PMID- 35218531
OWN - NLM
STAT- MEDLINE
DCOM- 20220301
LR  - 20230217
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 2440
DP  - 2022
TI  - Three-Dimensional Simultaneous Imaging of Nucleic Acids and Proteins During 
      Influenza Virus Infection in Single Cells Using Confocal Microscopy.
PG  - 41-56
LID - 10.1007/978-1-0716-2051-9_2 [doi]
AB  - Three-dimensional imaging is a powerful tool for examining the spatial 
      distribution of intracellular molecules like nucleic acids, proteins, and 
      organelles in cells and tissues. Multicolor fluorescence imaging coupled with 
      three-dimensional spatial information provide a platform to explore the 
      relationship between different cellular features and molecules. We have 
      previously developed a pipeline to study the intracellular localization of 
      influenza virus genomic segments within an infected cell. Here, we describe the 
      staining of multiple viral RNA segments in cells infected with influenza virus by 
      combined fluorescence in situ hybridization (FISH) and immunofluorescence and 
      quantification of colocalization between viral segments. This chapter will cover 
      the acquisition and analysis of 3D images by the widely used laser scanning 
      confocal microscope. These strategies can be applied to a wide range of 
      biological processes and modified to examine colocalization of other cellular 
      features.
CI  - (c) 2022. The Author(s), under exclusive license to Springer Science+Business 
      Media, LLC, part of Springer Nature.
FAU - Manivanh, Richard
AU  - Manivanh R
AD  - Department of Microbiology & Molecular Genetics, University of Pittsburgh School 
      of Medicine, Pittsburgh, PA, USA.
FAU - Lakdawala, Seema S
AU  - Lakdawala SS
AD  - Department of Microbiology & Molecular Genetics, University of Pittsburgh School 
      of Medicine, Pittsburgh, PA, USA.
FAU - Jones, Jennifer E
AU  - Jones JE
AD  - Department of Microbiology & Molecular Genetics, University of Pittsburgh School 
      of Medicine, Pittsburgh, PA, USA. jenny@jennyjonesphd.com.
LA  - eng
PT  - Journal Article
PT  - Research Support, N.I.H., Extramural
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Nucleic Acids)
SB  - IM
MH  - Humans
MH  - Imaging, Three-Dimensional/methods
MH  - In Situ Hybridization, Fluorescence/methods
MH  - *Influenza, Human
MH  - Microscopy, Confocal/methods
MH  - *Nucleic Acids
MH  - *Orthomyxoviridae
OTO - NOTNLM
OT  - 3D imaging
OT  - Confocal microscopy
OT  - Deconvolution
OT  - Fluorescence in situ hybridization
OT  - Fluorescence microscopy
OT  - Huygens
OT  - Image acquisition
OT  - Imaris
OT  - Influenza virus
OT  - Multicolor
OT  - Point spread function
EDAT- 2022/02/27 06:00
MHDA- 2022/03/03 06:00
CRDT- 2022/02/26 12:07
PHST- 2022/02/26 12:07 [entrez]
PHST- 2022/02/27 06:00 [pubmed]
PHST- 2022/03/03 06:00 [medline]
AID - 10.1007/978-1-0716-2051-9_2 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2022;2440:41-56. doi: 10.1007/978-1-0716-2051-9_2.