The INFN-LNL Istituto Nazionale di Fisica Nucleare-Laboratori Nazionali di Legnaro (partner #4) contribution to the project Roberto CHERUBINI, INFN-LNL CELLION Mid-term Meeting CENBG, Bordeaux,Feb 3-5, 2006 Silvia Gerardi, INFN-LNL, Italy ICNMTA2002

At present, four accelerators are in operation at the LNL :                                    Mainly used for solid state physics and applied physics experiments.    Electrostatic accelerator, Van de Graaff type. Single stage-Belt charging system.    Maximum terminal working voltage 2.5 MV.    Available accelerated ions: 1H, 4He single charged    Continuous beams.    Maximum Ion Energy: 2.5 MeV    1 experimental hall; 5 beam lines among which one dedicated to a focussed m-beam facility.   AN2000                                     Mainly used for interdisciplinary research, neutron physics research and advanced educational purposes.     Electrostatic accelerator, Van de Graaff type. Single stage - Belt charging system.    Maximum terminal working voltage: 7 MV.    Available accelerated ions:  1,2H, 3He, 4He single and double charged; 15N double charged    Continuous and pulsed beams.    Maximum Ion Energy: 7 MeV for single charged; 14 MeV for He++; 8 MeV for 15N++    1 experimental hall; 7 beam lines among which one dedicated to radiobiology and collimated m-beam facilities.   CN                                    Used for fundamental heavy-ions nuclear physics experiments and Interdisciplinary researches. At present, Injector of ALPI INFN-LNL: European Large Scale Facility within FP4, FP5,FP6     Electrostatic accelerator, Van de Graaff type. Double stage - Laddertron charging system.     Maximum working voltage: 15 MV.     Available accelerated ions range from 1H to 197Au.      Continuous and pulsed beams.     Energy:from 30 MeV/amu for 1H to about 1.5 MeV/amu for 197Au.     3 experimental halls; 10 beam lines among which one dedicated to radiobiology Tandem-XTU                                 Used for fundamental heavy-ions nuclear physics experiments at intermediate energy and Interdisciplinary researches    Linear superconducting quarter-wave resonant cavities accelerator. At present ions are injected by Tandem    Superconducting material: Lead and Niobium.    Accelerated Ions: 12C and from 28Si to 197Au.    Energy: about 20 MV x ion charge state.    The total output energy of ions from the Tandem-Alpi complex is the sum of Tandem injection energy and ALPI energy: about 35 x ion charge state (MeV).    Experimental halls and beam lines: the same ones of Tandem-XTU. ALPI Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

Available INFN-LNL facilities for radiobiological studies: Light ion broad beam irradiation facility at the 7MV Van de Graaff CN electrostatic accelerator (protons, deuterons, helium-3 and helium-4 ions; E: 0.8-12 MeV) Heavy ion broad beam facility at the Tandem-ALPI accelerator complex (A>4; E: 5-26 MeV/amu) Light ion microbeam facility at the 7MV Van de Graaff CN electrostatic accelerator for “single-ion single-cell” irradiation Fully equipped cell biology Laboratory in near future: Heavy ion microbeam facility at the Tandem-ALPI accelerator complex for “single-ion single-cell” irradiation (A: 6-20; E: few-20 MeV/nucleon) Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

beam collimator assembly thin window supporting flange Ion Beam glass roundel beam collimator gold scatterer vernier scale CHAMBER #1 base plate beam stopper CHAMBER #2 top plate on-line beam monitor beam monitor assembly beam collimator assembly thin window supporting flange Petridish holder especially designed Petridish air inlet aluminized mylar foil turning beam monitor table beam collimators lemo connectors Flanged tube support Teflon gasket Thin mylar foil Stainless steel coupling cap PETRIDISH INFN-LNL Broad-Beam Irradiation facility for Radiobiology Studies Set up at the 7MV CN Van de Graff accelerator and at the Tandem-ALPI accelerator complex Ref: Belli, Cherubini et al., NIM A256(1987)576-580 Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

set up at the 7MV CN Van de Graaff accelerator Schematic view of the INFN-LNL Single-ion microbeam facility for single cell irradiation set up at the 7MV CN Van de Graaff accelerator Silvia Gerardi, INFN-LNL Micro-positioning stages Movable table Tilting Flange with Micro-collimator Biological sample holder Inverted phase contrast optical microscope Beam extraction window Beam-pipe Horizontal ion beam Cooled-CCD - Particle Spatial distribution Monitor Helicoidal guide Single-Ion Detector ( Gerardi, et al., Radiat. Res. 164(2005)586-590 ; 161(2004)93 ; 158(2002)371 ) Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

Especially designed Petri dish Cell holder: Especially designed Petri dish Ext. Diam.: 100mm Int. Diam.: 75mm Cell chamber thickn.: 20m Wet and sterile conditions Ion Beam 7mm mylar Culture medium Mammalian cell ( Gerardi, et al., Radiat. Res. 164(2005)586-590 ; 161(2004)93 ; 158(2002)371 )  in addition: …”intelligent” patterned mylar Petridish under development in a joint collaboration with the Uppsala Group Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

…”intelligent” patterned mylar Petridish under development within a joint collaboration with the Uppsala Group GOLD GRID PATTERNED MYLAR With numbered grid-cells (see: Gunjana Sharma, Uppsala Group) Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

Single Particle Detection Ion beam Microcollimator Petri dish 20um thick cell room Silicon detector Electrostatic deflector Vacuum-air window Trigger signal Active detection silicon detector  spectrometer  deflector trigger Passive detection track detector (CR39)  ion impact points  overlapping with cell map Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006 Silvia Gerardi, INFN-LNL, Italy ICNMTA2002

Ion beam monitoring and spectrometry during cell irradiation Petri dish 20um thick cell room Silicon detector Trigger signal Microcollimator Vacuum-air window Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

Single-ion cell irradiation protocol Cell and Ion position data Particle detector Trigger signal Cell co-ordinates Semi-automatic cell recognition and fiducial marks Image Analysis System and micropositioning stages controller Single ion hit position by cooled CCD Micropositioning stages and Petri dish holder Electrostatic deflector Single Ion data Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006 Silvia Gerardi, INFN-LNL, Italy ICNMTA2002

Cell visualization, micropositioning and revisiting system video Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006 Silvia Gerardi, INFN-LNL, Italy ICNMTA2002

INFN-LNL Radiobiology Laboratory Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

Main elements of the LNL single-ion microbeam apparatus ( Gerardi, et al., Radiat. Res. 164(2005)586-590 ; 161(2004)93 ; 158(2002)371 ) Different light ions (1H+, 2H+, 3He++, 4He++) and enegies available (0.8 – 14MeV, in air) Pinhole Microcollimator in air (and its alignment system) A fast beam deflection system: electrostatic deflector Particle detectors: - high efficiency single-ion counter - spatial particle in air distribution monitor - track detector Semi-automatic Cell visualization and localization system Automatic Cell micropositioning and revisiting system Especially designed Petri dish Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006 Silvia Gerardi, INFN-LNL, Italy ICNMTA2002

….peculiar characteristics ( Gerardi, et al., Radiat. Res. 164(2005)586-590 ; 161(2004)93 ; 158(2002)371 ) Inverted phase contrast optical microscope Software for cell image acquisition and coordinates logging (semi-automatically)  NO FLUORESCENT CELL STAINING  NO UV LIGHT Precision micropositioning stages (Physik Instrumente, D): 0.1m positioning resolution and unidirectional repeatability 0.1m minimum step No backlash Overall positionning precision under microscope: ≤ 2 m Pinhole beam microcollimation in air: 2m or 5m in diam. Counting rate: < 1 ion / second Variety of particles available; Wide LET (and energy) range covered (protons, deuterons, helium-3, helium-4; 7 to 180 keV/mm) Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006 Silvia Gerardi, INFN-LNL, Italy ICNMTA2002

…..Perspectives… Radiobiological studies (cell survival, micronuclei induction, chromosomal aberrations, DNA damage, protein expression) … “bystander effect” ..involving a CELLION Post-doc (biologist; 12 months, since Sept 2005) Improvement/development of an “automatic system” for cell visualization and localization ..involving a CELLION PhD student (Computing Science; 18 months, since Jan 2006) Development, in close collaboration with the Uppsala Group, “patterned mylar Petridish” …twin microbeam facility for heavy ions (6A20) at TANDEM-ALPI accelerator Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

Physics and technological development R.C. , INFN – LNL ….people involved … Physics and technological development R.C. , INFN – LNL Silvia Gerardi, INFN – LNL, in charge for LNL microbeam facilities R&D Giuseppe Galeazzi, INFN – LNL & Padova Univ. Dimitri Zafiropoulos, INFN – LNL Biological aspects: Chiara Zilio, INFN – LNL CELLION Fellows at LNL: Denis Guryev, Russia – ER/Post-doc, Biologist Marcin Skoczylas, Poland – ESR/PhD Student, Computing Science Technical drawings: Marco Rigato, INFN – LNL INFN – LNL CN VdG Accelerator operation staff INFN – LNL mechanical and electronic workshops staff Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

Thanks Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

Extra Slides Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

…related references: S. Gerardi, G. Galeazzi, R. Cherubini A Microcollimated Ion Beam Facility for Investigations of the Effects of Low-dose radiation Radiation Research 164(2005)586-590 “Progress Report of the single ion microbeam facility at INFN-LNL” Radiation Research 161(2004)93-94 R. Cherubini, M. Conzato, G. Galeazzi, S. Gerardi “Light ion micro-collimated beam facility for single ion - single mammalian cell irradiation studies at LNL-INFN” Radiation Research 158(2002)371-372 M. Conzato, R. Cherubini, G. Galeazzi, S. Gerardi, L. Read “Light ion micro-collimated beam facility for single ion – single mammalian cell irradiation studies” LNL Annual Report 2000, INFN-LNL (REP) 178/2001, p. 90-91 R. Cherubini, G. Galeazzi, S. Gerardi “Development of a cell recognition, positioning and revisiting system for radiobiological studies with low-energy single ions” LNL Annual Report 2001, INFN-LNL (REP) 182/2002, p.81-82 Silvia Gerardi, Giuseppe Galeazzi, Roberto Cherubini “Characterization of microcollimated beams in air delivered by the single ion microbeam facility at the 7MV CN accelerator” LNL Annual Report 2002, INFN-LNL (REP) 198/2003, 63-64 Silvia Gerardi, Eugenia Tonini “CELLView: a software control system for sample movement, single-cell visualization and micropositioning at the LNL horizontal single-ion microbeam facility” LNL Annual Report 2002, INFN-LNL (REP) 198/2003, 65 Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

In air ion beam extraction flange and microcollimator alignment system Technical drawing: lateral section Photo: front view Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

Beam monitor and single ion spatial distribution in air Custom-made cooled CCD camera: - no optical lens - no shutter - 3.0 mm Havar window - pixel : 6.8 x 6.8 mm2 - array : 2184 x 1472 Ultrathin window for particle detection Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

Electrostatic beam deflector Plate A 3V0 (Volt) Wires 1-1 2V0 (Volt) Wires 2-2 V0 (Volt) Wires 3-3 0 Wires 4-4 - V0 (Volt) Wires 5-5 -2V0 (Volt) Plate B -3V0 (Volt) V0 = 200 V, during deflection stage Response time = 150 ns beamline Ion beam Trigger signal: Beam on / Beam off (Bizzetti et al., NIM 159 (1979) 575-579) Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006