The Leica confocal is one of a few laser scanning confocal microscopes in the AIF.  The  Leica may be programmed to scan many different wavelength ranges.   The Leica AOBS uses variable spectral detection instead of traditional emission filters.  There are a few laser lines for excitation and the dyes are limited by these.  Any dye that is excited at one of these following wavelengths can be used:   405; 458; 476; 488; 514; 561; and 633 nm.   According to the sales literature, the lasers are:  diode 20 mW 405 nm; Ar 100 mW 457 nm, 488 nm, 514 nm; diode 10mW 561 nm; HeNe 10 mW 633 nm.

Objectives:
63X N.A. 1.4-0.60 Oil l_BL HCX PL APO
40X N.A.  1.25-0.75 Oil CS HCX PL APO
20X N.A.  0.70 1mm corr l_BL HC PL APO

Automated stage for tiling XY, XYZ or XYT  volumes with 166.6 mm of travel in Z.

Here's a picture of the new system as installed at the AIF on November 6, 2002.

Contents:

• Main manual for turn on / shut down
• Environmental control for live cell work
• A few example images

Leica Application Notes:

• FRAP
• More FRAP
• appnotes/app_note_custom_templates2.PDF
• appnotes/app_note_customize_profile2.pdf
• appnotes/app_note_customize_toolbars2.pdf
• appnotes/Sequential-Scanning-Letter.pdf
• appnotes/app_note_quantify.pdf
• appnotes/app_note_Scale_Bar_1537.pdf
• appnotes/app_note_LCS_backup.pdf

Sample method section for a paper:

Images were collected with a Leica TCS SP2 AOBS confocal microscope (Mannheim, Germany) with 25X and 60X oil immersion optics. Laser lines at 488nm and 543nm for excitation of Cy2 and Cy3 were provided by an Ar laser and a HeNe laser. Detection ranges were set to eliminate crosstalk between fluorophores.

Selected Bibliography

1. Azios, NG, Krishnamoorthy, L, Harris, M., Cubano, LA, Cammer, M, Dharmawardhane, SF.   Estrogen and Resveratrol Regulate Rac and Cdc42 Signaling to the Actin Cytoskeleton of Metastatic Breast Cancer Cells. Neoplasia. 2007 Feb; 9(2):147-158.
2. Bhatia S, Edidin M, Almo SC, Nathenson SG. (2005) Different cell surface oligomeric states of B7-1 and B7-2: Implications for signaling. Proc Natl Acad Sci U S A. 102(43):15569-15574. PMID: 16221763
3. Eugenin EA, Berman JW.  Gap junctions mediate human immunodeficiency virus-bystander killing in astrocytes. J Neurosci. 2007 Nov 21;27(47):12844-50. PMID: 18032656
4. Herskovits AZ, Davies P. (2006) The regulation of tau phosphorylation by PCTAIRE 3: Implications for the pathogenesis of Alzheimer's disease. Neurobiol Dis. 2006 Jun 9; [Epub ahead of print] PMID: 16766195
5. Lazar-Molnar E, Almo SC, Nathenson SG.  The interchain disulfide linkage is not a prerequisite but enhances CD28 costimulatory function. Cell Immunol. 2007 Apr 27; [Epub ahead of print] PMID: 17467674
6. Maxson, ME, Cook, E, Casadevall, A, Zaragoza, O.  The volume and hydration of the Cryptococcus neoformans polysaccharide capsule. Fungal Genetics and Biology. March 2007; 44(3)180-186
7. Shav-Tal Y, Darzacq X, Shenoy SM, Fusco D, Janicki SM, Spector DL, Singer RH. (2004) Dynamics of Single mRNPs in Nuclei of Living Cells. Science 304(5678):1797-1800.
8. Zhang X, Schwartz JC, Guo X, Bhatia S, Cao E, Lorenz M, Cammer M, Chen L, Zhang ZY, Edidin MA, Nathenson SG, Almo SC. (2004) Structural and functional analysis of the costimulatory receptor programmed death-1. Immunity. 20(3):337-47.