Leica TCS SP5
 

Confocal Microscope

 

  

Confocal microscopy offers several advantages over conventional optical microscopy, including controllable depth of field, the elimination of image degrading out-of-focus information, and the ability to collect serial optical sections from thick specimens. The key to the confocal approach is the use of pinhole to eliminate out-of-focus light or flare in specimens that are thicker than the plane of focus. This approach can improve the resolution and contrast of images.

 
               

Fig Principle of the confocal microscopy

NOBC has equipped 2 confocal microscope workstation:oldsystemLeica TCS SP5and new systemLeica TCS SP5 II. These systems are able to provide multi-color fluorescence imaging, 3D reconstruction, dynamic 3D imaging, morphology, multi-color GFP imaging, Z-section and measurement, and transmitted/incident light image acquisitions. The new systemLeica TCS SP5 IIhas equipped live cell incubation system and a DMI 6000 inverted microscope with a fully motorized stage. This high precision stage allows the user to perform tile scans, define sample positions in a mark & find mode for recall during live or time-lapse experiments.

 

Leica TCS SP5

This True Confocal Scanner (TCS) SP5 module is equipped with dichroic beam splitters on three tunable spectral detection PMTs plus a channel of transmitted light (bright field or DIC) for simultaneous or sequential fluorescence image acquisition. The SP5 has user friendly, intuitive software for ROI (Region Of Interest), sequential scanning, z-stack, and 3D visualization features such as orthogonal sectioning, 3D projection and 3D rotation. There are several tools for image improvement and quantification analysis.

  • Standard scanner includes beam park for FRAP, bleaching and photoactivaiton
  • z-Galvo stage for precision Z-sectioning
  • Inverted platform for imaging on slides or live cell dishes
  • Optical zoom for sampling to 40X
  • AOBS (acousto-optical beam splitter) plus sequential scanning capability allows for rapid sequential scanning of fluorophores with minimal bleed-through or cross-talk
  • 3D rendering and wizards for FRAP and FRET
  • AOTF (acousto-optical tunable filters) for spectral scanning, allowing separation of fluorophores with similar ranges (e.g. GFP and FITC) through spectral unmixing
  • LAS_AF Leica confocal software on Windows
  • Off-line version of LAS_AF software available on facility workstation

Lens:

Available Objectives: (All coverslips should be approximately 0.17mm - #1.5)

a.    HC PL APO 10x/0.40 PH1, WD=2.2mm, UV/405nm laser corr

  b.    HC PL APO20x/0.70 PH2, WD=0.59mm, UV/405nm laser corr

   c.   HCX PL APO 40x/1.25 PH3 Oil, WD=0.1mm, UV/405nm laser corr

d.      HCX PL APO 63x/1.40-0.60 Oil,CS lb blue, WD=0.1mm, UV/405nm laser corr

 

Lasers:
Programmable 8-channel AOBS beam splitter (AOBS=Acousto-Optical Beam Splitter)
Diod Visible Laser Merge Fiber Coupling System

  a.    laser system I: 100mW Ar-Blue Laser, 458nm, 476nm, 488nm, 496nm, 514nm

  b.       laser system II: 10mW yellow DPSS laser, 561nm

  c.       laser system III: 10mW HeNe Red Laser , 633 nm

 

405nm Laser Merge Fiber Coupling System

 a.   laser 405 nm 50 mW continuous wave, non-pulsed with AOTF

 

Leica TCS SP5 II

This True Confocal Scanner (TCS) SP5 II module is equipped with dichroic beam splitters on three tunable spectral detection PMTs plus a channel of transmitted light (bright field or DIC) for simultaneous or sequential fluorescence image acquisition. The SP5 II has user friendly, intuitive software for ROI (Region Of Interest), sequential scanning, z-stack, and 3D visualization features such as orthogonal sectioning, 3D projection and 3D rotation. There are several tools for image improvement and quantification analysis.

  • 3 chilled PMT fluorescence detectors (digital spectral definition in 1 nm increments) plus one transmitted light detector with DIC polarizer/analyzer plus prisms for most objectives available. 8- or 12-bit output, 16-bit possible in galvo mode.
  • Standard scanner includes beam park for FRAP, bleaching and photoactivaiton
  • z-Galvo stage for precision Z-sectioning
  • Inverted platform for imaging on slides or live cell dishes
  • Optical zoom for sampling to 40X
  • AOBS (acousto-optical beam splitter) plus sequential scanning capability allows for rapid sequential scanning of fluorophores with minimal bleed-through or cross-talk
  • 3D rendering and wizards for FRAP and FRET
  • AOTF (acousto-optical tunable filters) for spectral scanning, allowing separation of fluorophores with similar ranges (e.g. GFP and FITC) through spectral unmixing
  • LAS_AF Leica confocal software on Windows
  • Off-line version of LAS_AF software available on facility workstation

Lens:

Available Objectives: (All coverslips should be approximately 0.17mm - #1.5)

 1.   HC PL APO 10x/0.40 CS, WD=2.2mm, UV/405nm laser corr

  2.   HC PL APO 20x/0.70, WD=0.59mm, UV/405nm laser corr

  3.   HCX PL-APO 40x/1.25-0.75 Oil CS, WD=0.1mm, UV/405nm laser corr

4.      HCX PL APO 63x/1.40-0.60 Oil,CS, WD=0.1mm, UV/405nm laser corr

 

Lasers:
Programmable 8-channel AOBS beam splitter (AOBS=Acousto-Optical Beam Splitter)
Diod Visible Laser Merge Fiber Coupling System

  a.    laser system I: 100mW Ar-Blue Laser, 458nm, 476nm, 488nm, 496nm, 514nm

  b.              laser system II: 1mW HeNe Green laser, 543nm.

   c.    laser system III: 2mW HeNe Orange laser, 594nm.

    d.    laser system III: 10mW HeNe Red Laser , 633 nm

 

405nm Laser Merge Fiber Coupling System

 a.   laser 405 nm 50 mW continuous wave, non-pulsed with AOTF

 

Live cell incubation system

 

 

 

 

 

The difference between TCS SP5 and TCS SP5II

 

SP5 II

SP5

living cell incubator

yes

no

Scanning stage

Yes

no

Laser system

405nm Diode laser

Argon Blue-458, 476, 488, 496, 514nm

HeNe 543nm

HeNe Orange 594nm

HeNe Red 633nm

405nm Diode laser

Argon Blue-458, 476, 488, 496, 514nm

DSPP 561nm

HeNe Red 633nm

                           
                               
 
 

 

公告日:2011/07/14





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