Leica SP8 Resonant Scanning Confocal

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

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.

Equipment booking

Above: Blood vessels sprouting out of a piece of lab-grown bone can be live-imaged, allowing researchers to better understand how blood vessel growth is controlled. Credit: Zoe Grant

Above: Blood vessels converging at the centre of the retina. Credit: Sabrina Lewis and Leigh Coultas

Objectives

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

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

Above: Researchers are studying the architecture and cellular composition of the structures that branch from the end of mammary ducts – the sites where breast cancer usually arises. Credit: Bianca Capaldo

Above: 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

Technological specifications (capabilities)

Available modalities

Z-stack
Time-series
Tile scan
Multi positions
FRAP

Additional hardware

FRAP

Above: 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

Above: 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

Unique features

Resonant scanner for fast imaging
FRAP module

Above: 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

Above: 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

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.

Above: Blood vessels at the edge of the retina. Credit: Sabrina Lewis and Leigh Coultas

Above: Migrating blood vessels (red) and blood cells (white) are observed in the study of how platelets affect brain bleeds/stroke. Credit: Alison Farley