PMID- 20809314
OWN - NLM
STAT- MEDLINE
DCOM- 20101214
LR  - 20211028
IS  - 1940-6029 (Electronic)
IS  - 1064-3745 (Linking)
VI  - 659
DP  - 2010
TI  - CryoFISH: fluorescence in situ hybridization on ultrathin cryosections.
PG  - 219-30
LID - 10.1007/978-1-60761-789-1_15 [doi]
AB  - The visualization of cellular structures and components has become an invaluable 
      tool in biological and medical sciences. Imaging subcellular compartments and 
      single molecules within a cell has prompted the development of a wide range of 
      sample preparation techniques as well as various microscope devices to obtain 
      images with increased spatial resolution. Here, we present cryoFISH, a method for 
      fluorescence in situ hybridization (FISH) on thin ( approximately 150 nm thick) 
      cryosections from sucrose-embedded fixed cells or tissues. CryoFISH can be used 
      in combination with immunodetection (IF) of other cellular components. The main 
      advantages of cryoFISH and cryoIF over whole-cell labeling methods are increased 
      spatial resolution with confocal microscopy, greater sensitivity of detection due 
      to increased probe accessibility, and better image contrast. CryoFISH and cryoIF 
      methods typically used on samples fixed in conditions that preserve 
      ultrastructure, are compatible with the labeling of cells in their tissue context 
      and are ideal for correlative studies that compare fluorescence with electron 
      microscopy.
FAU - Xie, Sheila Q
AU  - Xie SQ
AD  - MRC Clinical Sciences Centre, Imperial College School of Medicine, London, UK.
FAU - Lavitas, Liron-Mark
AU  - Lavitas LM
FAU - Pombo, Ana
AU  - Pombo A
LA  - eng
GR  - MC_U120061476/MRC_/Medical Research Council/United Kingdom
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
PL  - United States
TA  - Methods Mol Biol
JT  - Methods in molecular biology (Clifton, N.J.)
JID - 9214969
RN  - 0 (Antigens)
RN  - 0 (Immunoglobulin M)
RN  - EC 2.7.7.- (RNA Polymerase II)
SB  - IM
MH  - Alleles
MH  - Animals
MH  - Antigens/immunology
MH  - Base Sequence
MH  - Cell Nucleus/metabolism
MH  - Chromosomes/genetics/metabolism
MH  - Cryoultramicrotomy/*methods
MH  - Humans
MH  - Immunoassay
MH  - Immunoglobulin M/immunology
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Mice
MH  - Phosphorylation
MH  - RNA Polymerase II/chemistry/metabolism
MH  - Tissue Embedding
MH  - Tissue Fixation
EDAT- 2010/09/03 06:00
MHDA- 2010/12/16 06:00
CRDT- 2010/09/03 06:00
PHST- 2010/09/03 06:00 [entrez]
PHST- 2010/09/03 06:00 [pubmed]
PHST- 2010/12/16 06:00 [medline]
AID - 10.1007/978-1-60761-789-1_15 [doi]
PST - ppublish
SO  - Methods Mol Biol. 2010;659:219-30. doi: 10.1007/978-1-60761-789-1_15.