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Laser Safety Laser Safety Ronald Breedijk LCAM safety talk Laser Components • Light Amplification by Stimulated Emission of Radiation OPTICAL RESONATOR LASER beam Active medium high reflectance output coupler mirror mirror excitation Associated hazards: 1. Laser Beam: eye injury, burns, skin cancer (UV), fire hazard 2. Excitation source: high voltage, water cooling Ordinary Light vs. Laser Light 1. Many wavelengths 1. Monochromatic 2. Multidirectional 2. Directional 3. Incoherent 3. Coherent These three properties of laser light are what can make it more hazardous than ordinary light. Laser light can deposit a lot of energy within a small area. LASER SPECTRUM Gamma Rays X-Rays Ultra- Visible Infrared Micro- Radar TV Radio violet waves waves waves waves 10-13 10-12 10-11 10-10 10-9 10-8 10-7 10-6 10-5 10-4 10-3 10-2 10-1 1 10 102 Wavelength (m) LASERS Retinal Hazard Region Ultraviolet Visible Near Infrared Far Infrared 200 300 400 500 600 700 800 900 1000 1100 1200 1300 1400 1500 10600 Wavelength (nm) Laser-Professionals.com Setup LSM 510 Argon UV laser – 351nm and 365nm Diode laser – 440nm Argon laser – 488nm and 514nm Hene laser – 543nm DPSS laser – 561nm Hene laser – 633nm Ti:Sapphire laser – 700nm – 1000nm (two photon) SetupA1 excitation Nikon options A1 4 laser box Transmission 640 nm 561 nm 457 nm 405 nm Diode Dpss 488 nm Mercury diode 514 nm AOTF Objectives 1. Plan Apo VC 60x, NA1.40 (oil) 2. Plan Fluor 40x, NA1.3 (oil) 3. Plan Fluor 20x, NA0.75 (M im.) Main dichroic 1. 405/488 sample 488 3 laser 2. 405/488/561 440 594 3. 405/488/561/640 514 box Mercury lamp cubes nm nm 4. 457/514 nm 5. 20/80 1. BFP 2. CFP 3. GFP 4. YFP Scanner 5. RFP 6. Empty AOM Beam Combiner 1. Open (only 4 laserbox) Beam * 2. Mirror (only 3 laserbox) combiner 3. 50/50 Human Eye - laser beam can be focused by cornea and the lens to a very tight spot on the retina 400-1400 nm 300-400 nm Retinal damage Types of laser eye exposure Laser hazard classes • Classification by wavelength and output power, according to their ability to produce damage Class Power Remarks Typical examples I Very low •Inherently safe, CD, DVD drives, laser or beam •No possibility of exposure printers… completely enclosed II 1 mW •Staring into the beam is hazardous Supermarket laser Visible only •Eye protected by aversion response scanners, some pointers IIIa 1-5 mW •Aversion may not be adequate Laser pointers IIIb 5-500 mW •Direct exposure is a hazard Ar / Kr lasers Diode /DPSS lasers HeNe laser IV >500 mW •Exposure to direct beam and scattered Two photon laser light is eye and skin hazard OPSL lasers •Fire hazard Safety measures • Be informed • Eyewear for classes IIIb, IV for everybody in the room • Beam paths above >200 mW should be guided through tubes • Highest risk during alignment, optical setup modification • Laser warning signs Common practice How to handle objectives • Immersion • Stage movement • Z-control • Change sample -> park objective Different objective types After using the microscope • Remove sample safely • Clean objective • Clean microscope surrounding (oil stains, lens paper, etc.) • If you find the microscope not cleaned notify the staff Cleaning objective • Remove immersion with lens paper • If neccesary clean with Ethanol • Have a close look Consumables Rules and guidelines for guests of the Van Leeuwenhoek Centre for Advanced Microscopy (LCAM) General rules • LCAM can be used by students, UVA personnel and external users. The projects needs to be approved by the LCAM staff. • No modifications to the instruments • Clean the equipment after each session. • Switch off the equipment. In case not: fine. • Fault messages must be recorded in each instrument’s logbook. • Immediately report instrument damages to the LCAM staff. • For the use of fotons the user will be charged. The amount at this moment is set at €7,50 per hour Safety, Chemicals & Biohazards • No food and beverages in the microscope rooms and laboratories. • In case of a toxic / dangerous chemical users must notify the managing staff. • No radioactive materials. • Potential biohazard material must be cleared with the staff • Live cells, media, Petri dishes and gloves must be discarded in the biohazard container in the tissue culture room. No bacteria! • Clean the equipment at the end of the experiment. • The microscopy labs are ML1 labs. For ML1 work fill out the GMO-form and hand it over to Joachim. - Failure to notify the staff of chemical or biological hazards will result in immediate exclusion from CAM Time Use Policy • The equipment has to be reserved in advance. • No reservation more than 1.5 weeks in advance. • Reserve by sending a email to [email protected]. • The schedule is made on Thursday afternoon. • The schedule can be viewed at: www.lcam-fnwi.nl/booking/ • The staff can break into the schedule (i.e. maintenance). • Only guests with UvA permission may get LCAM after- hours permission. • After-hours use must be reserved. • Unauthorized after-hours use of LCAM equipment will result in exclusion from the facility. Data Storage and Computer Use • Don’t store data on the local computer. • Each user will be allocated 20 GB storage space. • Only available for storage of LCAM data. • We strongly recommend not to rely exclusively on the server for data safekeeping! Users are responsible for their own data. • Internet browsing, reading email or playing music is not allowed on our computers. Publications • LCAM is not a facility; it is an expertise centre. This implies that the LCAM-staff works together with the guests of LCAM based on a scientific collaboration. • LCAM should appropriately be acknowledged in publications. LCAM should be cited as: ”Van Leeuwenhoek Centre for Advanced Microscopy, Section Molecular Cytology, Swammerdam Institute for Life Sciences, University of Amsterdam”. • LCAM wants to receive a pdf-file of the publications. • Note: For grant applications (e.g. new equipment) a collaborative papers are the only prove for collaboration between research groups. Training • Before a researcher starts as a guest of LCAM, plans will be discussed with the LCAM-staff. Also matters like financing, use of equipment, best research strategy will be discussed during this intake interview. • Before a researcher starts as a guest of LCAM, a project description sheet will be filled in together with the LCAM- staff. • Training by one of the LCAM staff members is required before using any LCAM equipment. • On regular basis, LCAM will organize 1-day training for new guest researchers. Users of LCAM should attend such course and pass the exam. Information slide Questions? .
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