Leica SP8 Resonant Scanning Confocal

The Leica SP8 is a high speed, high resolution microscope for investigating dynamic processes in living cells.

Overview

The Leica SP8 Resonant Scanning Confocal microscope is suitable for live cell imaging with subcellular resolution. The system contains a resonant scanning confocal mode allowing for fast imaging of dynamic cellular processes 
 
The system is compatible with most sample holders and contains all necessary accessories for environmental control, for example, CO2, humidity and temperature.

Examples of work
  • Leica SP8 Resonant Scanning Confocal microscope at the Institute
    Leica SP8 Resonant Scanning Confocal microscope
  • Microsopy image of blood vessels
    Researchers have used the Leica SP8 to study blood vessels endothelias cells in the small intestine, to better understand how blood vessels develop. Credit: Evelyn Trounson, Zoe Grant, Leigh Coultas
  • Microscopy image of stained tissue
    This image, captured on the Leica SP8, shows a section of tissue staining for different proteins that play essential roles during development. Credit: Zoe Grant
  • Microscopy image of myoblast cells
    Researchers study the genes involved in myogenesis (a process of muscle formation) to develop therapies for treating disorders such as muscular dystrophy. In this image myoblast cells are fluorescently stained with DAPI (yellow) to stain the nucleus and with Sidt2-FITC labelled antibody (pink) to stain the cytoplasm. Credit: Marilou Barrios
  • Microscopy image of breast cancer
    Fluorescent marking can be used to trace how a single rogue cell can give rise to the many cells in a tumour. While studying the development of breast cancer, researchers found these spectacular fluorescent hairs forming luminous roads. Imaged on a Leica SP8 confocal microscope and processed using Imaris software (Bitplane). Credit: Caleb Dawson
  • Microscopy image of immune cell activity
    In this image masses of immune cells (macrophages) are indicated in bright green. The lone blue ring is a cell nucleus, all that remains after a macrophage has detected and consumed a threat (bacteria or viruses). The adjacent red dot is an ASC speck, which indicates that the macrophage called for backup in its final moments. Credit: Dom De Nardo

Objectives

Magnification 10x 20x 40x 40x 63x 100x
NA 0.4 0.75 1.3 1.1 1.4 1.4
Immersion media Air Water/Glycerol Oil Water Oil Oil
XY-resolution (µm) (@ 488 nm) 0.744 0.396 0.229 0.270 1.212 0.212
Z-resolution (µm) (@ 488 nm) 6.099 2.311 0.876 1.974 0.755 0.755
Working distance (mm) 2.2 0.68 0.24 0.65 0.140 0.28

Light sources  

  • Diode laser – 405 nm 
  • Argon laser – 458 nm, 488 nm and 514 nm 
  • Diode pumped solid state laser – 561 nm 
  • HeNe Laser – 594 nm 
  • HeNe laser – 633 nm  

Detection sources  

  • 3 x PMT 
  • 2 x HyD detector

Technological specifications (capabilities) 

Available modalities 

  • Z-stack 
  • Time-series 
  • Tile scan 
  • Multi positions 
  • FRAP 

Additional hardware 

  • FRAP

Unique features 

  • Resonant scanner for fast imaging 
  • FRAP module 

Limitations 

  • Photobleaching 
  • SNR using resonant scanner is poor – requires averaging for low signal

Acknowledgement

This microscope was purchased with the generous support of the Walter and Eliza Hall Trust.