laser scanning cytometry

Summary

Summary: A scanning microscope-based, cytofluorimetry technique for making fluorescence measurements and topographic analysis on individual cells. Lasers are used to excite fluorochromes in labeled cellular specimens. Fluorescence is detected in multiple discrete wavelengths and the locational data is processed to quantitatively assess APOPTOSIS; PLOIDIES; cell proliferation; GENE EXPRESSION; PROTEIN TRANSPORT; and other cellular processes.

Top Publications

  1. Mosch B, Mittag A, Lenz D, Arendt T, Tarnok A. Laser scanning cytometry in human brain slices. Cytometry A. 2006;69:135-8 pubmed
    The Laser Scanning Cytometry (LSC) offers quantitative fluorescence analysis of cell suspensions and tissue sections. We adapted this technique to immunohistochemical labelled human brain slices...
  2. Zhao H, Albino A, Jorgensen E, Traganos F, Darzynkiewicz Z. DNA damage response induced by tobacco smoke in normal human bronchial epithelial and A549 pulmonary adenocarcinoma cells assessed by laser scanning cytometry. Cytometry A. 2009;75:840-7 pubmed publisher
    ..Multiparameter analysis by laser scanning cytometry made it possible to relate these DDR events, detected immunocytochemically, with cell cycle phase...
  3. Fueldner C, Mittag A, Knauer J, Biskop M, Hepp P, Scholz R, et al. Identification and evaluation of novel synovial tissue biomarkers in rheumatoid arthritis by laser scanning cytometry. Arthritis Res Ther. 2012;14:R8 pubmed publisher
    ..Therefore, we tested several monoclonal antibodies (mAbs) directed against cellular-surface molecules on cells likely to be involved in the pathogenesis of RA...
  4. Harnett M. Laser scanning cytometry: understanding the immune system in situ. Nat Rev Immunol. 2007;7:897-904 pubmed
    ..b>Laser scanning cytometry is an emerging technology that allows imaging and quantitative analysis of individual cells in tissues in ..
  5. Peterson R, Krull D, Butler L. Applications of laser scanning cytometry in immunohistochemistry and routine histopathology. Toxicol Pathol. 2008;36:117-32 pubmed publisher
    b>Laser scanning cytometry (LSC) is a powerful tool for qualitative and quantitative analysis of tissue sections in preclinical drug development. LSC combines the strengths of flow cytometry with tissue architecture retention...
  6. Darzynkiewicz Z, Huang X, Okafuji M. Detection of DNA strand breaks by flow and laser scanning cytometry in studies of apoptosis and cell proliferation (DNA replication). Methods Mol Biol. 2006;314:81-93 pubmed
    ..chapter describes methods to label in situ DNA strand breaks with fluorochromes for detection by flow or laser scanning cytometry. By staining DNA with a fluorochrome of another color, cellular DNA content is measured concurrently and ..
  7. Iglesias I, Bentsi Barnes K, Umeadi C, Brown L, Kandeel F, al Abdullah I. Comprehensive analysis of human pancreatic islets using flow and laser scanning cytometry. Transplant Proc. 2008;40:351-4 pubmed publisher
    Assessing islet cellular composition and beta cell viability using Flow Cytometry (FC) and Laser Scanning Cytometry (LSC) may aid in determining the transplant quality of islets...
  8. Goodale D, Phay C, Postenka C, Keeney M, Allan A. Characterization of tumor cell dissemination patterns in preclinical models of cancer metastasis using flow cytometry and laser scanning cytometry. Cytometry A. 2009;75:344-55 pubmed publisher
    ..of tumor dissemination to bone marrow (BM) and lymph node (LN), and for analysis of these samples by laser scanning cytometry (LSC)...
  9. Hamamoto T, Suzuki K, Kodama S, Sasaki H, Abe K, Hayashi T, et al. Correlation of malignant phenotypes of human tumour cell lines with augmented expression of Hsp72 protein measured by laser scanning cytometry. Int J Hyperthermia. 2007;23:363-70 pubmed
    ..880). These results indicate that LSC measurement is useful for predicting the degree of malignancy of cancer cells, as it is reliable, faster than Western blotting and more objective and quantitative than visual measurements. ..

More Information

Publications62

  1. Zhao H, Traganos F, Darzynkiewicz Z. Kinetics of the UV-induced DNA damage response in relation to cell cycle phase. Correlation with DNA replication. Cytometry A. 2010;77:285-93 pubmed publisher
    ..Phosphorylation of these proteins detected with phospho-specific Abs and measured by laser scanning cytometry in relation the cell cycle phase was found to be selective to S-phase cells...
  2. Darzynkiewicz Z, Smolewski P, Holden E, Luther E, Henriksen M, François M, et al. Laser scanning cytometry for automation of the micronucleus assay. Mutagenesis. 2011;26:153-61 pubmed publisher
    b>Laser scanning cytometry (LSC) provides a novel approach for automated scoring of micronuclei (MN) in different types of mammalian cells, serving as a biomarker of genotoxicity and mutagenicity...
  3. Pruimboom Brees I, Brees D, Shen A, Keener M, Francone O, Amacher D, et al. Using laser scanning cytometry to measure PPAR-mediated peroxisome proliferation and beta oxidation. Toxicol Pathol. 2005;33:86-91 pubmed
    b>Laser scanning cytometry (LSC) is a new technology that combines the properties and advantages of flow cytometry (FC) and immunohistochemistry (IHC), thus providing qualitative and quantitative information on protein expression with the ..
  4. Henriksen M, Miller B, Newmark J, Al Kofahi Y, Holden E. Laser scanning cytometry and its applications: a pioneering technology in the field of quantitative imaging cytometry. Methods Cell Biol. 2011;102:161-205 pubmed publisher
    ..b>Laser scanning cytometry (LSC), the original imaging cytometry technology, is an attractive choice for analysis of both cellular ..
  5. McGrath M, Morton A, Harnett M. Laser scanning cytometry: capturing the immune system in situ. Methods Cell Biol. 2011;102:231-60 pubmed publisher
    ..detect signaling events occurring in individual cells in situ and the development of technologies such as laser scanning cytometry that provide the tools to analyze physiologically relevant interactions between molecules and cells of ..
  6. Leifert W, François M, Thomas P, Luther E, Holden E, Fenech M. Automation of the buccal micronucleus cytome assay using laser scanning cytometry. Methods Cell Biol. 2011;102:321-39 pubmed publisher
    b>Laser scanning cytometry (LSC) can be used to quantify the fluorescence intensity or laser light loss (absorbance) of localized molecular targets within nuclear and cytoplasmic structures of cells while maintaining the morphological ..
  7. Darzynkiewicz Z, Halicka H, Zhao H. Analysis of cellular DNA content by flow and laser scanning cytometry. Adv Exp Med Biol. 2010;676:137-47 pubmed
    ..Described also are controls used to ensure accuracy of DNA ploidy determination, the principles in ploidy assessment and possible pitfalls in analysis. ..
  8. Nehmann N, Wicklein D, Schumacher U, Muller R. Comparison of two techniques for the screening of human tumor cells in mouse blood: quantitative real-time polymerase chain reaction (qRT-PCR) versus laser scanning cytometry (LSC). Acta Histochem. 2010;112:489-96 pubmed publisher
    ..Two techniques, namely quantitative real-time polymerase chain reaction (qRT-PCR) and laser scanning cytometry (LSC), were compared for screening of 100 microl blood samples from immunodeficient mice spiked with a ..
  9. Zhao H, Halicka H, Traganos F, Jorgensen E, Darzynkiewicz Z. New biomarkers probing depth of cell senescence assessed by laser scanning cytometry. Cytometry A. 2010;77:999-1007 pubmed publisher
  10. Stefan T, Jacobberger J. Laser scanning cytometry of mitosis: state and stage analysis. Methods Cell Biol. 2011;102:341-72 pubmed publisher
    ..We describe detection of cell states with quantitative immunofluorescence measurements performed by laser scanning cytometry. This platform allows both measurement of the cell states, capture of cell images within those states, ..
  11. Halicka H, Zhao H, Li J, Lee Y, Hsieh T, Wu J, et al. Potential anti-aging agents suppress the level of constitutive mTOR- and DNA damage- signaling. Aging (Albany NY). 2012;4:952-65 pubmed
    ..The data suggest that combined assessment of constitutive ?H2AX expression, mitochondrial activity (ROS, ??m) and mTOR signaling provides an adequate gamut of cell responses to evaluate effectiveness of gero-suppressive agents. ..
  12. Szodoray P, Jellestad S, Alex P, Zhou T, Wilson P, Centola M, et al. Programmed cell death of peripheral blood B cells determined by laser scanning cytometry in Sjögren's syndrome with a special emphasis on BAFF. J Clin Immunol. 2004;24:600-11 pubmed
    ..The B-cell apoptosis defects, the increased BAFF levels-correlating with hypergammaglobulinaemia-together with the raised B-cell cytokine levels indicates the disturbed B-cell biology in the disease. ..
  13. Hua W, Sheff D, Toomre D, Mellman I. Vectorial insertion of apical and basolateral membrane proteins in polarized epithelial cells revealed by quantitative 3D live cell imaging. J Cell Biol. 2006;172:1035-44 pubmed
  14. Baudart J, Olaizola A, Coallier J, Gauthier V, Laurent P. Assessment of a new technique combining a viability test, whole-cell hybridization and laser-scanning cytometry for the direct counting of viable Enterobacteriaceae cells in drinking water. FEMS Microbiol Lett. 2005;243:405-9 pubmed
  15. Holz R, Axelrod D. Secretory granule behaviour adjacent to the plasma membrane before and during exocytosis: total internal reflection fluorescence microscopy studies. Acta Physiol (Oxf). 2008;192:303-7 pubmed
    ..Increased travel may increase the probability of granules interacting productively with the plasma membrane constituents, thereby, increasing the probability of fusion. ..
  16. Bingham B, Kotnis S, McHendry Rinde B, Shen R, Wood A, Kennedy J. Laser scanning cytometry in the characterization of the proapoptotic effects of transiently transfected genes in cerebellar granule neurons. Cytometry A. 2006;69:1114-22 pubmed
    ..b>Laser scanning cytometry (LSC), with its high capacity, medium throughput means of collecting fluorescent emissions from cultured ..
  17. Ares I, Louzao M, Vieytes M, Yasumoto T, Botana L. Actin cytoskeleton of rabbit intestinal cells is a target for potent marine phycotoxins. J Exp Biol. 2005;208:4345-54 pubmed
    ..Those results identify the cytoskeleton as an early target for the toxic effect of those toxins. ..
  18. Kammerer B, Sardella B, Kültz D. Salinity stress results in rapid cell cycle changes of tilapia (Oreochromis mossambicus) gill epithelial cells. J Exp Zool A Ecol Genet Physiol. 2009;311:80-90 pubmed publisher
    ..mitochondria-rich cells (MRC) and other gill cells (non-MRC) within different cell cycle phases by laser scanning cytometry. Gill cells fixed on coverslips were triple stained with propidium iodide to distinguish G1 vs...
  19. Chakraborty A, Tansey W. Inference of cell cycle-dependent proteolysis by laser scanning cytometry. Exp Cell Res. 2009;315:1772-8 pubmed publisher
    ..We describe here the use of laser scanning cytometry (LSC), a technical merger between fluorescence microscopy and flow cytometry, to determine cell cycle-..
  20. Huss C, Krause M, Löw U, Riemann I, Stracke F, Mestres P, et al. [Experimental corneal imaging and corneal surgery with non-amplified femtosecond laser pulses]. Ophthalmologe. 2012;109:995-1000 pubmed publisher
    ..Easy switching between tissue imaging and ablation seems to be suitable for diagnostic and therapeutic applications. ..
  21. Zhao H, Li J, Traganos F, Halicka H, Zarebski M, Dobrucki J, et al. Cell fixation in zinc salt solution is compatible with DNA damage response detection by phospho-specific antibodies. Cytometry A. 2011;79:470-6 pubmed publisher
    ..and of ATM on Ser1981 was detected with phospho-specific Abs; cellular fluorescence was measured by laser scanning cytometry (LSC)...
  22. Juan G, Zoog S, Ferbas J. Leveraging image cytometry for the development of clinically feasible biomarkers: evaluation of activated caspase-3 in fine needle aspirate biopsies. Methods Cell Biol. 2011;102:309-20 pubmed publisher
    Quantitation of activated caspases in xenograft models by laser scanning cytometry has demonstrated mechanism-specific biological activity of Anti-Trail Receptor immunoglobulin therapies in situ...
  23. Guarguaglini G, Duncan P, Stierhof Y, Holmström T, Duensing S, Nigg E. The forkhead-associated domain protein Cep170 interacts with Polo-like kinase 1 and serves as a marker for mature centrioles. Mol Biol Cell. 2005;16:1095-107 pubmed
    ..We show that Cep170 labeling can be used to discriminate bona fide centriole overduplication from centriole amplification that results from aborted cell division. ..
  24. Awasthi S, Matthews D, Li R, Chiamvimonvat N, Lieu D, Chan J. Label-free identification and characterization of human pluripotent stem cell-derived cardiomyocytes using second harmonic generation (SHG) microscopy. J Biophotonics. 2012;5:57-66 pubmed publisher
    ..Our study demonstrates the feasibility of developing a SHG-activated flow cytometer for the non-invasive purification of PSC-CMs. ..
  25. Savatier J, Gbankoto A, Vigo J, Salmon J. Videomicrofluorometry on living cells and discriminant factorial analysis to study cell cycle distributions. J Biol Regul Homeost Agents. 2004;18:206-11 pubmed
    ..Such an approach allows to accurately evidence the change of the values of some cellular parameters. ..
  26. Rosner K, Kasprzak M, Horenstein A, Thurston H, Abrams J, Kerwin L, et al. Engineering a waste management enzyme to overcome cancer resistance to apoptosis: adding DNase1 to the anti-cancer toolbox. Cancer Gene Ther. 2011;18:346-57 pubmed publisher
    ..Quantification of apoptosis by Laser scanning cytometry demonstrated high-killing efficiency of 70-100%...
  27. Greene C, Ramsay H, Wells R, O Neill S, McElvaney N. Inhibition of Toll-like receptor 2-mediated interleukin-8 production in Cystic Fibrosis airway epithelial cells via the alpha7-nicotinic acetylcholine receptor. Mediators Inflamm. 2010;2010:423241 pubmed publisher
  28. Gerstner A, Mittag A, Laffers W, Dähnert I, Lenz D, Bootz F, et al. Comparison of immunophenotyping by slide-based cytometry and by flow cytometry. J Immunol Methods. 2006;311:130-8 pubmed
    ..The detection of rare leukocytes or weak antigens is limited; in these cases appropriate amplification steps for immunofluorescence should be engaged. ..
  29. Bowen W, Wylie P. Application of laser-scanning fluorescence microplate cytometry in high content screening. Assay Drug Dev Technol. 2006;4:209-21 pubmed
    ..The review focuses on the Acumen Explorer microplate cytometer (TTP LabTech Ltd., Melbourn, Hertfordshire, UK), detailing the principal components of the instrument and providing an overview of its use in high content screening. ..
  30. Szaflik J. Comparison of in vivo confocal microscopy of human cornea by white light scanning slit and laser scanning systems. Cornea. 2007;26:438-45 pubmed
  31. Holme A, Yadav S, Pervaiz S. Automated laser scanning cytometry: a powerful tool for multi-parameter analysis of drug-induced apoptosis. Cytometry A. 2007;71:80-6 pubmed
    ..Here we used the automated laser scanning cytometry (LSC) for multi-parameter analysis of drug-induced tumor cell apoptosis...
  32. Roesch Ely M, Steinberg T, Bosch F, Mussig E, Whitaker N, Wiest T, et al. Organotypic co-cultures allow for immortalized human gingival keratinocytes to reconstitute a gingival epithelial phenotype in vitro. Differentiation. 2006;74:622-37 pubmed
    ..We are not aware of any other OCC system comprising of IHGK, which allows for site-specific expression of gingival epithelial markers. This substantiates reconstitution of a gingival epithelial phenotype in vitro. ..
  33. Mansfield J, Hoyt C, Levenson R. Visualization of microscopy-based spectral imaging data from multi-label tissue sections. Curr Protoc Mol Biol. 2008;Chapter 14:Unit 14.19 pubmed publisher
    ..These commercially available tools can be attached to the standard laboratory light microscope to significantly enhance the power of light microscopy. ..
  34. Mital J, Schwarz J, Taatjes D, Ward G. Laser scanning cytometer-based assays for measuring host cell attachment and invasion by the human pathogen Toxoplasma gondii. Cytometry A. 2006;69:13-9 pubmed
    ..The LSC-based assays represent useful new methods for quantitatively measuring attachment and invasion by T. gondii, and can be readily adapted to study similar processes in other host-pathogen systems. ..
  35. Zoog S, Itano A, Trueblood E, Pacheco E, Zhou L, Zhang X, et al. Antagonists of CD117 (cKit) signaling inhibit mast cell accumulation in healing skin wounds. Cytometry A. 2009;75:189-98 pubmed publisher
    ..Here we describe an automated laser scanning cytometry assay that was used to characterize the kinetics of MC accumulation in healing skin wounds and to study ..
  36. Abdelrahim M, Baker C, Abbruzzese J, Safe S. Tolfenamic acid and pancreatic cancer growth, angiogenesis, and Sp protein degradation. J Natl Cancer Inst. 2006;98:855-68 pubmed
    ..009) than tumors from control mice. Tolfenamic acid is a new antipancreatic cancer NSAID that activates degradation of transcription factors Sp1, Sp3, and Sp4; reduces VEGF expression; and decreases tumor growth and metastasis. ..
  37. Glauner H, Ruttekolk I, Hansen K, Steemers B, Chung Y, Becker F, et al. Simultaneous detection of intracellular target and off-target binding of small molecule cancer drugs at nanomolar concentrations. Br J Pharmacol. 2010;160:958-70 pubmed publisher
    ..The combination of FCS and FCCS provides a powerful means to assess subcellular pharmacokinetics and dynamics of drug candidates at nanomolar concentrations. ..
  38. Sun L, Li D, Liu Z, Yang L, Liu J, Cao J. Analysis of micronuclei in the transferrin-receptor positive reticulocytes from peripheral blood of nasopharyngeal cancer patients undergoing radiotherapy by a single-laser flow cytometer. J Radiat Res. 2005;46:25-35 pubmed
    ..Our results indicated that the single-laser flow cytometric system was quick, reasonable and acceptable for detecting the frequency of micronucleated transferrin-receptor positive reticulocytes from human peripheral blood. ..
  39. Gajewska M, Gajkowska B, Motyl T. Apoptosis and autophagy induced by TGF-B1 in bovine mammary epithelial BME-UV1 cells. J Physiol Pharmacol. 2005;56 Suppl 3:143-57 pubmed
    ..b>Laser scanning cytometry and confocal microscopy were used for analysis of MAP1 LC3 and Beclin 1 expression - two proteins ..
  40. Shao L, Wood C, Zhang D, Tannir N, Matin S, Dinney C, et al. Telomere dysfunction in peripheral lymphocytes as a potential predisposition factor for renal cancer. J Urol. 2007;178:1492-6 pubmed
    Genetic integrity is maintained in part by the architecture of telomeres. We previously developed a laser scanning cytometry based quantitative fluorescence in situ hybridization assay to assess telomere length in peripheral blood ..
  41. Aguila B, Roussel M, Jauzac P, Allouche S. High-purity selection and maintenance of gene expression in human neuroblastoma cells stably over-expressing GFP fusion protein. Application for opioid receptors desensitization studies. Brain Res. 2006;1114:11-8 pubmed
    ..Thus, this approach provides a valuable tool for exploring opioid receptors desensitization in neuronal cells. ..
  42. Herbst R. Imaging in drug development. Clin Adv Hematol Oncol. 2004;2:268-9 pubmed
  43. Kümmel A, Selzer P, Siebert D, Schmidt I, Reinhardt J, Götte M, et al. Differentiation and visualization of diverse cellular phenotypic responses in primary high-content screening. J Biomol Screen. 2012;17:843-9 pubmed publisher
    ..The approach has the potential to identify both expected hits and novel cellular phenotypes. Moreover, different chemotypes, which cause the same phenotypic effects, are identified, thus facilitating "scaffold hopping." ..
  44. Gerster A, Trumpfheller C, Racz P, Osmancik P, Tenner Racz K, Tarnok A. Quantitative histology by multicolor slide-based cytometry. Cytometry A. 2004;61:210-9 pubmed
    ..We have developed a semiautomated analysis method for laser scanning cytometry (LSC) termed "multiple thresholding," which is suitable for archived or fresh biopsy material ..
  45. Al Rawi M, Diabaté S, Weiss C. Uptake and intracellular localization of submicron and nano-sized SiO? particles in HeLa cells. Arch Toxicol. 2011;85:813-26 pubmed publisher
    ..No accumulation of SiO(2) particles in nuclei or mitochondria of HeLa cells could be observed. In contrast to SiO(2)-SMPs, SiO(2)-NPs are preferentially localized in lysosomes. ..
  46. Lin J, Della Fera M, Baile C. Green tea polyphenol epigallocatechin gallate inhibits adipogenesis and induces apoptosis in 3T3-L1 adipocytes. Obes Res. 2005;13:982-90 pubmed
    ..for 24 hours with 0 to 200 microM EGCG, then stained with annexin V and propidium iodide and analyzed by laser scanning cytometry. Both preadipocytes and adipocytes were also analyzed for apoptosis by terminal deoxynucleotidyl ..
  47. Mach W, Thimmesch A, Orr J, Slusser J, Pierce J. Flow cytometry and laser scanning cytometry, a comparison of techniques. J Clin Monit Comput. 2010;24:251-9 pubmed publisher
    Flow and laser scanning cytometry are used extensively in research and clinical settings. These techniques provide clinicians and scientists information about cell functioning in a variety of health and disease states...
  48. Gerstner A, Tarnok A. Analysis of fine-needle aspirate biopsies from solid tumors by laser scanning cytometry (LSC). Curr Protoc Cytom. 2002;Chapter 7:Unit 7.20 pubmed publisher
    ..Tumors located at sites unsuitable for routine biopsy can be analyzed even on an out-patient basis. ..
  49. Leavesley H, Li L, Prabhakaran K, Borowitz J, Isom G. Interaction of cyanide and nitric oxide with cytochrome c oxidase: implications for acute cyanide toxicity. Toxicol Sci. 2008;101:101-11 pubmed
    ..b>Laser scanning cytometry showed that cyanide elevated mitochondrial NO, which then was available to interact with CcOX to enhance ..
  50. Ichii H, Inverardi L, Pileggi A, Molano R, Cabrera O, Caicedo A, et al. A novel method for the assessment of cellular composition and beta-cell viability in human islet preparations. Am J Transplant. 2005;5:1635-45 pubmed
    ..of beta-cell content and viability in human islets based on cellular composition analysis through laser scanning cytometry (LSC) coupled with identification of beta-cell-specific apoptosis at the mitochondrial level...
  51. Sanislo L, Vertaková Krakovská B, Kuliffay P, Brtko J, Galbava A, Galbavy S. Detection of circulating tumor cells in metastatic breast cancer patients. Endocr Regul. 2011;45:113-24 pubmed
    ..This was followed by analysis of isolated cells with the aid of laser scanning cytometry (LSC)...
  52. Kornblau S, Qiu Y, Bekele B, Cade J, Zhou X, Harris D, et al. Studying the right cell in acute myelogenous leukemia: dynamic changes of apoptosis and signal transduction pathway protein expression in chemotherapy resistant ex-vivo selected "survivor cells". Cell Cycle. 2006;5:2769-77 pubmed
    ..Expression of BCL2, bax, PKC alpha, ERK2 and pERK2 proteins was measured using laser scanning cytometry. The SV population was enriched for CD34+ stem cells...
  53. King J, Pink M, Jobe C. Assessment of reprocessed arthroscopic shaver blades. Arthroscopy. 2006;22:1046-52 pubmed
    ..to cut meniscal tissue, and the cut surface was measured for smoothness by image processing and automated laser scanning cytometry. In evaluation of the meniscus, for the subset of shavers, an image processing value of 1 indicates a ..