The Fluorescence Correlation Spectroscopy/Confocal Imaging Facility of the Department of Biochemistry and Molecular Biophysics is open to Washington University researchers. The facility has a Zeiss LSM 510 microscope with a ConfoCor 2 head for fluorescence correlation spectroscopy (FCS). FCS measures the dynamics of labeled biomolecules at sub-nanomolar concentrations inside living cells, on the cell membrane or in free solution. Diffusion coefficients, rates of directed transport, rates of chemical reactions or conformational changes and the number of fluorescent molecules in the observation volume (~femtoliter) can be determined. With appropriate labeling, cross-correlation can measure the rates and extents of molecular associations.
The LSM 510 is a state-of-the-art confocal laser-scanning microscope. An argon ion laser and two Helium/Neon lasers allow excitation over the blue-red visible spectrum, and a mercury lamp extends the excitation range into the near UV. Two fluorescence detectors and a transmitted light detector are supplemented by a META detector head which provides 32 additional wavelength-selective fluorescence channels. The linear unmixing capability of the Zeiss software can discriminate emission spectra separated by only a few nanometers.
The Details
Costs
$25/hr; nights (6:00PM-8:00AM) and weekends are half price.
Training costs are $50/hour and will vary according to the needs of the
researcher - the more you already know, the less you pay for training, but
there is a half-hour minimum training/set-up/check-out charge. Technical
assistance and consultation (beyond quick-and-easy fixes) are also $50/hour.
Annual licenses are also available. (Got some great science to do but that
last study section was composed of a bunch of stiffs who didn't just didn't
get it, so you're in between grants?? Contact us and we can probably work something out.)
Sign-up policy
To sign up for a time slot, go to the sign-up calendar. Contact Tony Pryse for a username/password if you don't already have one. If you cancel at least 24 hours
before the start of your time, no charges will be levied. Within 24 hours,
the time will be charged at ½ price unless it can be filled by someone else.
Administration
Steering Committee: Professors Elliot Elson and
Carl Frieden
Management: Tony Pryse (362-3345, email)
Useful Links
LSM 510 Manual (Manuals in pdf, will open in new window)
Chapters 1-4 of the LSM manual,
The only two chapters of
interest to most people are: 3. Introduction to Laser Scanning Microscopy. "Here
you will find an introduction to Laser Scanning Microscopy, with an explanation
of the principles of confocal imaging. The section also outlines the ways to
present LSM image series in three dimensions, and introduces you to the
performance features of your LSM 510." and 4. Quickstart
Chapter 5 of the LSM manual,
Long chapter, but it's all in here. 5 Operation in Expert Mode "In the Operation
section you
will find the most important steps and procedures of the LSM menu structure. The
step-by-step description how to get an image will be shown by typical
application examples."
Chapters 6-10 of the LSM manual
Most relevant chapters: 7. Routine Mode and Tools "This section
contains a description of the Routine Mode for scanning images using the LSM
scanning module and the use of the tools for setting the microscope." 8. 3D for
LSM 510 "This section contains a description of the LSM 3D software package
(basic program and addons). At the same time, all functions and settings are
presented in a systematic form and in the order in which they can be reached
from the basic menu via sub-menus and dialog boxes."
FCS/ConfoCor Manuals
Chapters 1-4,
Chapter 5,
and Chapters 6-10.
FCS papers
In the begining ...
More recent work:
Confocal microscopy sites
| Biochemistry | Medical School | Wash U |