Cell Sciences Imaging Facility
Laser Scanning Confocal
History | System specifications | Manuals/Docs |    

 

Leica provides a free, downloadable LCS Lite image editor capable of opening and editing Leica LCS program generated experiments.

History

At the beginning of 2004, the CSIF purchased, with a NIH Shared Instrument Grant (PI Minx Fuller) a Leica SP2 AOBS Confocal Laser Scanning Microscope. The SP2 AOBS utilizes state of the art Acousto-Optical Beam Splitters (AOBS) in place of traditional glass di- or multichroics allowing user selection of reflecting excitation and transmitting emission wavelengths. The confocal is equipped with a prism spectrometer detector which allows for spectral scanning of emission profiles as well as selection of emission detection bandwidth. This allows spectral detection and un-mixing of overlapping spectrum, e.g eCFP, eGFP, and eYFP expressed in the same cell. These features make the new Leica confocal highly sensitive, flexible and, in addition to excelling in imaging fixed samples, is suited for live cell studies utilizing expressed FP-fusion proteins.

System specifications

Microscope - DM IRE2 inverted microscope.  

Lasers - HeNe 633nm/10mW, HeNe 594nm/2mW, HeNe 543/1.2 mW, Ar 458nm/5mW, 476nm/5mW, 488nm/20mW, 514nm/20mW, Blue diode 405nm

Objectives available:
Magnification N. A. Resolution x,y/z (nm) Working Distance (um)
HC PL Fluotar 10X, air 0.30 650, 4770 11000
HC PL APO 20X, H2O/Glycerol/oil 0.70 280, 1290 2500
HCX PL APO 40X, oil 1.25-0.75 160, 335 100
HCX PL APO 63X, H2O 1.2 w Corr. 140, 236 220
HCX PL APO 63X,oil 1.32-0.60 140, 236 100
PL APO 100X, oil 1.4-0.70 163, 290 90

Detectors:

  • 4 fluorescent photomultiplier tubes (PMT)
  • 1 transmitted light detector

Software: Leica Confocal, v 2.5, build 1347

  • controls all scanner and image acquisition parameters
  • provides tools for organizing and archiving image data sets
  • measuring and processing tools include colocalization and object counting
  • FRET and FRAP modules

Computers:

  • Leica-confocal, Ashlar and St Francis PCs, OS: Windows XP

These computers run Leica software. Leica-PC is interfaced with the confocal and is for acquisition of images. St. Francis and Ashlar are the processing computers for analyzing data.

Manuals and Technical Documents

To obtain a copy of the CSIF user manual for this system, you will need to have Adobe Acrobat reader installed on your computer.

Download CSIF user manual - Leica SP2 AOBS


Download Leica application note: custom_templates


Download Leica application note: customize_profile


Download Leica application note: customize_toolbars


Download Leica application note: Lambda_Scan

Download CSIF server information sheet - explains where data is stored, how long it is stored, and how to ftp from the CSIF computers to your computer.