PMID- 23662303
OWN - NLM
STAT- MEDLINE
DCOM- 20131119
LR  - 20151119
IS  - 1364-5528 (Electronic)
IS  - 0003-2654 (Linking)
VI  - 138
IP  - 14
DP  - 2013 Jul 21
TI  - Determination of cell cycle phases in live B16 melanoma cells using IRMS.
PG  - 4015-21
LID - 10.1039/c3an00318c [doi]
AB  - The knowledge of cell cycle phase distribution is of paramount importance for 
      understanding cellular behaviour under normal and stressed growth conditions. 
      This task is usually assessed using Flow Cytometry (FC) or immunohistochemistry. 
      Here we report on the use of FTIR microspectroscopy in Microfluidic Devices 
      (MD-IRMS) as an alternative technique for studying cell cycle distribution in 
      live cells. Asynchronous, S- and G0-synchronized B16 mouse melanoma cells were 
      studied by running parallel experiments based on MD-IRMS and FC using Propidium 
      Iodide (PI) staining. MD-IRMS experiments have been done using silicon-modified 
      BaF2 devices, where the thin silicon layer prevents BaF2 dissolution without 
      affecting the transparency of the material and therefore enabling a better 
      assessment of the Phosphate I (PhI) and II (PhII) bands. Hierarchical Cluster 
      Analysis (HCA) of cellular microspectra in the 1300-1000 cm(-1) region pointed 
      out a distribution of cells among clusters, which is in good agreement with FC 
      results among G0/G1, S and G2/M phases. The differentiation is mostly driven by 
      the intensity of PhI and PhII bands. In particular, PhI almost doubles from the 
      G0/G1 to G2/M phase, in agreement with the trend followed by nucleic acids during 
      cellular progression. MD-IRMS is then proposed as a powerful method for the in 
      situ determination of the cell cycle stage of an individual cell, without any 
      labelling or staining, which gives the advantage of possibly monitoring specific 
      cellular responses to several types of stimuli by clearly separating the spectral 
      signatures related to the cellular response from those of cells that are normally 
      progressing.
FAU - Bedolla, Diana E
AU  - Bedolla DE
AD  - Elettra Sincrotrone Trieste, SISSI beamline, S.S. 14 Km 163.5, 34149 Basovizza, 
      Trieste, Italy.
FAU - Kenig, Sasa
AU  - Kenig S
FAU - Mitri, Elisa
AU  - Mitri E
FAU - Ferraris, Paolo
AU  - Ferraris P
FAU - Marcello, Alessandro
AU  - Marcello A
FAU - Grenci, Gianluca
AU  - Grenci G
FAU - Vaccari, Lisa
AU  - Vaccari L
LA  - eng
PT  - Journal Article
DEP - 20130510
PL  - England
TA  - Analyst
JT  - The Analyst
JID - 0372652
RN  - 0 (Nucleic Acids)
RN  - 36015-30-2 (Propidium)
SB  - IM
MH  - Animals
MH  - Cell Cycle/*physiology
MH  - *Cell Physiological Phenomena
MH  - Cluster Analysis
MH  - Flow Cytometry
MH  - Melanoma, Experimental/genetics/*pathology
MH  - Mice
MH  - Microfluidic Analytical Techniques/*instrumentation
MH  - Nucleic Acids/*analysis
MH  - Propidium
MH  - Spectroscopy, Fourier Transform Infrared/*methods
MH  - Tumor Cells, Cultured
EDAT- 2013/05/11 06:00
MHDA- 2013/11/20 06:00
CRDT- 2013/05/11 06:00
PHST- 2013/05/11 06:00 [entrez]
PHST- 2013/05/11 06:00 [pubmed]
PHST- 2013/11/20 06:00 [medline]
AID - 10.1039/c3an00318c [doi]
PST - ppublish
SO  - Analyst. 2013 Jul 21;138(14):4015-21. doi: 10.1039/c3an00318c. Epub 2013 May 10.