Cryo-Em

Overview

Recent technological advances implemented in electron microscopes have allowed the visualization of biologically important molecules with unprecedented details. In order to bring those advances to applications for the benefit of humankind in the continuous fight against disease, La Jolla Institute decided to set up a state of the art electron cryomicroscopy facility for the benefit of local researchers. In this facility we can collect images of unstained specimens, either purified or in situ, and help with the necessary image processing to extract relevant information. The facility offers two state of the art transmission electron microscopes, a dual-beam electron microscope for the preparation of cellular lamellae, a fluorescence microscope to find the areas of interest in frozen cells, and ample sample preparation facilities.

Ruben Diaz Avalos, Ph.D.

Cryo-EM Core Director

Services

Our facility is staffed by highly trained personnel, which can assist in all aspects of the workflow for any project involving electron cryomicroscopy, from specimen preparation to data processing.

Commercial partners are welcome to use our services. Please call us for consultation and pricing.

Instrumentation

Titan Krios Electron Microscope
This state of the art electron microscope is located in a special, dehumidified room, with the microscope sitting on a piezoelectric anti-vibration table. It can be operated at either 200 or 300kV of accelerating voltages, and it is equipped with an X-FEG, which is a highly coherent Schottki type electron emitter. The images are collected on a Gatan K3 direct electron detector, after passing through a BioQuantum energy filter. Data can be collected using native ThermoScientific software for image acquisition (EPU and Tomo), or SerialEM.
Titan Halo Electron Microscope
This microscope can be operated at 200 or 300kV of accelerating voltage. The electrons are provided by an X-FEG, Schottki type field emission gun, and after traveling though the Titan type column, the image is detected with a direct electron Falcon3 camera, which has a superb quantum response. This microscope uses a side-entry holder, which allows for a wide variety of specimens that can be imaged.
Aquilos Dual Beam Microscope
This electron microscope is equipped with both an electron beam and a Gallium ion beam. The Gallium beam is used to mill away the excess material in order to create thin lamellae from frozen cells, for further visualization in the transmission electron microscopes. The electron beam is typically used to monitor the milling process, but the microscope can also be used as a typical scanning electron microscope.
Leica Fluorescence CLEM
This microscope allows the visualization of frozen cells, to determine, by the use of a fluorescent signal, the regions of interest where excess material has to be removed for high-resolution inmaging. The coordinates of the regions of interest can be saved and transfered to the other instruments down the pipeline, making for a considerably smoother workflow.
Leica Carbon Coater
Ultra thin carbon films are the bread and butter of electron microscopy. In order to produce the highest quality carbon films, at LJI we have the availability of an e--beam carbon evaporator, which produces the finest carbon films.
Vitrobot
In order to produce specimens suitable for imaging of molecules in solution, al the electron cryomicroscopy facility at LJI we have three different options of plunge freezing equipment. The facility has an FEI vitrobot, a Leica GP2, as well as a manual plunger. The users can decide which one to use, according to their preference. These instruments for sample preparation are housed in a dehumidified sample prep room, which is readily accessible.
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