PMID- 17119992
OWN - NLM
STAT- MEDLINE
DCOM- 20080108
LR  - 20181113
IS  - 0009-5915 (Print)
IS  - 1432-0886 (Electronic)
IS  - 0009-5915 (Linking)
VI  - 116
IP  - 2
DP  - 2007 Apr
TI  - Preservation of large-scale chromatin structure in FISH experiments.
PG  - 117-33
AB  - The nuclear organization of specific endogenous chromatin regions can be 
      investigated only by fluorescence in situ hybridization (FISH). One of the two 
      fixation procedures is typically applied: (1) buffered formaldehyde or (2) 
      hypotonic shock with methanol acetic acid fixation followed by dropping of nuclei 
      on glass slides and air drying. In this study, we compared the effects of these 
      two procedures and some variations on nuclear morphology and on FISH signals. We 
      analyzed mouse erythroleukemia and mouse embryonic stem cells because their 
      clusters of subcentromeric heterochromatin provide an easy means to assess 
      preservation of chromatin. Qualitative and quantitative analyses revealed that 
      formaldehyde fixation provided good preservation of large-scale chromatin 
      structures, while classical methanol acetic acid fixation after hypotonic 
      treatment severely impaired nuclear shape and led to disruption of chromosome 
      territories, heterochromatin structures, and large transgene arrays. Our data 
      show that such preparations do not faithfully reflect in vivo nuclear 
      architecture.
FAU - Hepperger, Claudia
AU  - Hepperger C
AD  - Department Biologie II, Ludwig-Maximilians-Universitat Munchen, Grosshaderner 
      Str. 2, 82152 Planegg-Martinsried, Germany.
FAU - Otten, Simone
AU  - Otten S
FAU - von Hase, Johann
AU  - von Hase J
FAU - Dietzel, Steffen
AU  - Dietzel S
LA  - eng
PT  - Comparative Study
PT  - Journal Article
PT  - Research Support, Non-U.S. Gov't
DEP - 20061122
PL  - Austria
TA  - Chromosoma
JT  - Chromosoma
JID - 2985138R
RN  - 0 (Chromatin)
RN  - 1HG84L3525 (Formaldehyde)
SB  - IM
MH  - Animals
MH  - Cell Line, Tumor
MH  - Cell Nucleus/*ultrastructure
MH  - Chromatin/*chemistry
MH  - Embryonic Stem Cells
MH  - Formaldehyde
MH  - Image Processing, Computer-Assisted
MH  - In Situ Hybridization, Fluorescence/*methods
MH  - Mice
MH  - Microscopy, Confocal
MH  - Tissue Fixation/*methods
PMC - PMC1824788
EDAT- 2006/11/23 09:00
MHDA- 2008/01/09 09:00
PMCR- 2007/03/15
CRDT- 2006/11/23 09:00
PHST- 2006/05/30 00:00 [received]
PHST- 2006/09/23 00:00 [accepted]
PHST- 2006/08/11 00:00 [revised]
PHST- 2006/11/23 09:00 [pubmed]
PHST- 2008/01/09 09:00 [medline]
PHST- 2006/11/23 09:00 [entrez]
PHST- 2007/03/15 00:00 [pmc-release]
AID - 84 [pii]
AID - 10.1007/s00412-006-0084-2 [doi]
PST - ppublish
SO  - Chromosoma. 2007 Apr;116(2):117-33. doi: 10.1007/s00412-006-0084-2. Epub 2006 Nov 
      22.