1. 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

2. 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

3. 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: 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

4. 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)
Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

5. 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) ; 161(2004)93 ; 158(2002)371 )
Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006

6. 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) ; 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

7. …”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

8. 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

9. 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

10. 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

11. 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

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

13. Main elements of the LNL single-ion microbeam apparatus
( Gerardi, et al., Radiat. Res. 164(2005) ; 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

14. ….peculiar characteristics
( Gerardi, et al., Radiat. Res. 164(2005) ; 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

15. …..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

16. 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

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

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

19. …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)
“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)
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
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

20. 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

21. 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

22. Electrostatic beam deflector
Plate A 3V0 (Volt)
Wires V0 (Volt)
Wires V0 (Volt)
Wires
Wires V0 (Volt)
Wires V0 (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) )
Roberto CHERUBINI, INFN-LNL - CELLION Mid-term Meeting - CENBG, Bordeaux,Feb 3-5, 2006