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

▶
cellular imaging core
SMART_READER_LITE
LIVE PREVIEW

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

Jul 10, 2023 7 likes •204 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

slide-1
SLIDE 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

slide-2
SLIDE 2

Image Acquisition and storage Image analysis

5 10 15 20 25 30 35 40 10 20 30 40 50 60 70 Time (10 second intervals) Average Intensity

1 2 4 3 5

5 10 15 20 25 30 35 40 10 20 30 40 50 60 70 Time (10 second intervals) Average Intensity

1 2 4 3 5

Image processing

A B C

slide-3
SLIDE 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
slide-4
SLIDE 4

Technologies first available at BU/BMC

slide-5
SLIDE 5

Cyntellect

LEAP ( Laser-Enabled Analysis and Processing)

slide-6
SLIDE 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.

slide-7
SLIDE 7

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%.

Cyntellect: Cell enrichment by laser based elimination

slide-8
SLIDE 8

Automated in situ purification of primary rat brain microvascular endothelial cells

before after

slide-9
SLIDE 9

Cell Monolayer Wounds

slide-10
SLIDE 10

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

slide-11
SLIDE 11

Cloning of a hyper-secreting cell

slide-12
SLIDE 12

Zeiss LSM 710 NLO LIVE DUO

slide-13
SLIDE 13

Carl Zeiss Inc.

LSM 710

slide-14
SLIDE 14

Carl Zeiss Inc.

Live5

slide-15
SLIDE 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
slide-16
SLIDE 16

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

Live Cell Array (Molecular Cytomics)

slide-17
SLIDE 17

1 : GFP positive cell, didn’t differentiate, no benzidine 2: GFP positive cell, did differentiate, benzidine 3: GFP negative cell, did differentiate, benzidine

GFP GlyA Merge Bright field after benzidine application

slide-18
SLIDE 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