Combining combing and secondary ion mass spectrometry to study DNA on chips using (13)C and (15)N labeling.

Dynamic secondary ion mass spectrometry ( D-SIMS) imaging of combed DNA - the combing, imaging by SIMS or CIS method - has been developed previously using a standard NanoSIMS 50 to reveal, on the 50 nm scale, individual DNA fibers labeled with different, non-radioactive isotopes in vivo and to quantify these isotopes. This makes CIS especially suitable for determining the times, places and rates of DNA synthesis as well as the detection of the fine-scale re-arrangements of DNA and of molecules associated with combed DNA fibers. Here, we show how CIS may be extended to (13)C-labeling via the detection and quantification of the (13)C (14)N (-) recombinant ion and the use of the (13)C: (12)C ratio, we discuss how CIS might permit three successive labels, and we suggest ideas that might be explored using CIS.

Role of parietal and principal gastric mucosa cells in the phenomenon of concentration of aluminum and indium.

The subcellular behavior of aluminum and indium, used in medical and industrial fields, was studied in the gastric mucosa and the liver after their intragastric administration to rats, using, two of the most sensitive methods of observation and microanalysis, the transmission electron microscopy, and the secondary ion mass spectrometry. The ultrastructural study showed the presence of electron dense deposits, in the lysosomes of parietal and principal gastric mucosa cells but no loaded lysosomes were observed in the different studied hepatic territories. The microanalytical study allowed the identification of the chemical species present in those deposits as aluminum or indium isotopes and the cartography of their distribution. No modification was observed in control rats tissues. In comparison to previous studies describing the mechanism of aluminum concentration in the gastric mucosa and showing that this element was concentrated in the lysosomes of fundic and antral human gastric mucosa, our study provided additional informations about the types of cells involved in the phenomenon of concentration of aluminum and indium, which are the parietal and the principal cells of the gastric mucosa. Our study demonstrated that these cells have the ability to concentrate selectively aluminum and indium in their lysosomes, as a defensive reaction against intoxication by foreign elements.

Combed single DNA molecules imaged by secondary ion mass spectrometry.

Studies of replication, recombination, and rearrangements at the level of individual molecules of DNA are often limited by problems of resolution or of perturbations caused by the modifications that are needed for imaging. The Combing-Imaging by Secondary Ion Mass Spectrometry (SIMS) (CIS) method helps solve these problems by combining DNA combing, cesium flooding, and quantitative imaging via the NanoSIMS 50. We show here that CIS can reveal, on the 50 nm scale, individual DNA fibers labeled with different, nonradioactive isotopes and, moreover, that it can quantify these isotopes so as to detect and measure the length of one or more short nucleic acid fragments associated with a longer fiber.

Dynamic NanoSIMS ion imaging of unicellular freshwater algae exposed to copper.

This work demonstrates the capabilities of nanoscale secondary-ion mass spectrometry, using the Cameca NanoSIMS50 ion microprobe, to detect and image the copper-ion distribution in microalgal cells exposed to nanomolar and micromolar copper concentrations. In parallel to (63)Cu(-) secondary-ion maps, images of (12)C(-), (12)C(14)N(-), and (31)P(-) secondary ions were collected and analysed. A correlation of (63)Cu(-) secondary-ion maps with those found for (12)C(14)N(-) and (31)P(-) demonstrated the possible association of Cu with cell components rich in proteins and phosphorus. The results highlighted the potential of NanoSIMS for intracellular tracking of essential trace elements such as Cu in single cells of the microalga Chlorella kesslerii.