PMID- 25420714
OWN - NLM
STAT- MEDLINE
DCOM- 20151123
LR  - 20191210
IS  - 1618-2650 (Electronic)
IS  - 1618-2642 (Linking)
VI  - 407
IP  - 8
DP  - 2015 Mar
TI  - Characterization of freeze-fractured epithelial plasma membranes on nanometer 
      scale with ToF-SIMS.
PG  - 2203-11
LID - 10.1007/s00216-014-8334-2 [doi]
AB  - Time-of-flight secondary ion mass spectrometry (ToF-SIMS) was used to 
      characterize the freeze-fracturing process of human epithelial PANC-1 and UROtsa 
      cells. For this purpose, phosphatidylcholine, sphingomyelin, 
      phosphatidylethanolamine, and phosphatidylserine standard samples were 
      investigated to find specific signals with both high specificity and signal 
      intensity. The results were used to investigate single cells of subconfluent cell 
      layers prepared with a special silicon wafer sandwich preparation technique. This 
      freeze-fracturing technique strips cell membranes off the cells, isolating them 
      on opposing silicon wafer substrates. Criteria were found for defining regions 
      with stripped off cell membranes and, on the opposing wafer, complementary 
      regions with the remaining cells. Measured ethanolamine/choline and 
      serine/choline ratios in these regions clearly showed that in the 
      freeze-fracturing process, the lipid bilayer of the plasma membrane is split 
      along its central zone. Accordingly, only the outer lipid monolayer is stripped 
      off the cell, while the inner lipid monolayer remains attached to the cell on the 
      opposing wafer, thus allowing detailed analysis of a single lipid monolayer. 
      Furthermore, it could be shown that using different washing procedures did not 
      influence the transmembrane lipid distribution. Under optimized preparation 
      conditions, it became feasible to detect lipids with a lateral resolution of 
      approximately 100 nm. The data indicate that ToF-SIMS would be a very useful 
      technique to study with very high lateral resolution changes in lipid composition 
      caused, for example, by lipid storage diseases or pharmaceuticals that interfere 
      with the lipid metabolism.
FAU - Draude, Felix
AU  - Draude F
AD  - Physikalisches Institut, Westfalische Wilhelms-Universitat Munster, 
      Wilhelm-Klemm-Str. 10, 48149, Munster, Germany.
FAU - Korsgen, Martin
AU  - Korsgen M
FAU - Pelster, Andreas
AU  - Pelster A
FAU - Schwerdtle, Tanja
AU  - Schwerdtle T
FAU - Muthing, Johannes
AU  - Muthing J
FAU - Arlinghaus, Heinrich F
AU  - Arlinghaus HF
LA  - eng
PT  - Evaluation Study
PT  - Journal Article
DEP - 20141125
PL  - Germany
TA  - Anal Bioanal Chem
JT  - Analytical and bioanalytical chemistry
JID - 101134327
RN  - 0 (Lipid Bilayers)
RN  - 0 (Lipids)
SB  - IM
MH  - Cell Line, Tumor
MH  - Cell Membrane/*chemistry/metabolism
MH  - Epithelial Cells/*chemistry/metabolism
MH  - Freeze Fracturing
MH  - Humans
MH  - Lipid Bilayers/chemistry
MH  - Lipid Metabolism
MH  - Lipids/chemistry
MH  - Spectrometry, Mass, Secondary Ion/*methods
EDAT- 2014/11/26 06:00
MHDA- 2015/12/15 06:00
CRDT- 2014/11/26 06:00
PHST- 2014/09/12 00:00 [received]
PHST- 2014/11/07 00:00 [accepted]
PHST- 2014/11/05 00:00 [revised]
PHST- 2014/11/26 06:00 [entrez]
PHST- 2014/11/26 06:00 [pubmed]
PHST- 2015/12/15 06:00 [medline]
AID - 10.1007/s00216-014-8334-2 [doi]
PST - ppublish
SO  - Anal Bioanal Chem. 2015 Mar;407(8):2203-11. doi: 10.1007/s00216-014-8334-2. Epub 
      2014 Nov 25.