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Flow Cytometry in Neuroscience Research

  • Charles KuszynskiEmail author
Protocol
  • 3.9k Downloads
Part of the Springer Protocols Handbooks book series (SPH)

Abstract

Flow cytometry has been used for numerous applications in immunology, cell biology, and neuroscience research (de Graaf, et al. 2011). With the development of higher sensitivity instrumentation and smaller numbers of cells required, flow cytometry has become an adjunctive methodology for studies of the interface between peripheral immune cells and the nervous system Schwatrz and Kipnis, 2011 (Ref). In this chapter, we will explore the use of flow cytometry to assess and isolate lymphoid cells associated with neuronal injury as well as how this approach would prove helpful in on the identification of neuronal cells. In addition, isolation of cells by other means will be discussed.

Keywords

Flow cytometry neuroscience Immunological staining 

References

  1. Badie B, Schartner JM, Paul J, Bartley BA, Jl V, Preston K (2000) Dexamethasone-induced abolition of the inflammatory response in an experimental glioma model: a flow cytometry study. J Neurosurg 93:634–639CrossRefPubMedGoogle Scholar
  2. Behbahani H, Rickle A, Concha H, Ankarcrona M, Winblad B, Cowburn RF (2005) Flow cytometry as a method for studying effects of stressors on primary rat neurons. J Neurosci Res 82:432–441CrossRefPubMedGoogle Scholar
  3. Bhattacharya S, Jackson JD, Das AV, Thoreson WB, Kuszynski C, James J, Joshi S, Ahmad I (2002) Direct identification and enrichment of retinal stem cells/progenitors by Hoechst 33342 dye efflux assay. Invest Opthalmol Vis Sci 44(6):2764–2773CrossRefGoogle Scholar
  4. Bilsland JG, Haldon C, Goddard J, Oliver K, Murray F, Wheeldon A, Cumberbatch J, McAllister G, Munoz-Sanjuan I (2006) A rapid method for the quantification of mouse hippocampal neurogenesis in vivo by flow cytometry: validation with conventional and enhanced immunohistochemical methods. J Neurosci Methods 157:54–63CrossRefPubMedGoogle Scholar
  5. de Graaf MT, de Jongste AHC, Kraan J, Boonstra JG, Sillevis Smitt PAE, Gratama JW (2011) Flow cytometric characterization of cerebrospinal fluid cells. Cytometry B Clin Cytom 80B:271–281CrossRefGoogle Scholar
  6. Gylys KH, Fein JA, Cole GM (2000) Quantitative characterization of crude synaptosomal fraction (P-2) components by flow cytometry. J Neurosci Res 61:186–192CrossRefPubMedGoogle Scholar
  7. Jamur MC, Oliver C (2010) Permeabilization of cell membranes. Methods Mol Biol 588:63–66CrossRefPubMedGoogle Scholar
  8. Jung SS, Nalbantoglu J, Cashman NR (1996) Alzheimer’s beta-amyloid precursor protein is expressed on the surface of immediately ex vivo brain cells: a flow cytometric study. J Neurosci Res 46:336–348CrossRefPubMedGoogle Scholar
  9. Kawaguchi A, Miyata T, Sawamoto K, Takashita N, Murayama A, Akamatsu W, Ogawa M, Okabe M, Tano Y, Goldman S, Okano H (2001) Nestin-EGFP transgenic mice: visualization of the self-renewal and multipotency of CNS stem cells. Mol Cell Neurosci 17:259–273CrossRefPubMedGoogle Scholar
  10. Kipnis J, Gadani S, Derecki NC (2012 Sep) Pro-cognitive properties of T cells. Nat Rev Immunol 12(9):663–669. doi: 10.1038/nri3280. Epub 2012 Aug 20, PubMed PMID: 22903149CrossRefPubMedPubMedCentralGoogle Scholar
  11. Klassen H, Schwartz MR, Bailey A, Young M (2001) Surface markers expressed by multipotent human and mouse neural progenitor cells include tetraspanins and non-protein epitopes. Neurosci Lett 312:180–182CrossRefPubMedGoogle Scholar
  12. Kraan J, Gratama JW, Haioun C, Orfao A, Plonquet A, Porwit A, Quijano S, Stetler-Stevenson M, Subira D, Wilson W (2008) Flow cytometric immunophenotyping of cerebrospinal fluid. Curr Protoc Cytom Chapter 6:Unit 6.25PubMedGoogle Scholar
  13. McLarena FH, Svendsenb CN, Van der Meidec P, Jolya E (2001) Analysis of neural stem cells by flow cytometry: cellular differentiation modifies patterns of MHC expression. J Neuroimmunol 112:35–46CrossRefGoogle Scholar
  14. Mosley RL, Hutter-Saunders JA, Stone DK, Gendelman HE (2012) Inflammation and adaptive immunity in Parkinson's disease. Cold Spring Harb Perspect Med 2(1):a009381, PMID:22315722CrossRefPubMedPubMedCentralGoogle Scholar
  15. Neurauter AA et al (2007) Cell isolation and expansion using Dynabeads®. Adv Biochem Eng Biotechnol 106:41–73PubMedGoogle Scholar
  16. Schwartz M, Kipnis JA (2011) A conceptual revolution in the relationships between the brain and immunity. Brain Behav Immun 25(5):817–819, PubMed PMID: 21187141; PubMed Central PMCID: PMC3074045CrossRefPubMedGoogle Scholar
  17. Stevens SL, Bao J, Hollis J, Lessov NS, Clark WM, Stenzel-Poorea MP (2002) The use of flow cytometry to evaluate temporal changes in inflammatory cells following focal cerebral ischemia in mice. Brain Res 932:110–119CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  1. 1.Department of Pathology and MicrobiologyUniversity of Nebraska Medical CenterOmahaUSA
  2. 2.Cell Analysis FacilityUniversity of Nebraska Medical CenterOmahaUSA

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