Using Flow Cytometry in Neuroscience Research


Sample Preparation

One of the initial hurdles in establishing flow cytometry applications in neuroscience was preparing suitable samples. When using whole brain tissue, generation of single cell suspensions is possible through a variety of methods.

 

 

 
Mechanical Dissociation

First, tissues need to be mechanically dissociated into small fragments using a razor blade prior to processing.

 
Enzymatic Digestion

Enzymes such as Trypsin, Accutase®, or Papain can be used to digest whole brain tissue samples.
Density Gradient

Percoll® or sucrose gradients help to remove myelin from the sample. For this, we use a 70/37/30% Percoll® gradient.

 
Cell Markers

Flow cytometry in neuroscience is useful when you need to study or isolate a homogenous population from a heterogeneous sample, such as primary cell suspensions or in vitro differentiation of neural stem cell cultures. Identification of cell type-specific markers allows for detection or isolation of these cells through magnetic cell separation or cell sorting. Cell type-specific markers have been identified for a variety of neural cell types including astrocytes, microglia, and oligodendrocytes.

 

 

 
CD56 (NCAM)
CD24
CD200
CD11b
CD45
CX3CR1
P2RY12
TMEM119
GFAP
GLAST

 

 
A2B5
PDGFRα
NG2
CD15
CD29
CD133
CD184
CD171
Nestin
Notch-1
Notch-2
Sox-2

Check out these references using flow cytometry to identify neural cell types:


 

Organelle Markers & Function

Altered organelle morphology and health is involved in numerous neurodegenerative diseases. For example, mitochondrial dysfunction and accumulation of reactive oxygen species is common in Alzheimer's and Parkinson's disease. When performing flow cytometry, in conjunction with cell type-specific markers, you can include a cell-permeant dye like MitoSpy™ as an indicator of cell health and mitochondrial function.

Similarly, lysosomal proteins help clear aggregated proteins associated with disease. Antibodies against lysosome markers like LAMP-1 or LAMP-2 can be used in flow cytometry to compare lysosome function in normal and disease states.

Find your antibodies for Neuroscience and Flow Cytometry at: biolegend.com/neuroscience
Contributed by Kelsey Swartz, PhD.
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