Tissue culture and molecular

The Tissue Culture and Molecular core provides visual science investigators (with priority for those with qualifying NEI R01 grants) with facilities and technical assistance for the maintenance and propagation of ocular and other cell types, as well as genotyping, RT and Q-PCR, Reactive Oxygen Species (ROS), and Adeno-Associated Virus vector production.

The broad functions of Tissue Culture Module

• isolation of primary cells from ocular tissue;
• maintenance and passage of established cell lines, including storage;
• large scale amplification of cell lines;
• training in growth and maintenance of cells as well as training on module equipment;
• strategic advice in planning experiments and determining their feasibility.
   

The core provides genotyping of mice, including cell-specific knockout mice. The core provides consultation/assistance on recombinant (r) AAV virus vector design and construction as well as assistance to the PI or staff for in vitro functional assessment. The core also assists large scale high quality AAV packaging through an external provider with discounted price. Equipment available for use in the Tissue Culture portion of the module includes:

• Azure 600
• CFX Opus 96 PCR machine
• Eppendorf 5804 benchtop centrifuge
• Two single chamber water jacketed CO2 incubator
• Olympus CK- 2 inverted tissue culture microscope
• Leica DMIL LED inverted fluorescence microscope with digital camera
• Nanodrop1
• T100 Thermal cycler
• Two biological safety cabinets
• Three cryobiological storage vessels
• Applied Biophysics Z-theta electric cell substrate impedance sensing (ECIS)
• Chromium X single cell RNA seq

Equipment available for use in the Molecular portion includes

• Spectromax Plus 364 spectrophotometer and plate reader for absorbance
• Spectromax Gemini EM plate reader for fluorescence
• Mastercycler PCR
• Bio-Rad CFX Connect Real-time PCR
• Applied Biosystems 7300 PCR
• FluorChem E imager
• Qubit 3.0 Fluorometer
• Nexcelom Cellometer Vision

Please complete the work order and submit to core manager.
 

Pricing information

Hood Usage: $5.00/hour
Liquid Nitrogen Storage: $5.00/month
ECIS: $10.00/day
Single cell RNA seq: $10.00/sample

Manoranjan Santra 
Module Manager
Department of Ophthalmology, Visual and Anatomical Sciences
7350 Scott Hall
Phone: 313-577-0748

Jena Steinle, Ph.D.
Module Director
Department of Ophthalmology, Visual and Anatomical Sciences
9314 Scott Hall
Phone: 313-577-9731