Retinal Ganglion Cell Isolation Kit ( #130-096-209 #130-096-464 #130-096-463 #130-096-462 )

Retinal Ganglion Cell Isolation Kit

Overview

These kits and tool boxes have been developed for the isolation of rat retinal ganglion cells (RGCs).

Details

Background information
Retinal ganglion cells (RGCs) are neurons of the central nervous system, which transmit information from the retina to the visual center of the brain. RGCs have been used to study neuron-glia interactions and the mechanisms of neurodegeneration.^1–4

Detailed separation procedure
The separation procedure involves the depletion of endothelial cells and microglia using MACSiBead™ Particles, and the positive selection of CD90.1⁺ RGCs using CD90.1 MicroBeads.⁵
First, the cell suspension is incubated with a biotin-conjugated antibody, which binds to endothelial cells and microglia, and with CD90.1 MicroBeads labeling RGCs. In the next step, endothelial cells and microglia are magnetically labeled with Anti-Biotin MACSiBead Particles and depleted by using a MACSiMAG™ Separator. Subsequently, the cell fraction depleted of endothelial cells and microglia is applied to MS Columns for positive selection of the CD90.1⁺ RGCs.

Note: MACSiBead Particles are not suitable for separations using MACS Columns and MiniMACS™, MidiMACS™, VarioMACS™, SuperMACS™, or autoMACS® Separators.

Columns

For positive selection: MS Columns

Further information

Dissection of retinas
[PDF; 71,8 KB]
MACS&more 12-2 (2010) Special edition neuroscience
[PDF; 4,7 MB]
Neuroscience research brochure
[PDF; 1,7 MB]

Figure 1

Isolation of CD90.1⁺ RGCs from postnatal day 7 rat retinal tissue using the Neural Tissue Dissociation Kit - Postnatal Neurons, the Retinal Ganglion Cell Isolation Kit, a MACSiMAG Separator, an OctoMACS™ Separator, and MS Columns. Cells were fluorescently stained with CD48-PE and CD90.1-FITC and analyzed by flow cytometry using the MACSQuant® Analyzer. Cell debris and dead cells were excluded from the analysis based on scatter signals and propidium iodide fluorescence.

A: Before separation

B: CD90.1⁺ retinal ganglion cells

CD90.1<sup>+</sup> retinal ganglion cells