PMID- 29238736
OWN - NLM
STAT- PubMed-not-MEDLINE
LR  - 20220210
IS  - 2331-8325 (Print)
IS  - 2331-8325 (Electronic)
IS  - 2331-8325 (Linking)
VI  - 7
IP  - 22
DP  - 2017 Nov 20
TI  - Combination of Fluorescent in situ Hybridization (FISH) and Immunofluorescence 
      Imaging for Detection of Cytokine Expression in Microglia/Macrophage Cells.
LID - e2608 [pii]
LID - 10.21769/BioProtoc.2608 [doi]
AB  - Microglia and macrophage cells are the primary producers of cytokines in response 
      to neuroinflammatory processes. But these cytokines are also produced by other 
      glial cells, endothelial cells, and neurons. It is essential to identify the 
      cells that produce these cytokines to target their different levels of 
      activation. We used dual RNAscope((R)) fluorescence in situ hybridization (FISH) 
      and immunohistochemistry (IHC) techniques to visualize the mRNA expression 
      pattern of pro- and anti-inflammatory cytokines in microglia/macrophages cells. 
      Using these methods, we can associate one mRNA to specific cell types when 
      combining with different cellular markers by immunofluorescence. Results from 
      RNAscope((R)) probes IL-1beta, TNFalpha, TGFbeta, IL-10 or Arg1, showed colocalization with 
      antibodies for microglia/macrophage cells. These target probes showed adequate 
      sensitivity and specificity to detect mRNA expression. New FISH detection 
      techniques combined with immunohistochemical techniques will help to jointly 
      determine the protein and mRNA localization, as well as provide reliable 
      quantification of the mRNA expression levels.
FAU - Fe Lanfranco, Maria
AU  - Fe Lanfranco M
AD  - Department of Neuroscience, Georgetown University, Washington, DC, United States.
FAU - Loane, David J
AU  - Loane DJ
AD  - Department of Anesthesiology, Center for Shock, Trauma, and Anesthesiology 
      Research (STAR), University of Maryland School of Medicine, Baltimore, MD, United 
      States.
FAU - Mocchetti, Italo
AU  - Mocchetti I
AD  - Department of Neuroscience, Georgetown University, Washington, DC, United States.
FAU - Burns, Mark P
AU  - Burns MP
AD  - Department of Neuroscience, Georgetown University, Washington, DC, United States.
FAU - Villapol, Sonia
AU  - Villapol S
AD  - Department of Neuroscience, Georgetown University, Washington, DC, United States.
LA  - eng
GR  - R03 NS067417/NS/NINDS NIH HHS/United States
GR  - R03 NS095038/NS/NINDS NIH HHS/United States
GR  - R01 NS081068/NS/NINDS NIH HHS/United States
GR  - R01 NS082308/NS/NINDS NIH HHS/United States
GR  - R21 NS102121/NS/NINDS NIH HHS/United States
PT  - Journal Article
PL  - United States
TA  - Bio Protoc
JT  - Bio-protocol
JID - 101635102
PMC - PMC5725199
MID - NIHMS922226
OTO - NOTNLM
OT  - Cytokines
OT  - Immunofluorescence
OT  - Macrophages
OT  - Microglia cells
OT  - Neuroinflammation
OT  - RNAscope
OT  - Traumatic brain injury (TBI)
OT  - in situ hybridization
COIS- The authors declare that the research was conducted in the absence of any 
      commercial or financial relationships that could be construed as a potential 
      conflict of interest.
EDAT- 2017/12/15 06:00
MHDA- 2017/12/15 06:01
PMCR- 2018/11/20
CRDT- 2017/12/15 06:00
PHST- 2017/12/15 06:00 [entrez]
PHST- 2017/12/15 06:00 [pubmed]
PHST- 2017/12/15 06:01 [medline]
PHST- 2018/11/20 00:00 [pmc-release]
AID - e2608 [pii]
AID - 2608 [pii]
AID - 10.21769/BioProtoc.2608 [doi]
PST - ppublish
SO  - Bio Protoc. 2017 Nov 20;7(22):e2608. doi: 10.21769/BioProtoc.2608.