The Joint Cell Isolation Core

 Objective

The goal of the Joint Cell Isolation (JCI) Core is to provide MARC members with services for the isolation of tissue fragments or single cells from human or animal joint specimens.
 
The JCI Core has two components: the laser capture microdissection (LCM) component and the cell sorting component. By leveraging both flow cytometry and LCM capabilities, researchers at the MARC JCI Core can target a wide spectrum of isolates, ranging from single T cells in mouse synovium to cartilage and tendon tissue from human joints. To achieve successful and reproducible results, the JCI Core staff will be available to assist members with every stage of their experiments, ranging from design consultation to direct experimental facilitation.
 
Laser Capture Microdissection
The LCM component of the MARC JCI Core provides its users with operator-assisted laser capture of tissues or single cells from sections of human or animal joints. LCM is a method by which specific tissues and even single cells can be isolated from complex multi-tissue structures. Samples are prepared as histologically- or immunofluorescently-stained slides and visualized under the LCM microscope. The region of interest is selected and excised using an infrared (IR) or ultraviolet (UV) laser. This process can be repeated on the same prepared sample for the separate isolation of different tissue substructures or regions.
 
LCM Equipment
Applied Biosystems ArcturusXT laser capture microdissection system
-Nikon Ti-E microscope for imaging
-UV and IR solid-state lasers for sample excision
 
Services
Initial consultation up to 1 hr:  free of charge
Consultation and experimental design
Operator-assisted microdissection for established protocols
Collaborative operator-assisted establishment of new protocols (discounted rate)
Closing report upon completion of experimental series
 
Cell Sorting
The cell sorting component of the MARC JCI Core serves as a versatile state-of-the-art resource for sorting of subpopulations from human or animal joints. Integrated with the La Jolla Institute (LJI) Flow Sorting Facility, the Core has four cell sorters, each capable of separating populations into a variety of tubes or multi-well plates. These sorters will be made available to MARC members during normal hours, with the option of short-notice after-hours sorting appointments (prioritized for experiments with human clinical samples).
 
Sorters
FACSAria-1,2
-Violet (405 nm), blue (488 nm) and red (633 nm) lasers
-Capable of detecting 9 fluorescent parameters and 2 light scatter parameters
 
FACSAria-3
-Violet (405 nm), blue (488 nm), green/yellow (561 nm), and red (633 nm) lasers
-Capable of detecting 13 fluorescent parameters and 2 light scatter parameters
 
FACSAria-Fusion
-Violet (405 nm), blue (488 nm), green/yellow (561 nm), and red (633 nm) solid-state lasers
-Capable of detecting 16 fluorescent parameters and 2 light scatter parameters
-Safety measures for BSL-2+ and BSL-3 samples
 
Services
MARC members will receive the LJI rate for internal users, and the LJI Flow Core has set aside short notice after-hours sort time for our users. The Core can also perform joint disaggregation services to better facilitate sorting individual cells from joints for analysis.
 
Initial consultation:  free of charge
Consultation and experimental design
Joint tissue disaggregation
Collaborative operator-assisted establishment of new tissue disaggregation protocols (discounted rate)
Staff-operated cell sorting
User-operated cell sorting
Closing report upon completion of experimental series
 
Core Leadership & Contacts
Nunzio Bottini, MD, PhD, Core Co-Director
Stephanie Stanford, PhD, Core Co-Director
 
Benjamin Van Espen, JCI Core Manager
bvanespen@ucsd.edu
 
Alex Funderburk, MARC Coordinator
marcadmin@ucsd.edu
 
Service Request
To request service, please e-mail the Core Manager with the details of your experiment.
 
Other Resource Cores
·      Joint Bioinformatics and Computational Core
·      Joint Imaging Core