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The Correlation Confocal Microscope  [PDF]
D. S. Simon,A. V. Sergienko
Physics , 2010, DOI: 10.1364/OE.18.009765
Abstract: A new type of confocal microscope is described which makes use of intensity correlations between spatially correlated beams of light. It is shown that this apparatus leads to significantly improved transverse resolution.
Detection of Intra-Tumor Self Antigen Recognition during Melanoma Tumor Progression in Mice Using Advanced Multimode Confocal/Two Photon Microscope  [PDF]
David A. Schaer, Yongbiao Li, Taha Merghoub, Gabrielle A. Rizzuto, Amos Shemesh, Adam D. Cohen, Yanyun Li, Francesca Avogadri, Ricardo Toledo-Crow, Alan N. Houghton, Jedd D. Wolchok
PLOS ONE , 2011, DOI: 10.1371/journal.pone.0021214
Abstract: Determining how tumor immunity is regulated requires understanding the extent to which the anti-tumor immune response “functions” in vivo without therapeutic intervention. To better understand this question, we developed advanced multimodal reflectance confocal/two photon fluorescence intra-vital imaging techniques to use in combination with traditional ex vivo analysis of tumor specific T cells. By transferring small numbers of melanoma-specific CD8+ T cells (Pmel-1), in an attempt to mimic physiologic conditions, we found that B16 tumor growth alone was sufficient to induce naive Pmel-1 T cell proliferation and acquisition of effector phenotype. Tumor -primed Pmel-1 T cells, are capable of killing target cells in the periphery and secrete IFNγ, but are unable to mediate tumor regression. Within the tumor, Pmel-1 T cells have highly confined mobility, displaying long term interactions with tumor cells. In contrast, adoptively transferred non tumor-specific OT-I T cells show neither confined mobility, nor long term interaction with B16 tumor cells, suggesting that intra-tumor recognition of cognate self antigen by Pmel-1 T cells occurs during tumor growth. Together, these data indicate that lack of anti-tumor efficacy is not solely due to ignorance of self antigen in the tumor microenvironment but rather to active immunosuppressive influences preventing a protective immune response.
ConfocalCheck - A Software Tool for the Automated Monitoring of Confocal Microscope Performance  [PDF]
Keng Imm Hng, Dirk Dormann
PLOS ONE , 2013, DOI: 10.1371/journal.pone.0079879
Abstract: Laser scanning confocal microscopy has become an invaluable tool in biomedical research but regular quality testing is vital to maintain the system’s performance for diagnostic and research purposes. Although many methods have been devised over the years to characterise specific aspects of a confocal microscope like measuring the optical point spread function or the field illumination, only very few analysis tools are available. Our aim was to develop a comprehensive quality assurance framework ranging from image acquisition to automated analysis and documentation. We created standardised test data to assess the performance of the lasers, the objective lenses and other key components required for optimum confocal operation. The ConfocalCheck software presented here analyses the data fully automatically. It creates numerous visual outputs indicating potential issues requiring further investigation. By storing results in a web browser compatible file format the software greatly simplifies record keeping allowing the operator to quickly compare old and new data and to spot developing trends. We demonstrate that the systematic monitoring of confocal performance is essential in a core facility environment and how the quantitative measurements obtained can be used for the detailed characterisation of system components as well as for comparisons across multiple instruments.
Supercontinuum ultra wide range confocal microscope for reflectance spectroscopy of living matter and material science surfaces
Stefano Selci,Francesca R. Bertani,Luisa Ferrari
AIP Advances , 2011, DOI: 10.1063/1.3631661
Abstract: We report the design and implementation of a new reflectance laser scanning confocal system with spectroscopy imaging capabilities. Confocal spectroscopy is achieved by using a very broad spectral range supercontinuum source capable of high precision reflectance data in the VIS-IR spectral range thanks to an almost achromatic optical layout. With this apparatus we collect each single scanning point as a whole spectrum in a continuous range, associated with the optical section imaging possibilities typical of a confocal set up. While such a microscope has been developed for bio medical analysis of human skin and other similar applications, first test results on solid samples produce spectroscopic results that, compared to analytical models based on the Abelés matrix transfer methods, show a very good agreement, opening new possibilities of a complete spectroscopic fingerprinting of samples with microscopic details.
Living Matter Observations with a Novel Hyperspectral Supercontinuum Confocal Microscope for VIS to Near-IR Reflectance Spectroscopy  [PDF]
Francesca R. Bertani,Luisa Ferrari,Valentina Mussi,Elisabetta Botti,Antonio Costanzo,Stefano Selci
Sensors , 2013, DOI: 10.3390/s131114523
Abstract: A broad range hyper-spectroscopic microscope fed by a supercontinuum laser source and equipped with an almost achromatic optical layout is illustrated with detailed explanations of the design, implementation and data. The real novelty of this instrument, a confocal spectroscopic microscope capable of recording high resolution reflectance data in the VIS-IR spectral range from about 500 nm to 2.5 μm wavelengths, is the possibility of acquiring spectral data at every physical point as defined by lateral coordinates, X and Y, as well as at a depth coordinate, Z, as obtained by the confocal optical sectioning advantage. With this apparatus we collect each single scanning point as a whole spectrum by combining two linear spectral detector arrays, one CCD for the visible range, and one InGaAs infrared array, simultaneously available at the sensor output channel of the home made instrument. This microscope has been developed for biomedical analysis of human skin and other similar applications. Results are shown illustrating the technical performances of the instrument and the capability in extracting information about the composition and the structure of different parts or compartments in biological samples as well as in solid statematter. A complete spectroscopic fingerprinting of samples at microscopic level is shown possible by using statistical analysis on raw data or analytical reflectance models based on Abelés matrix transfer methods.
Using laser confocal scanning microscope to study ischemia-hypoxia injury in rat brain slice
Xiaoying Wang,Hong Xing,Qihua He,Jialing Xu,Benjie Wu
Chinese Science Bulletin , 2000, DOI: 10.1007/BF02884902
Abstract: The level of lipid peroxidation and cellular necrosis in rat living brain slices during brain ischemia-hypoxia injury have been observed using a laser confocal scanning microscope (LCSM) with double labeling of fluorescent probes D-399 (2, 7-dichlorofluorescin diacetate) and propidium iodide (PI). The hypoxia and/or reoxygenation injury in rat brain slices is markedly decreased by pretreatment with L-NG-nitro-arginine (L-NNA) and N-acetylcysteine (NAC), showing that the nitric oxide (NO) and other free radicals play an important role in brain ischemia-hypoxia injury.
Using laser confocal scanning microscope to study ischemia-hypoxia injury in rat brain slice
WANG Xiaoying,

科学通报(英文版) , 2000,
Abstract: The level of lipid peroxidation and cellular necrosis in rat living brain slices during brain ischemia-hypoxia injury have been observed using a laser confocal scanning microscope (LCSM) with double labeling of fluorescent probes D-399 (2, 7-dichlorofluorescin diacetate) and propidium iodide (PI). The hypoxia and/or reoxygenation injury in rat brain slices is markedly decreased by pretreatment with L-NG-nitro-arginine (L-NNA) and N-acetylcysteine (NAC), showing that the nitric oxide (NO) and other free radicals play an important role in brain ischemia-hypoxia injury.
Effects of Naphthalene on DNA and RNA quantity in Amoeba proteus by using confocal laser scanning microscope
Khwanmuni, J.,Tansakul, R.
Songklanakarin Journal of Science and Technology , 2006,
Abstract: Effects of Naphthalene which is a carcinogen on changes of DNA and RNA quantities were studied with acridine orange stained cells under a confocal laser scanning microscope. It was found that DNA and RNA in amoebae nucleus and cytoplasm, reared in 0 (control), 3 and 8.85 mg/l (24h-LD50) at 0 and 12 h. showed a statistically significant difference (p<0.05). The more naphthalene concentrations and larger incubation periods had greater effects on DNA and RNA decreases in amoebae nucleus and cytoplasm.
Qualitative analysis of re mineralized carious lesions subjected to fluoride supplement through confocal laser scanning microscope  [PDF]
K. Shashikala, N. V. Sheela
Open Journal of Stomatology (OJST) , 2011, DOI: 10.4236/ojst.2011.13010
Abstract: Aim: 1] Comparative evaluation of the linear depth of induced remineralized lesions after subjecting to fluoride supplements and 2] To assess the average fluorescence at both the demineralized and the remi-neralized zones in all the three study groups under confocal laser scanning microscope. Method: Forty five sound human premolars extracted for orthodon-tic reasons were decoronated 1 mm below the ce-mento-enamel junction and coated with nail varnish except for a 3 × 3 mm window on the buccal surface. The samples were placed in 50 ml of de mineralizing solution at pH 4.6 for 96 hours. Following deminera-lization, the lower half of the 3 × 3 mm window in all the samples were covered with nail varnish to serve as control. The samples were randomly divided into three groups of fifteen teeth each (n = 15) and speci-mens in group A[Nfd] were remineralized using non-fluoridated dentifrice [control], those in groups B [Fd5] and group C [Fd10] using 500 ppm and 1000 ppm of fluoride containing dentifrice, respectively. The specimens were subjected to a 20 day reminera-lization treatment regimen and were sectioned into 100 µm thick sections and two images were captured on the buccal surface from either side of the midpoint of occluso-cervical length using confocal laser scan-ning microscope [CLSM]. Results: were tabulated and statistically analyzed by Anova. Study concluded that 1000 ppm fluoridated dentifrice showed a greater degree of remineralization than other groups and confocal laser scanning microscopes gives promising results in the diagnosis of early enamel lesions over the conventional methods.
Quantitative read-out of Al2O3:C,Mg-based fluorescent nuclear track detectors using a commercial confocal microscope  [PDF]
Steffen Greilich,Julia-Maria Osinga,Martin Niklas,Florian Lauer,Felix Bestvater,Oliver J?kel
Physics , 2014,
Abstract: Fluorescent nuclear track detectors (FNTD) show great potential for applications in ion-beam therapy research, such as dosimetry, advanced beam characterization, in-vivo use or as radiobiological assay. A essential feature of FNTDs is their ability to assess the energy loss of single ions yielding for example LET estimations. This article describes the basic characterisations of FNTDs and our read-out system (a Zeiss LSM710 confocal laser scanning microscope) to enable quantative measurements of energy loss.
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