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    Kansas State University Microscopy Facility
    Division of Biology
    Ackert Hall

    1) Services and Equipment Available
    2) Schedule of Charges
    3) Classes Offered
    4) Images
    5) Publications

    Services and Equipment Available

    Complete individual or small group training and familiarization with equipment and techniques for transmission electron, fluorescence and confocal microscopies. Clients can be trained in specimen preparation and use of instruments themselves or may request assistance from the facility's staff.

    Collaborative efforts in experimental design, data analysis and interpretation, manuscript and grant preparation for researchers seeking to use microscopy to address research questions.

    FEI CM100 Transmission Electron Microscope equipped with an AMT digital image capturing system, negative film image capturing system, accelerating voltages up to 100kV, low-dose capabilities, electron diffraction, a CompuStage capable of storing 25 x,y and z locations on a grid and tilting up to 65 degree, a cold-finger, a Gatan cryo-holder and Ethernet connection.

     

    Zeiss Axioplan 2  equipped with a Zeiss Axiocam HR digital camera, a fully motorized stage with mark and find software, Plan Neofluor objectives (1.25x/0.035, 10x/0.3, 20x/0.5, 40x/0.75, 40x/1.3 oil), Plan apochromat objectives (63x/1.4 oil, 100x/1.4 oil), differential contrast interference (DIC), phase contrast (ph), HBO burner, dark field, bright field, specific filter sets for proper excitation and observation of blue, green, yellow and red fluorescence, and Axiovision 3.1 software with interactive measurements and 3D deconvolution modules. This microscope is currently capable of recording multichannel- fluorescence, z-stacks over time from multiple locations on a sample. The HBO light source is capable of exciting dyes in the range from 350-700 nm.
     


    New Zeiss LSM 5 PASCAL
    (
    Laser Scanning Confocal Microscope). This system is capable of exciting fluorescent markers with laser lines of 458/488/514/543/633 nm, observation of blue, green, yellow, orange and red fluorescence, single and simultaneous double fluorescence imaging, multitracking of multifluorescence signals, acquisition of image stacks, 3D visualization, time series, multiple time series, FRET, quantitative colocalization, orthogonal slicing of 3D data, and visualization and analysis of ion concentrations.



    Complete tissue/sample preparation facilities including ultramicrotomes, stereo microscopes, Kinney vacuum evaporator and more.

    Two complete darkroom suites equipped for film development and for enlargement of images onto photographic paper.

    Complete Image processing and analysis facilities equipped with PC work stations equipped with image analysis and processing software (Adobe Photoshop, Scion Image, ImageJ, Omni Page Pro, MS Power Point, Sigmaplot, SlideWrite, FTP), flat bed scanners for digitizing images and text, Polaroid Digital Palettes for creating slides, a Lexmark Optra R+ laser printer, Mitsubishi CP210U dye-sublimation printer for photographic quality prints and Ethernet connections.

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    Schedule of Charges 

    The Division of Biology is authorized to make the following charges for services rendered and materials and supplies furnished:

    Description of Services and Materials

    Charges per hour

    1. Processing of samples, individual training in microscopy and/or technical consultation

    $25.00

    2. Axioplan 2

    $10.00

    3. Beam time on FEI CM 100 Transmission Electron Microscope (per beam hour used).

    $55.00

    4. Beam time on Zeiss Laser Scan Confocal Microscope (LSM).

    $20.00

    5. Supplies provided by the facility for sample preparation, photograph and imaging.

    Cost plus 10% handling



    We request that all Facility users include a statement of acknowledgment in their research publications.  A sample statement would be: 

    We acknowledge the Kansas State University Biology Microscopy  Facility, which has been supported in part by  University resources, and  the Kansas Agricultural Experiment Station.

    For more information about this facility contact: Daniel L. Boyle, Director, Phone (785) 532-0134 (office) or 6647(lab), E-mail: dboyle@ksu.edu.

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    Classes Offered

    BIOL 886. Confocal, Fluorescence and Light Microscopy. (3) I, in odd years. An introduction to theories, functions and applications of confocal, fluorescence and light microscopy, and fluorescent molecules. Lab emphasis on students working on independent research projects requiring microscopy. Two hours of lecture and three hours of lab per week.

    BIOL 888. Electron Microscopy Techniques. (3) I, in even years. Theory and techniques involved in using the transmission electron microscope for the study of biological materials. Includes individualized instruction on the operation of transmission electron microscopes and techniques for processing biological samples. Pr.: Current participation in research requiring electron microscope.

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    Images













    Negative stained viruses



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    Publications

    1) Boyle, D. and L. Takemoto (1994) Characterization of the a-g and a-b complex:Evidence for an in vivo functional role of a-crystallin as a molecular chaperone. Exp. Eye Res. 58:9-16.

    2) Takemoto, L. and D. Boyle (1994) Molecular chaperone properties of the high molecular weight aggregate from aged lens. Curr. Eye Res. 13:35-44.

    3) Boyle, D. and L. Takemoto (1994) Immunolocalization of the C-terminal and N-terminal region of alpha-A and alpha-B crystallins. Curr. Eye Res. 13:497-504.

    4) Takemoto, L. and D. Boyle (1994) Binging of denatured protein decreases the chaperone properties of alpha crystallin. Arch. Biochem. Biophys. 315:133-136.

    5) Yan, F.,D.A. Lutz, V.L. Shepherd, D. Boyle, and B.J. McLaughlin (1995) Characterization of rod outer segment plasma membrane proteins which bind to the mannose receptor. Curr. Eye Res. 14:465-471.

    6) Boyle, D. and L. Takemoto (1995) EM immunolocalization of a-crystallins: Association with the plasma membrane from normal and cataract human lenses. Curr. Eye Res. 15:577-582.

    7) Boyle, D. L., D. S. Blunt and L. J. Takemoto (1997) Confocal microscopy of cataracts from animal model systems : Relevance to human nuclear cataract. Exp. Eye Res. 64;565-572.

    8) Boyle, D. L. and L. J. Takemoto (1997) Laser scanning confocal microscopy of human lens membranes in aged normal and nuclear cataracts. Invest. Ophthalmol. Vis. Sci. 38:2826-2832.

    9) Takemoto, L. and D. Boyle (1998) The possible role of alpha crystallins in human senile cataractogenesis. Inter. J. Biol. Macromol. 22:331-337.

    10) Takemoto, L. and D. Boyle (1998) Determination of the in vivo deamindation of asparagine-101 from alpha-A crystallin using microdissected sections of the aging human lens. Exp. Eye Res. 67:119-120.

    11) Takemoto, L. and D. Boyle (1998) Deamidation of specific glutamine residues from alpha-A crystallin during aging of the human lens. Biochem. 37:13681-13685.

    12) Boyle, D.L. and L. Takemoto (1998) Finger-like projections of plasma membrane in the most senescent fiber cells of human lenses. Curr. Eye Res. 17:1118-1123.

    13) Boyle, D.L. and L. J. Takemoto (1999) Localization of MIP 26 in nuclear fiber cells from aged normal and age-related nuclear cataractous human lenses. Exp. Eye Res. 68:41-49.

    14) Takemoto, L. and D. Boyle (1999) Deamidation of alpha-A crystallin from nuclei ofcataractous and normal human lenses. Mol. Vis. 5:2.

    15) DL Boyle and L.J. Takemoto (2000) A possible role for a-crystallins in lens epithelial cell differentiation.  Mol. Vis. 6:63-71.

    16) Y. Tamada, C.  Fukiage, D.L. Boyle, M. Azuma and T.R. Shearer (2000) Involvement of cysteine proteases in bFGF-induced angiogenesis in guinea pig and rat cornea. J. Ocular Pharmacol. 16:271-283.

    17) Wagner LM, Saleh SM, Boyle DL, Takemoto DJ. Effects of protein kinase Cγ on gap junction disassembly in lens epithelial cells and retinal cells in culture. Mol Vis. 2002; 8:59-66. http://www.molvis.org/molvis/v8/a8/

    18) Tasheva ES, Koester A , Paulsen AQ, Garrett AS, Boyle DL, Davidson HJ, Song M, Fox N, Conrad GW. Mimecan/osteoglycin-deficient mice have collagen fibril abnormalities. Mol Vis. 2002; 8:407-415.  http://www.molvis.org/molvis/v8/a48/ 

    19) Boyle DL, Carman P, Takemoto L. Translocation of macromolecules into whole rat lenses in culture. Mol Vis. 2002; 8:226-234. http://www.molvis.org/molvis/v8/a29/

    20) Nguyen TA, Boyle DL, Wagner LM, Shinohara T, Takemoto DJ. LEDGF active of PKCγ and gap junction disassembly in lens epithelial cells. Experimental Eye Res. 2003, 76(5):565-72.

    21) Lin D, Boyle DL, Takemoto DJ. IGF-1-induced phosphorylation of connexin 43 by PKC γ: Regulation of gap junctions in rabbit lens epithelial cells. Invest. Ophthalmol Vis. Sci. 2003, 44(3):1160-8.

    22) Boyle DL, Takemoto L, Brady JP, Wawrousek EF. Morphological characterization of the alphaA- and alphaB-crystallin double knockout mouse lens. BMC Ophthalmology. 2003, 3:3. [http://www.biomedcentral.com/1471-2415/3/3]

    23)  Tasheva ES, Ke An,  Boyle DL,  Conrad G W.. Expression and localization of leucine-rich B7 protein in human ocular tissues. Mol. Vis.  6 July 2005 [Full text]


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