SP5 Spectral Confocal, Multiphoton, HC Screening

Leica TCS SP5 MP system

Inverted spectral confocal microscope for advanced 3D/4D/5D imaging, Time Lapse, High-Content Screening and Multiphoton imaging.

The system performs High Content Screening with the Leica Matrix HCS-A Software module.

The system can also perform Intelligent Screening  with our custom ImageJ interface which controls the SP5 acquisition through the CAM module (Leica).

Contact Lídia Bardia and Anna Lladó to use this instrument:  microscopy(at)irbbarcelona.org


 Technical Features


 Feature         Specifications Comments
 Laser Illumination
405nm diode laser
Argon laser (458-476-488-496-501-514nm)
561nm DPSS laser
594nm HeNe laser
633nm HeNe laser

IR laser MaiTai Wide Band (710nm-990nm)
 Dapi-like
 CFP; GFP; Cy2; Alexa 488-like (A488); YFP
 RFP; Cy3; A568-like
 A594; TxRed-like
 Cy5; A647-like

 Efficient on DAPI-GFP-YFP like excitation spectra
 possible, but less efficient in the red spectral range.
 Microscope DMI 6000
 Inverted microscope
 Acquisition Software
 LAS AF
 version 2.4.1 + Matrix HCS-A
 Detectors 3 PMTs + 2 HyDs + 1 PMT for transmission detection
 2 external Non Descanned detectors for multiphoton acquisition
 You can create a sequential  setting with more than 10 channels
Scanning 2D (x-y; x-t; x-z)
 3D (x-y-z; x-y-t; x-y-lambda)
 4D (x-y-z-t; x-y-t-lambda)
 
 Resolution
 from 16x16 up to 8192x8192 pixels
 
 Image depth
 8-12-16 bit
 
 Zoom
 1x-64x range although minimal zoom depends on speed:
 1.7x for Resonant Scanner (RS) mode and 700Hz, 3x for 1.000Hz and 6x for 1.400Hz)
 
 Speed In confocal:
     max 2.800Hz (bi-directional mode)
 In RS mode:
     16.000Hz (bi-directional mode)
 
 Sample mounting
 For an inverted microscope options for slides, 35mm petri dishes, see also Accessories
 Stage
Maerzhaueser Motorized stage for  X-Y multiple fields Images
  Multiposition 'Mark and Find' with LAS-AF  and
  Mosaic/MultiWell  options with Matrix Software Module
 Widefield Fluorescence
Filters
             I3:        Ex.    BP 450-490
                        BS     510
                        Em.   LP 515
            N2.1:                            Ex.    BP 515-560
                                                BS     580
                                                Em.   LP 590 
            UV:        Ex.    BP 340-380
                         BS     400
                         Em.   LP 425 

  GFP-like


  RFP-like


  DAPI-like
Objective lenses
 
    10x/0.40 CS                      HC PL APO
         
    20x/0.70 CS                      HC PL APO

    25x/0.95 W dipping, long work. dist. for IR MP imaging.
   

    40x/1.25-0.75 Oil CS         HCX PL APO

    63x/1.40-0.60 Oil Lbd BL  
HCX PL APO

    63x/1.3Glyc Corr CS 21º     HCX PL APO

    63x/1.20 W Corr/0.17CS    HCX PL APO
 
    Dipping lenses available for long working distances
    see Accessories
 Environment Temperature control over whole microscope body, CO2 control locally (closed stage insert)
 


 Applications


 Confocal Microscopy
 Fixed sample scanning
 Colocalization analysis
 Live Imaging
 Transmission detection
 Bright Field and DIC
 Resonant Scanner
 16.000Hz speed
 Multiphoton Fixed and live thick samples
    (Zebra fish, Drosophila embryos, Tissue sections)
 Second Harmonic Generation
 High Content Screening HCS-A Matrix software (Multiwell, slides, Tissue sections)
 FRAP/FLIP Flymode FRAP, derivated techniques of photobleaching (FLIP etc...)
 FRET Sensitized Emission /  Acceptor Photobleaching
 Photoactivation
 like PA-GFP protein
 Photoconversion from green to red like          Dendra2 or EOS proteins


 Special Applications (open projects in development at ADM)


 Intelligent Screening,     ImageJ controls the microscope through CAM, development by ADM in collaboration with Leica.

 Micropatterned Cells:     Screening and Analysis with Cytoo
® Chips, open solutions to the analysis of cells on CytooChips,
                                     development by ADM in collaboration with Cytoo
®