Subcellular mass determination by 4He+ energy-loss micro-spectrometry.
The scanning transmission ion microscope (STIM) has been used to determine the intracellular mass of human cultured cells. A 4He+ microbeam of 2.0 MeV energy was chosen to obtain enhanced ion-energy-loss sensitivity through the micron-thick freeze-dried cells. Local sample mass calculation, based on energy-loss conversion by use of appropriate matrix stopping powers, was performed by use of dedicated software. The method was validated with epoxy resin sections and polymer foil as analogues of biological samples in the range of (intra)cellular thickness, 150 to 3000 nm. STIM analysis resulted in less than 5% error in mass determination. 4He+ energy-loss micro-spectrometry was performed on freeze-dried human ovarian cancer cells, the mean areal mass obtained was 120 microg cm(-2) (200 microg cm(-2) in the nucleus and 250 microg cm(-2) in nucleoli). This method is particularly useful for mass normalization of X-ray fluorescence yields resulting from particle-induced X-ray emission microanalysis (micro-PIXE). When performed successively these two ion-beam micro-analytical methods enable the mapping of true element concentrations within single cells.
Devès
Guillaume
G
Chimie Nucléaire Analytrique et Bioenvironnementale CNRS UMRS084, Université de Bordeaux 1, BP 120 Le Haut-Vigneau, 33175 Gradignan cedex, France.
Ortega
Richard
R
D000230:Adenocarcinoma / Q000737:chemistry*; D004853:Epoxy Resins / Q000032:analysis; D005260:Female; D006371:Helium / Q000737:chemistry; D006801:Humans; D017348:Microscopy, Electron, Scanning Transmission / Q000379:methods; D010051:Ovarian Neoplasms / Q000737:chemistry*; D011093:Polyethylene Terephthalates / Q000032:analysis; D013052:Spectrometry, X-Ray Emission / Q000379:methods*; D013054:Spectrophotometry, Atomic; D013347:Subcellular Fractions / Q000737:chemistry*
D004853:Epoxy Resins; D011093:Polyethylene Terephthalates; D006371:Helium