PMID- 28448013
OWN - NLM
STAT- MEDLINE
DCOM- 20180125
LR  - 20210109
IS  - 1940-087X (Electronic)
IS  - 1940-087X (Linking)
IP  - 122
DP  - 2017 Apr 5
TI  - Precision-cut Mouse Lung Slices to Visualize Live Pulmonary Dendritic Cells.
LID - 10.3791/55465 [doi]
LID - 55465
AB  - Inhalation of allergens and pathogens elicits multiple changes in a variety of 
      immune cell types in the lung. Flow cytometry is a powerful technique for 
      quantitative analysis of cell surface proteins on immune cells, but it provides 
      no information on the localization and migration patterns of these cells within 
      the lung. Similarly, chemotaxis assays can be performed to study the potential of 
      cells to respond to chemotactic factors in vitro, but these assays do not 
      reproduce the complex environment of the intact lung. In contrast to these 
      aforementioned techniques, the location of individual cell types within the lung 
      can be readily visualized by generating Precision-cut Lung Slices (PCLS), 
      staining them with commercially available, fluorescently tagged antibodies, and 
      visualizing the sections by confocal microscopy. PCLS can be used for both live 
      and fixed lung tissue, and the slices can encompass areas as large as a cross 
      section of an entire lobe. We have used this protocol to successfully visualize 
      the location of a wide variety of cell types in the lung, including distinct 
      types of dendritic cells, macrophages, neutrophils, T cells and B cells, as well 
      as structural cells such as lymphatic, endothelial, and epithelial cells. The 
      ability to visualize cellular interactions, such as those between dendritic cells 
      and T cells, in live, three-dimensional lung tissue, can reveal how cells move 
      within the lung and interact with one another at steady state and during 
      inflammation. Thus, when used in combination with other procedures, such as flow 
      cytometry and quantitative PCR, PCLS can contribute to a comprehensive 
      understanding of cellular events that underlie allergic and inflammatory diseases 
      of the lung.
FAU - Lyons-Cohen, Miranda R
AU  - Lyons-Cohen MR
AD  - Immunity, Inflammation, and Disease Laboratory, Division of Intramural Research, 
      National Institute of Environmental Health Sciences, NIH.
FAU - Thomas, Seddon Y
AU  - Thomas SY
AD  - Immunity, Inflammation, and Disease Laboratory, Division of Intramural Research, 
      National Institute of Environmental Health Sciences, NIH.
FAU - Cook, Donald N
AU  - Cook DN
AD  - Immunity, Inflammation, and Disease Laboratory, Division of Intramural Research, 
      National Institute of Environmental Health Sciences, NIH.
FAU - Nakano, Hideki
AU  - Nakano H
AD  - Immunity, Inflammation, and Disease Laboratory, Division of Intramural Research, 
      National Institute of Environmental Health Sciences, NIH; NakanoH@niehs.nih.gov.
LA  - eng
GR  - ZIA ES102025/ImNIH/Intramural NIH HHS/United States
GR  - ZIA ES102025-09/ImNIH/Intramural NIH HHS/United States
PT  - Journal Article
PT  - Video-Audio Media
DEP - 20170405
PL  - United States
TA  - J Vis Exp
JT  - Journal of visualized experiments : JoVE
JID - 101313252
RN  - 0 (Allergens)
SB  - IM
MH  - Allergens
MH  - Animals
MH  - Dendritic Cells/*immunology
MH  - Epithelial Cells/*immunology
MH  - Flow Cytometry
MH  - Fluorescent Antibody Technique
MH  - Histological Techniques/*methods
MH  - Lung/*cytology
MH  - Macrophages/immunology
MH  - Mice
MH  - Specimen Handling/methods
MH  - *Tissue Culture Techniques
PMC - PMC5512285
MID - NIHMS875636
EDAT- 2017/04/28 06:00
MHDA- 2018/01/26 06:00
PMCR- 2018/04/05
CRDT- 2017/04/28 06:00
PHST- 2017/04/28 06:00 [entrez]
PHST- 2017/04/28 06:00 [pubmed]
PHST- 2018/01/26 06:00 [medline]
PHST- 2018/04/05 00:00 [pmc-release]
AID - 55465 [pii]
AID - 10.3791/55465 [doi]
PST - epublish
SO  - J Vis Exp. 2017 Apr 5;(122):55465. doi: 10.3791/55465.