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Light sheet and confocal microscopy

The Department of Biology possesses one confocal microscope, Leica SP8 DLS, that is also a light sheet microscope. The microscope is available – against a user's fee after initial training.

Light sheet microscopy

Light sheet microscopy – also referred to as single plane illumination microscopy (SPIM) is a is a gentle way of imaging sensitive samples or fast biological processes in vivo. The specimen is illuminated only in a single plane at a time and detected from the perpendicular direction. Since there is no out-of-focus excitation, phototoxic effects are reduced to the focal plane. Light sheet imaging has intrinsic optical sectioning. By moving the sample through the light sheet, 3D images of a specimen can be recorded. With this technique large 3D volumes are captured at a much higher speed but slightly lower resolution compared to confocal imaging.

Two persons sitting at a light sheet and confocal microscopy in a dark room
Leica SP8 DLS Photo: Erling Jirle

Leica SP8 DLS specifications

Our Leica SP8 DLS (inverted microsocpe) is equipped with:

Light source: five lasers with wavelenghts:

  • 405 nm
  • Solid state 488 nm
  • Solid state 514 nm
  • Solid state 552 nm
  • Solid state 638 nm

Detectors: two PMT:s (photomultiplier tubes), one HyD GaAsP-spectral detector, one PMT for transmitted light
Objectives for confocal imaging: 10x/0.3, 20x/0.75 IMM, 25x/0.95W, 63x/1.4 Oil, 63x/1.3 GLYC
Objectives and mirrors for light sheet microscopy: imaging 25x/0.95W DLS, 10x/0.3W DLS, illumination 2,5x/0.07, 5x/0.15, DLS TwinFlect 2.5mm, DLS TwinFlect 5mm
Software: LAS X with HyVolution, Dye Finder, 3D Visualisation, Microlab (for FRAP, FLIP and FRET) and Huygens Base Package for Deconvolution

Confocal microscopy

Confocal microscopy – or to be more precise: confocal laser scanning microscopy (CLSM) – is a method that is particularly suited for microscopy of fluorescent or reflecting specimens. The confocal microscope eliminates out-of-focus light, and creates a clear view of structures in the focal plane – also in specimens that are to thick for conventional fluorescence microscopy.

If you acquire a series of confocal images at different depths in a thick specimen, the image series can be used for 3D analysis and 3D rendering and you can visualise structures that could not be distinguished in conventional light microscopy.

Page Manager:

Contact information

Ola Gustafsson

Ola Gustafsson
Res.Sc.
Functional zoology

Telephone: +46 46-222 93 43
E-mail: Ola [dot] Gustafsson [at] biol [dot] lu [dot] se

Acknowledgements

If you are publishing an article with results obtained with the microscopes belonging to the Department of Biology, please make sure to acknowledge us:

"We acknowledge the Microscopy Facility at the Department of Biology, Lund University".

Downloads & links

Recommended websites for detailed info about confocal microscopy and other microscopy techniques:

Light Sheet Microscopy

Confocal microscopy

Other