Development of the CAS-LIBB single-particle microbeam for localized irradiation of living cells

Abstract

A single-particle microbeam facility has been constructed at the Key Laboratory of Ion Beam Bioengineering (LIBB), Chinese Academy of Sciences (CAS). The system was designed to deliver a defined numbers of hydrogen ions, produced by a van de Graaff accelerator, in an energy range of 2.0-3.0 MeV, into an area smaller than that of the nucleus of an individual living cell. The beam is collimated by a borosilicate glass capillary that forms the beam-line exit. An integrated computer program recognizes the cells and locates them one by one over the microbeam exit for irradiation. We present technical details of the CAS-LIBB microbeam facility, particularly on the collimator, hardware, control program, as well as cell irradiation protocols available. Various factors contributing to the targeting and positioning precision are discussed along with accuracy measurement results.

Akt/mTOR mediated induction of bystander effect signaling in a nucleus independent manner in irradiated human lung adenocarcinoma epithelial cells

“…A549 cells were denucleated according to the methods described in ref. [ 15 ], and then the A549 cytoplasts were irradiated precisely with the microbeam facility at CAS-LIBB, which allowed individual protons to be delivered to cells with high reproducibility (single ion delivered with 99% efficiency) and high accuracy (99% within 5 μm) [ 16 ]. About 1,000 fluorescent A549 cells/cytoplasts were seeded in the central area (5 mm in diameter) of a specially designed microbeam dish consisting of a 3.5 μm-thick polypropylene film base (Collaborative Biomedical Products, Bedford, MA, USA).…”

“…The microbeam facility at CAS-LIBB was used for this study [ 16 ]. The irradiated cells/cytoplasts were stained with Hoechst 33342 (10 μg/mL; Invitrogen, Eugene, OR, USA) and CellTracker orange CMRA (5 μmol/L; Life Technologies, Eugene, OR, USA), and the bystander cells were not stained.…”

Spatial function of the oxidative DNA damage response in radiation induced bystander effects in intra- and inter-system ofCaenorhabditis elegans

“…The proton microbeam facility in our laboratory (CASLIBB) delivering defined numbers of charged particles was used for the localized irradiation of C. elegans . The average energy range of incident protons was 2.0~3.0 MeV with a LET of 11 keV/μm and the average beam diameter on samples measured less than 10 μm using CR-39 solid detectors for 10,000 protons [ 38 ]. For microbeam irradiation, synchronized worms at the L4 stage were picked out and placed on 2% of agarose gels and then anesthetized as described previously [ 13 ].…”

Improved detection resolution in single particle microbeam system

“…Two 5 Â 5 grids of a pit cluster by 100-hit on the CR39 surface provide statistical assessment of the targeting accuracy of the different designs. The experimental details can be found elsewhere [29]. 6.25 M NaOH solution at 70 1C for 6 h. Fig.…”

“…As a result, there is a high demand for high energy resolution and spatial resolution. A typical particle detection system is composed of a scintillator and Al foil acting as a reflecting film in conjunction with a photomultiplier (PM) tube [21,[29][30][31]. However, when Al foils are used, energy loss and beam scattering become significant and the energy resolution and spatial resolution of the single particle beam will degrade.…”