• Source: Imaging cycler microscopy

An imaging cycler microscope (ICM) is a fully automated (epi) fluorescence microscope which overcomes the spectral resolution limit resulting in parameter- and dimension-unlimited fluorescence imaging. The principle and robotic device was described by Walter Schubert in 1997 and has been further developed with his co-workers within the human toponome project. The ICM runs robotically controlled repetitive incubation-imaging-bleaching cycles with dye-conjugated probe libraries recognizing target structures in situ (biomolecules in fixed cells or tissue sections). This results in the transmission of a randomly large number of distinct biological informations by re-using the same fluorescence channel after bleaching for the transmission of another biological information using the same dye which is conjugated to another specific probe, a.s.o. Thereby noise-reduced quasi-multichannel fluorescence images with reproducible physical, geometrical, and biophysical stabilities are generated. The resulting power of combinatorial molecular discrimination (PCMD) per data point is given by 65,536k, where 65,536 is the number of grey value levels (output of a 16-bit CCD camera), and k is the number of co-mapped biomolecules and/or subdomains per biomolecule(s). High PCMD has been shown for k = 100, and in principle can be expanded for much higher numbers of k. In contrast to traditional multichannel–few-parameter fluorescence microscopy (panel a in the figure) high PCMDs in an ICM lead to high functional and spatial resolution (panel b in the figure). Systematic ICM analysis of biological systems reveals the supramolecular segregation law that describes the principle of order of large, hierarchically organized biomolecular networks in situ (toponome). The ICM is the core technology for the systematic mapping of the complete protein network code in tissues (human toponome project). The original ICM method includes any modification of the bleaching step. Corresponding modifications have been reported for antibody retrieval and chemical dye-quenching debated recently. The Toponome Imaging Systems (TIS) and multi-epitope-ligand cartographs (MELC) represent different stages of the ICM technological development. Imaging cycler microscopy received the American ISAC best paper award in 2008 for the three symbol code of organized proteomes.




Citations






References


"3D all-organelle real time visualization of a single cell"
"Visualizing the protein-DNA network code inside the cell nucleus"




Further reading


Abott, A (12 October 2006). "Research highlights". Nature. 443 (7112): 608–609. Bibcode:2006Natur.443..608.. doi:10.1038/443608a.
Ademmer; Ebert; Müller-Ostermeyer; Friess; Büchler; Schubert; Malfertheiner (April 1998). "Effector T lymphocyte subsets in human pancreatic cancer: detection of CD8+ CD18+ cells and CD8+ CD103+ cells by multi-epitope imaging". Clinical and Experimental Immunology. 112 (1): 21–26. doi:10.1046/j.1365-2249.1998.00546.x. PMC 1904939. PMID 9566785.
Barysenka, Andrei; Dress, Andreas W.M.; Schubert, Walter (1 September 2010). "An information theoretic thresholding method for detecting protein colocalizations in stacks of fluorescence images". Journal of Biotechnology. 149 (3): 127–131. doi:10.1016/j.jbiotec.2010.01.009. PMID 20100525.
Bedner, Elzbieta; Du, Litong; Traganos, Frank; Darzynkiewicz, Zbigniew (1 January 2001). "Caffeine dissociates complexes between DNA and intercalating dyes: Application for bleaching fluorochrome-stained cells for their subsequent restaining and analysis by laser scanning cytometry". Cytometry. 43 (1): 38–45. doi:10.1002/1097-0320(20010101)43:1<38::AID-CYTO1017>3.0.CO;2-S. PMID 11122483.
Berndt, Uta; Philipsen, Lars; Bartsch, Sebastian; Hu, Yuqin; Röcken, Christoph; Bertram, Wiedenmann; Hämmerle, Marcus; Rösch, Thomas; Sturm, Andreas (2010). "Comparative Multi-Epitope-Ligand-Cartography reveals essential immunological alterations in Barrett's metaplasia and esophageal adenocarcinoma". Molecular Cancer. 9 (1): 177. doi:10.1186/1476-4598-9-177. PMC 2909181. PMID 20604962.
Bhattacharya, Sayantan; Mathew, George; Ruban, Ernie; Epstein, David B. A.; Krusche, Andreas; Hillert, Reyk; Schubert, Walter; Khan, Michael (3 December 2010). "Toponome Imaging System: Protein Network Mapping in Normal and Cancerous Colon from the Same Patient Reveals More than Five-Thousand Cancer Specific Protein Clusters and Their Subcellular Annotation by Using a Three Symbol Code". Journal of Proteome Research. 9 (12): 6112–6125. doi:10.1021/pr100157p. PMID 20822185.
Bode, Marcus; Irmler, Martin; Friedenberger, Manuela; May, Caroline; Jung, Klaus; Stephan, Christian; Meyer, Helmut E.; Lach, Christiane; Hillert, Reyk; Krusche, Andreas; Beckers, Johannes; Marcus, Katrin; Schubert, Walter (March 2008). "Interlocking transcriptomics, proteomics and toponomics technologies for brain tissue analysis in murine hippocampus". Proteomics. 8 (6): 1170–1178. doi:10.1002/pmic.200700742. PMID 18283665. S2CID 20000272.
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Bonnekoh, Bernd; Malykh, Yanina; Böckelmann, Raik; Bartsch, Sebastian; Pommer, Ansgar J.; Gollnick, Harald. (2006). "Profiling lymphocyte subpopulations in peripheral blood under efalizumab treatment of psoriasis by multi epitope ligand cartography (MELC) robot microscopy". Eur J Dermatol. 16 (6): 623–635. PMID 17229602.
Bonnekoh, B.; Pommer, A.J.; Böckelmann, R.; Hofmeister, H.; Philipsen, L.; Gollnick, H. (2007). "Topo-Proteomic in situ Analysis of Psoriatic Plaque under Efalizumab Treatment". Skin Pharmacology and Physiology. 20 (5): 237–252. doi:10.1159/000104422. PMID 17587888. S2CID 43539332.
Bonnekoh, Bernd; Pommer, Ansgar J.; Böckelmann, Raik; Philipsen, Lars; Hofmeister, Henning; Gollnick, Harald (June 2008). "In-situ-topoproteome analysis of cutaneous lymphomas: Perspectives of assistance for dermatohistologic diagnostics by Multi Epitope Ligand Cartography (MELC)". Journal der Deutschen Dermatologischen Gesellschaft. 6 (12): 1038–51. doi:10.1111/j.1610-0387.2007.06754.x. PMID 18540979.
Coste, O.; Brenneis, C.; Linke, B.; Pierre, S.; Maeurer, C.; Becker, W.; Schmidt, H.; Gao, W.; Geisslinger, G.; Scholich, K. (10 September 2008). "Sphingosine 1-Phosphate Modulates Spinal Nociceptive Processing". Journal of Biological Chemistry. 283 (47): 32442–32451. doi:10.1074/jbc.M806410200. PMID 18805787.
Dress, Andreas W. M.; Lokot, T.; Pustyl’nikov, L. D.; Schubert, W. (January 2005). "Poisson Numbers and Poisson Distributions in Subset Surprisology". Annals of Combinatorics. 8 (4): 473–485. doi:10.1007/s00026-004-0234-2. S2CID 122237186.
Dress, Andreas; Lokot, Tatjana; Schubert, Walter; Serocka, Peter (3 October 2008). "Two Theorems about Similarity Maps". Annals of Combinatorics. 12 (3): 279–290. doi:10.1007/s00026-008-0351-4.
Ebert, Matthias P.A.; Ademmer, Karin; Muller-Ostermeyer, Frauke; Friess, Helmut; Buchler, Markus W.; Schubert, Walter; Malfertheiner, Peter (November 1998). "CD8+CD103+ T cells analogous to intestinal intraepithelial lymphocytes infiltrate the pancreas in chronic pancreatitis". The American Journal of Gastroenterology. 93 (11): 2141–2147. doi:10.1111/j.1572-0241.1998.00610.x. PMID 9820387. S2CID 19518135.
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Eckhardt, J.; Ostalecki, C.; Kuczera, K.; Schuler, G.; Pommer, A. J.; Lechmann, M. (16 November 2012). "Murine Whole-Organ Immune Cell Populations Revealed by Multi-epitope-Ligand Cartography". Journal of Histochemistry & Cytochemistry. 61 (2): 125–133. doi:10.1369/0022155412470140. PMC 3636694. PMID 23160665.
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Haars, Regina; Schneider, Abidat; Bode, Marcus; Schubert, W. (2000). "Secretion and differential localization of the proteolytic cleavage products Abeta40 and Abeta42 of the Alzheimer amyloid precursor protein in human fetal myogenic cells". European Journal of Cell Biology. 79 (6): 400–406. doi:10.1078/0171-9335-00064. PMID 10928455.
Herold, Julia; Schubert, Walter; Nattkemper, Tim W. (15 September 2010). "Automated detection and quantification of fluorescently labeled synapses in murine brain tissue sections for high throughput applications". Journal of Biotechnology. 149 (4): 299–309. doi:10.1016/j.jbiotec.2010.03.004. PMID 20230863.
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Schubert, Walter; Gieseler, Anne; Krusche, Andreas; Serocka, Peter; Hillert, Reyk (June 2012). "Next-generation biomarkers based on 100-parameter functional super-resolution microscopy TIS". New Biotechnology. 29 (5): 599–610. doi:10.1016/j.nbt.2011.12.004. PMID 22209707.
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Schubert, Walter (January 2014). "Systematic, spatial imaging of large multimolecular assemblies and the emerging principles of supramolecular order in biological systems". Journal of Molecular Recognition. 27 (1): 3–18. doi:10.1002/jmr.2326. PMC 4283051. PMID 24375580.
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Kata Kunci Pencarian:

• Imaging cycler microscopy
• Microscopy
• Photoactivated localization microscopy
• Digital holographic microscopy
• Toponomics
• Atomic force microscopy
• Toponome
• Magnetic resonance imaging
• Non-contact atomic force microscopy
• Fluorescence imaging