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Cellular Imaging Core Orian Shirihai, MD PhD Scientific Director - PowerPoint PPT Presentation

Jul 10, 2023 •7 likes •187 views

Cellular Imaging Core Orian Shirihai, MD PhD Scientific Director shirihai@bu.edu Mike Kirber, PhD Technical Director Tel: 617-638-7153 Cellphone: 617-571-3408 mkirber@bu.edu Image Image processing B A Acquisition and storage 40 40

  1. Cellular Imaging Core Orian Shirihai, MD PhD Scientific Director shirihai@bu.edu Mike Kirber, PhD Technical Director Tel: 617-638-7153 Cellphone: 617-571-3408 mkirber@bu.edu

  2. Image Image processing B A Acquisition and storage 40 40 35 35 30 30 Average Intensity Average Intensity 1 1 25 25 20 20 4 4 15 15 3 3 5 5 10 10 2 2 5 5 0 0 0 0 10 10 20 20 30 30 40 40 50 50 60 60 70 70 Time (10 second intervals) Time (10 second intervals) Image analysis C

  3. Additional support for imaging studies • Incubators, hood, wet lab area for cultures and tissue specimens • File server for short and medium-term data storage • Workstations for image processing with high-speed link to server • Software support for image processing and analysis

  4. Technologies first available at BU/BMC

  5. Cyntellect LEAP ( Laser-Enabled Analysis and Processing)

  6. Cyntellect: Opto injection Optoinjection of Living Cells. LEAP™ employs targeted lasers to transiently permeabilize cells allowing uptake of a wide variety of molecules including certain: (a) ions, (b) small molecules, (c) dextran, (d) proteins, (e) fluorescent biosensors, and (f) QDots™ quantum dots.

  7. Cyntellect: Cell enrichment by laser based elimination A sample B cell population (green) contaminated with ~40% T cells tagged with a phycoerythrin-tagged T cell specific antibody (red) is imaged and analyzed by LEAP™ . Following laser processing of this same sample by LEAP™ , both resulting purity and yield exceeded 99%.

  8. Automated in situ purification of primary rat brain microvascular endothelial cells after before

  9. Cell Monolayer Wounds

  10. Development of Highly-Secreting Cell Lines Selection of highly secreting cells for further analysis

  11. Cloning of a hyper-secreting cell

  12. Zeiss LSM 710 NLO LIVE DUO

  13. LSM 710 Carl Zeiss Inc.

  14. Live5 Carl Zeiss Inc.

  15. Zeiss LSM 710 –Live5 DUO • Laser scanning confocal • Fast scanning by Live 5 (120 frames/sec) • 2-Photon guided by the LSM 710 scanner • 37C and CO2 control on microscope stage

  16. Live Cell Array (Molecular Cytomics) Live Cell Array • Monitor multiple living cells over days at the resolution of the individual cell • Fix cells in the array and determine expression of specific proteins

  17. GlyA GFP Bright field after Merge benzidine application 1 : GFP positive cell, didn’t differentiate, no benzidine 2: GFP positive cell, did differentiate, benzidine 3: GFP negative cell, did differentiate, benzidine

  18. Administrative •On Line Scheduling System-coming soon •Support letters for each core, please contact Maria LoSurdo at maria.losurdo@bmc.org or 617-638- 6957 •Any questions pertaining to billing, scheduling, please contact Maria LoSurdo

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