Confocal laser scanning microscopes

SP5-Sted  Leica Sp8  Leica sp5
Leica Sp5-Sted  Leica Sp8  Leica Sp5
  

Leica TCS SP8 X WLL

  

Leica SP5-Sted  Confocal Microscope

SP5-Sted

The new Leica TCS STED CW confocal microscope is the most straightforward way to nanoscopy for research. The system reveals sub-cellular details below 80 nm – based on a purely optical approach. Sub-diffraction multicolor images provide new insights into cellular architecture. The depletion laser and the excitation lines in the visible range enable researchers to use conventional dyes such as Alexa 488, FITC and Oregon Green and established fluorescent proteins such as YFP. Dynamics of sub-cellular processes can be imaged due to this flexibility and the system's ability of fast data acquisition. This means: live cell imaging below the diffraction limit!

The following objectives are available for imaging:

  • HC PL FLUOTAR 10x 0.30
  • HC PL APO 20x 0.70 Imm Corr cs
  • HCX PL APO CS 40x 1.25-0.75 Oil
  • HCX PL APO CS 63x 1.40-0.60 Oil
  • HCX PL APO 100x 1.40 Oil (STED Objective)

A  heated chamber with CO2 controller and objective heater are available for prolonged (time lapse) imaging of live cells. 

The system is also equipped with two GaAsP hybrid detection systems (Leica HyD) for confocal imaging, which significantly improves image quality and detection efficiency.

You can find some more documentation here.

Example of STED versus Confocal image

  Confocal image

Confocal image of endoplasmic reticulum (gfp)

STED image

STED image of endoplasmic reticulum (gfp)


Leica TCS SP8

  Leica Sp8

The Leica TCS SP8 upright confocal microscope is equipped with two sensitive HyD detectors, one Photo-multiplier Tube (PMT) detector for fluorescence and an additional PMT for bright-field imaging. Images can be acquired with scan resolutions of up to 8192 x 8192 pixels. Spectral detectors permit independent real-time adjustment of the detection bandwidths in the range 380 to 800 nm for each of the confocal fluorescence detector channels, eliminating the need for emission filters. Images can be acquired in simultaneous or sequential scan modes. The four solid-state lasers provide the following laser lines: 405 nm, 488 nm, 552 nm and 638 nm. The intensity of each laser line is independently adjustable from 0-100% power using the acousto-optical-tuneable filter (AOTF). A Z-wide stage permits rapid and precise acquisition of Z-stacks. The microscope is equipped with a 10x/0.3 NA dry objective, a 20x/0.5 NA dry objective, and 40x/1.30 NA and 63x/1.40 NA Plan Apo oil immersion objectives for high resolution imaging. On this confocal microscope only microscope objective slides can be used for imaging.  

 

User Manual

 

 


Leica TCS SP5

Leica sp5

Overview

This system combines conventional and high-speed technology in one single system with the new Tandem Scanner.  For demanding multiparameter fluorescence imaging, the Leica TCS SP5 is equipped with high efficiency SP-detection (five channels simultaneously) and an AOBS for dynamic beam splitting. The system has software to measure molecular dynamics on the basis of FRAP and FLIP. Other facilities: climate controlled box, and multiphoton laser. 

 Optical Equipment
  • Hc PL APO CS 10x 0.4 dry
  • 20x dry
  • 63x Oil
  • HCX PL APO CS 40x 1.25 Oil
  • HCX PL APO Lambda blue 63x 1.4 Oil
  • HCX PL APO CS 100x 1.4 Oil
Available Laser lines
  • 405 nm
  • 458 nm
  • 476 nm
  • 488 nm
  • 496 nm
  • 514 nm
  • 561 nm
  • 594 nm
  • 633 nm
  • MP
Key Features 
  • True confocal point-scanning - real optical sectioning
  • Large field of view (21 mm)
  • 64 Megapixel images (8k x 8k)
  • Beam park data acquisition
  • Spot-bleaching FRAP
  • Zoom and pan function
  • Slow scan mode
  • Up to 5 confocal channels simultaneously
  • True confocal point-scanning - real optical sectioning
  • Large field of view (15 mm)
  • Zoom and pan function
  • 16,000 lines per second
  • 250 frames per second
  • 25 frames per second 512 x 512
  • Up to five confocal channels simultaneously 
  • Conventional and resonant Scanner in one system
  • Switch by computer controlled super precision mechanical actuator
  • Five confocal channels
  • True confocal point-scanning - real optical sectioning
  • High dynamic sensors
  • Low dark current PMTs
  • High efficiency prism spectrometer
  • Loss-free band selection
  • Free tunable emission bands
  • Five bands simultaneously
  • Dye separation: sufficient for five channels simultaneously 

Leica TCS SP8 X WLL

The newly installed Leica TCS SP8 X WLL is the only freely tuneable Confocal System.

The white light laser (WLL) source of the Leica TCS SP8 X WLL perfectly matches the spectral properties of any fluorophore excitable in the visible spectral range. Continuously tuneable excitation and detection ensure best image quality and sample protection. Up to eight excitation lines can be used – simultaneously.

  • Tuning range of 470 to 670 nm in 1 nm intervals
  • Minimum cross-talk in multiple stained specimen
  • Change of Illumination regimes within microseconds
  • Lambda Square Mapping: full spectral information by excitation/emission correlation
  • LightGate: background quenching using time-gated detection

FREELY TUNABLE CONFOCAL IMAGING WITH THE LEICA TCS SP8 X WLL.

The White Light Laser (WLL) is the optimal excitation source to image any kind of dye combination. The acousto-optical beam splitter (AOBS) is the key to selecting any wavelength from the white spectrum. Completed by the tuneable spectral detector (SP), the Leica TCS SP8 X WLL represents the only filter-free versatile confocal microscope. A two-dimensional excitation-emission spectrum characterizes the spectral properties of any fluorescent specimen. Fluorophores with either identical emission spectra or excitation spectra are immediately discriminated. This is essential for optimizing multi-colour fluorescence experiments. The highest image contrast can be obtained by LightGate. This is an innovative, filter-free method to quench unwanted signals. An adjustable time gate switches off the data collection during the white light laser pulse excluding any reflected photons from the signal. Additionally, non-wanted fluorescence can be removed by adjusting the time gate.

The new Leica TCS SP8 X WLL is furthermore equipped with a UV laser (405 nm) and it is suited for live cell imaging. Temperature and CO2 can be controlled in the software.

The software furthermore includes possibilities for 3D visualization and analysis of your images.