representative at NYSBC
3. Register for access to the
NYSBC Intranet
4. On the NYSBC Intranet fill out a
cryo-EM project proposal
5. A staff member of the cryo-EM facility will contact you to
schedule a meeting
back to top
If you do not belong to an NYSBC-affiliated institution
If you are not a member of any of the affiliate institutions listed
above, but would like to enquire about access to the facilities of the
NYSBC, please contact Dr. David Cowburn.
back to top
NYSBC Cryo-EM facility and resources
Instrumentation
To enable the application of the above-mentioned techniques to a variety
of biologically relevant samples, the NYSBC has four electron microscopes for
use by affiliate members: A 120kV screening microscope (JEOL 1230);
two 200kV cryomicroscopes (Tecnai F20 and JEOL 2100); and a 300kV
energy-filtered cryomicroscope (JEOL 3200). The 200kV and 300kV
instruments have field emission guns and two digital cameras, a fast
scan camera for searching and focusing, and a slow-scan camera for
image acquisition. All microscopes have computer interfaces that
communicate with specialized programs for automated acquisition of
data, which is convenient for single particle analysis and
crystallography and absolutely essential for tomography. In addition
to the microscopes, we have all necessary equipment for sample
preparation, such as two plungers to freeze crystals or macromolecular
suspensions, two high pressure freezers for tissue, a freeze
substitution machine, two ultramicrotomes, a carbon evaporator for
making sample support films and a wide variety of negative stains.
Thus, affiliates simply need to provide suitable samples and all
EM-specific sample preparation can be handled at NYSBC.
For further details see equipment .
back to top
Technical assistance
In addition to instrumentation, affiliates can take advantage of NYSBC
staff expertise in each of these four technologies. Initially, staff
will discuss the feasibility of projects and lay out a plan for the
different stages of a proposed research project. Affiliates can work
closely with NYSBC staff to evaluate the suitability of existing
samples for achieving specified goals and in implementing any unusual
experimental approaches. Staff members are also available for pilot
studies to produce preliminary reconstructions suitable for inclusion
in grant proposals. A sustained commitment from the affiliate lab is
generally required to achieve the ultimate goal and NYSBC staff
routinely provides training to individual researchers to be able to
function in the supported environment at NYSBC. At the beginning of a
project, this involves plunging samples into liquid ethane,
transferring these samples into the microscope column and collecting
the necessary images. Microscopes are aligned each morning and staff
is available for assisting in sample loading and microscope operation.
Once affiliates obtain a basic understanding of operation, a training
course is used to establish independence, thus making microscopes
available 24/7. NYSBC offers a course each fall semester in image
processing in the fall semester in which leading researchers from the
NY area together with NYSBC staff teach the principles and practice of
3D structure determination by electron microscopy. All major software
packages are installed at NYSBC and continuing guidance in their use
is available through personal interactions with our
staff.
back to top
