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Further Information on the Leica SP5 Scanning Confocal
The SP5 is a scanning confocal with all of the bells and whistles. This system has 4 sensitive detectors enabling the simultaneous imaging of up to 4 different wavelengths. It has 3 lasers providing 7 excitation wavelengths and a prism spectrometer to control emission wavelengths to make this a very versatile system compatible with most combinations of fluorophores.
Features of the Leica SP5 Scanning Confocal
- Leica DMI 6000 CS inverted microscope platform)
- 7 lines of laser illumination (458,476,488,496,514,561 and 633nm) can excite from CFP to CY-5 fluorophores
- Can simultaneously image fluorescence and brightfield or DIC
- FRAP or FRET fully supported with rapid kinetic analysis
- Controlled by Leica LAS software
Sample Preparation Guidelines
Just a few things to mention to get the most out of the Leica:
- There is no UV laser to excite DAPI, so we can't image DAPI staining. We recomend DRAQ5 for staining DNA and can provide aliquots at cost for you to try.
- Always use #1.5 coverslips!
- A single coverslip in the middle of a slide works the best. This is an inverted scope. The coverslip will face down when imaging. You will need some space on either end of the slide to fit into the slide holder.
Listed here is a collection of documents, links and resources that may help you use the SP5. If you have the need or suggestions for other information please contact and let us know at firstname.lastname@example.org.
- SP5 User manual
- LAS-AF LITE software. This is a free but limited version of the imaging software that will work for viewing, exporting and some processing of your .lif files. Works on PC only.
- Application Notes
- Ruppel, N.J., C.A. Logsdon, C.W. Whippo, K. Inoue, and R.P. Hangarter. 2011. A mutation in Arabidopsis seedling plastid development1 affects plastid differentiation in embryo-derived tissues during seedling growth. Plant Physiol. 155:342-53.
- Whippo, C.W., P. Khurana, P.A. Davis, S.L. DeBlasio, D. DeSloover, C.J. Staiger, and R.P. Hangarter. 2011. THRUMIN1 is a light-regulated actin-bundling protein involved in chloroplast motility. Curr Biol. 21:59-64.
- Straiker, A., J. Wager-Miller, S.S. Hu, J.L. Blankman, B.F. Cravatt, and K. Mackie. 2011. COX-2 and FAAH can regulate the time course of depolarization induced suppression of excitation. British journal of pharmacology.
- Mowery, T.M., R.M. Sarin, K.S. Elliott, and E.G. P. 2011. Nerve injury-induced changes in GABA(A) and GABA(B) sub-unit expression in area 3b and cuneate nucleus of adult squirrel monkeys: Further evidence of developmental recapitulation. Brain research. 1415:63-75.
- Lucas, J.R., S. Courtney, M. Hassfurder, S. Dhingra, A. Bryant, and S.L. Shaw. 2011. Microtubule-Associated Proteins MAP65-1 and MAP65-2 Positively Regulate Axial Cell Growth in Etiolated Arabidopsis Hypocotyls. Plant Cell. 23:1889-903.
- Gu, Y., and R.W. Innes. 2011. The KEEP ON GOING protein of Arabidopsis recruits the ENHANCED DISEASE RESISTANCE1 protein to trans-Golgi network/early endosome vesicles. Plant Physiol. 155:1827-38.
- Christiansen, K.M., Y. Gu, N. Rodibaugh, and R.W. Innes. 2011. Negative regulation of defence signalling pathways by the EDR1 protein kinase. Molecular plant pathology.
- Chang, K.T., R.F. Niescier, and K.T. Min. 2011. Mitochondrial matrix Ca2+ as an intrinsic signal regulating mitochondrial motility in axons. Proceedings of the National Academy of Sciences of the United States of America. 108:15456-61.
- Bird, S.P., and L.A. Baker. 2011. An abiotic analogue of the nuclear pore complex hydrogel. Biomacromolecules. 12:3119-23.
- Jacob, Y., H. Stroud, C. Leblanc, S. Feng, L. Zhou, E. Caro, C. Hassel, C. Gutierrez, S.D. Michaels, and S.E. Jacobsen. 2010. Regulation of heterochromatic DNA replication by histone H3 lysine 27 methyltransferases. Nature.
- Shu-Jung Hu, S., A. Arnold, J.M. Hutchens, J. Radicke, B.F. Cravatt, J. Wager-Miller, K. Mackie, and A. Straiker. 2010. Architecture of cannabinoid signaling in mouse retina. J Comp Neurol. 518:3848-66.
- Maqbool, S.B., S. Mehrotra, A. Kolpakas, C. Durden, and B.R. Calvi. 2010. Transcriptional activation by the Myb-MuvB and E2F1 / Dp transcription factors is dampened in Drosophila endocycling cells. Journal of Cell Sci. (in press).
- Mishra, M., A. Oke, C. Lebel, E.C. McDonald, Z. Plummer, T.A. Cook, and A.C. Zelhof. 2010. Pph13 and Orthodenticle define a dual regulatory pathway for photoreceptor cell morphogenesis and function. Development.
- Hughes, H.V., E. Huitema, S. Pritchard, K.C. Keiler, Y.V. Brun, and P.H. Viollier. 2010. Protein localization and dynamics within a bacterial organelle. Proc Natl Acad Sci U S A. 107:5599-604.
- Chang, K.T., and K.T. Min. 2009. Upregulation of three Drosophila homologs of human chromosome 21 genes alters synaptic function: implications for Down syndrome. Proc Natl Acad Sci U S A. 106:17117-22.
- Straiker, A., S.S. Hu, J.Z. Long, A. Arnold, J. Wager-Miller, B.F. Cravatt, and K. Mackie. 2009. Monoacylglycerol lipase limits the duration of endocannabinoid-mediated depolarization-induced suppression of excitation in autaptic hippocampal neurons. Mol Pharmacol. 76:1220-7.
- Straiker, A., and K. Mackie. 2009. Cannabinoid signaling in inhibitory autaptic hippocampal neurons. Neuroscience. 163:190-201.