Confocal
Zeiss LSM510 Confocal
The LSHTM confocal microscope is an Zeiss axiovert contained within a incubation chamber, which can be temperature and CO2 regulated. The microscope has an electronic stage, and uses Zeiss LSM510 software for image acquisition. The microscope is equipped with x10, x20, x40, x63 and x100 lenses, and the following lasers;
Laser Diode: 405nm i.e. DAPI
Argon laser: 458, 488, 514nm i.e.FITC/Alexa 488
HeNe1 laser: 543nm i.e. rhodamine/CY3/Alexa 546
HeNe2 laser: 633nm i.e. CY5/Alexa 633
Laser scanning microscopy (confocal) enables images on a single plane to be taken within a larger section. It uses light from a laser to excite the sample which is passed though a pinhole which is much smaller than those used in a conventional microscope allowing light from a narrow plane of focus to pass through to the photomultiplier tube. The photomultiplier tube is an extremely sensitive device for converting photons into an electrical signal. The confocal microscope can also take multiple sections through cells to produce 3D images of both live and fixed samples.
This technique is useful for determining the subcellular location of stained molecules within the cell and to establish whether molecules collocate. Examples of applications used at the LSHTM are: observing cell component rearrangement and protein expression following exposure to an organism, cell-cell interactions, observing expression of recombinant proteins often linked to GFP within cells or parasites.
The microscope is also equipped with a Femtojet microinjection unit allowing the introduction of DNA into individual cells in culture.
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If you required further information or training please contact either; Rachel Gregory or Liz McCarthy.
Web Links:
- Molecular Probes web site-this is very informative for stains and antibodies-check out their gallery section for some fantastic images.
- Zeiss site-to download a image browser so you can access your LSM files on your own computer.
- Royal Microscopical Society
Other confocal sites
The Henry Wellcome imaging centre at Imperial College
University of Sussex center for Advanced microscopy
MRC cell imaging facility at the University of Bristol
