NanoSIMS 50 in biological field. - CAMECA - #2

SecondarylonMassSpectrometryisbasedupon the sputtering ofa few atomic layers from the surface of a sample, induced by a 'primary ion' bombardment. A primary ion impact triggers a cascade of atomic collisions. Atoms and atomic clusters are ejected. During the ejection process, some of them are spontaneously ionized. These 'secondary ions' are a characteristic of the c omposition of the analyzed area. They are separated ac c ording to their mass, and an image containing quantitative infomiation is formed fora selected mass.

The NanoSZMS 50 provides the capability of recording simultaneously several atomic mass images (parallel imaging) together with a high mass separation (M/dM>5000) at high transmission (70%) and a high spatial resolution (50nm). These characteristics allow comparison inexactly the same subc ellularc ompartments, of images ofthe distribution and measure, ment of the acc umulation of several eh me nts, or molec uks labeled with differe nt stable isotopes (e.g. ¹⁵N, ¹³C, ²H, ¹⁸0, ³³S, ⁷⁴Se, ...). Multiple labeling can be used on different or on the same molecules.

Multiple Isotope Mass Spectrometry (MIMS) offers a novel, powerful and quantitative technique forstudying intra-and trans-c ellularmetabolic pathways, signal transduction, cytoplasmic and nucleo-cytoplasmic translocations, UNA and DNA expression and distribution, vesicular and non-vesicular trafficking, vims penetration, and localization of drugs. MIMS opens a world of labeling possibMties that are impossible with autoradiography, and offers a much higher sensitivity. 50nm spatial resolution is by far superior to laser c onfoc al optical microscopy used with fiuoresc ent markers.

Chloroplasts from

c hlo ro p hyllia n cells in leaf of

Arab idop sis thaliana after pulsing

with 15-N. The use of 15-nitrogen

trac er allows to follow the

histologic al pathway of N

incorporationin the tissue s.

He Id: 10um x 10um.

Mouse c oc hlea: protein turnover study with 15-N marker. Simultaneous imaging of 14-N and 15-N. Nitro g e n iso to p e ratio wasmeasured from areas indicated onleftimage. Reissnermembrane (Rm), nucleus (N) and fourred blood ceUs(RBC). Held: 12um x 12um. Pr. C.Iechene and Pr. EMroz, Ha rva rd Me die al School*.

12_C14_N 32g 16Q

Ultra -struc tural c e 11 distrib utio n o f the melanoma marker iodobenzamide in mouse lung tissues, confirming the specific affinity of this molecule formelanin. The molecule is imaged through its Iodine atoms. Upperfield: 20 x 20um, ls>wer. 10 x 10um. J-L Guerq uin-Ke m, J.-C. Made Imo nt a nd Al, Bb Medical Engineering OnUne 2004, 3:10

Ißwerleft: Haircryo-section SEM image. Upper left and right: false colourimage of the distribution of C, O and Non melanine granules in the hairsection. Stable tracers (2D, 13C, 15N...) in growing medium or shampoo allow to monitorthe incorporation o f m o le c ule s in the ha ir.

Overlay of ²⁶CN (red), ³²S (green), ¹⁶0" (blue). Held: 3.5um x3.5um.