3D electron microscopy data can be acquired by Focused Ion Beam Scanning Electron Microscopy FIB-SEM where fine sequence of 4-8 nm increments are ablated off of a sample surface and each such surface is imaged with the SEM. At the finest resolution and with month-long stable operation, comprehensive whole cells can be acquired that transcends the limited cut section views of traditional TEM used in biology.  Several examples of such data are presented along with the potential that segmentation offers to explore and formulate biological questions. Correlative microscopy can be achieved by a cryogenic protocol where samples are vitrified, imaged with PALM or SIM at low temperatures followed by EM staining and FIBSEM.  A 3D registration procedure can keep most position errors between PALM and EM data at ~ 30 nm.  Examples validating the approach with mitochondrial and endoplasmic reticulum labels are presented along with examples showcasing how unknown vesicle types and other structures can be identified by an associated protein.

Those who wish to meet the speaker should contact Shikhar Dhingra (Fitzpatrick Lab).


The Columbia Neuroscience Seminar series is a collaborative effort of Columbia's Zuckerman Institute, the Department of Neuroscience, the Doctoral Program in Neurobiology and Behavior and the Columbia Translational Neuroscience Initiative, and with support from the Kavli Institute for Brain Science.