Leica Microsystems - TCS SP5 Spectral Confocal

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TCS stands for True Confocal Scanner and SP5 for the 5 SpectroPhotometer channels this system is equipped with. The microscope is a Leica DMI 6000 CS inverted microscope with a motorized XY-stage and Z-drive motor focus on the microscope stand (100 nm step size) as well as a SuperZ Galvo stage (1500 µm travel range) for higher speed and accuracy. An incubator is fitted on the microscope stage and CO2 can be piped into this chamber for live cell imaging.

In addition to standard PMTs at position 1, 3 and 5 this system has very sensitive HyD SP GaAsP-detectors at position 2 and 4 which combine high-sensitivity with low noise.

Lasers: 50 mW diode 405 nm 100 mW Argon laser 458, 476, 488, 496 and 514 nm 10 mW DPSS 561 nm 2 mW HeNe 594 nm 10 mW HeNe 633 nm

The SP5 uses an acousto-optical tunable filter (AOTF) to control the excitation laser light and an acousto-optical beam splitter (AOBS) instead of filter blocks to send the excitation laser light to the sample and to reflect the emitted light from the sample into the scan head. This is a more light efficient way than using glass di- or multichroic mirrors and filters. Besides the 5 detectors for detection of fluorescence light there is one detector for the detection of transmitted light. Keep in mind that light going to the transmitted light detector does not pass the pinhole and therefore is not one optical z-section!

The system has been upgraded with two HyD SP GaAsP-detectors with improved sensitivity in August 2012.