Laboratory for Fluorescence Dynamics

A NIH research center for biomedical fluorescence spectroscopy at the University of California, Irvine

LFD User Guide

Last updated May 10, 2006.

Assistance

The LFD staff members listed below are available to assist all users with work in the LFD.

  • The Director of Operations oversees the instrumentation and assures timely scheduling of instrumentation.
  • The User Coordinator, is responsible for instrument scheduling and can assist in experimental design and in helping users to carry out their fluorescence measurements. She also helps in providing the needed biochemical and chemical materials necessary for sample manipulation and is available to assist in sample preparation.
  • Milka Stakic, Laboratory Assistant, assists with incoming shipments, supplies, and cell cultures.
  • Christoph Gohlke, the System and Network Administrator, assits in networking user computers.
  • LFD graduate students are well versed in both the instrumentation and analysis, and would be happy to assist you when possible.

User Procedures

Space

Users of the LFD for a medium to long length (more than 1 week) will be provided desk space.

Communication, copying, and computers

Because the University of California and the LFD operates on a strict, limited budget, we request that all outgoing long-distance calls be kept to a minimum. A public telefax is available in the Biomedical Engineering Department business office (room 3120). The telefax number is 949-824-1727. All outgoing faxes should be charged to your telephone credit card. The telefax should be used only in an extreme emergency. If telephone or othercharges are great, it may be necessary to charge them back to the user. Computers are available on a first-come, first-served basis in thecomputer room. Each is equipped with all software for data analysis, word processing, networking, etc. Secretarial services are available only as they relate to core research collaborations.

Keys

Due to university policies, no keys can be issued to users of the LFD facilities.

Reserving time on the instruments

Before preparing samples or planning experiments, discuss your planned experiment and your instrument and biochemistry needs with Dr. Sanchez. She will schedule time for you on the necessary instrument(s) and will make sure that the instrument(s) is correctly configured for your needs.

Reagents

Generally, we do not supply reagents for visiting researchers.

Using the instrumentation

The first time you use an instrument, you will be given instruction on how it functions. LFD personnel will be on hand to help (during work hours), in case of problems. Please excuse various inconveniences due to hardware and software updates. We try to incorporate new developments as smoothly as possible, but of course, these things do not always go as planned. Please also remember that lamps, lasers, tubes, lenses, etc. have finite lifetimes and that they may "die" just when you need them. We will do the best we can to ensure that your visit goes smoothly.

Evenings/weekends

LFD staff will generally be available for help after work hours, but also have finite lifetimes. Assistance cannot be assured after 5:00 PM and on weekends.

Paperwork

Before you end your visit at the LFD, please complete the online service form at http://www.lfd.uci.edu/service/form/. Most importantly, please provide a title and abstract of your work performed at the LFD. Typically, there are no fees charged for using the LFD facility. Information from the user forms is used for our yearly progress reports to the NIH and for our grant renewal justification.

Data storage

All data files, except uncompressed video data, acquired during experiments must be saved on the data file server. All data must remain in the LFD when you leave.

Uncompressed video data, such as those acquired during TIRF, must not be saved on the file servers. Users performing such experiments are required to provide their own external USB hard drive for data storage. One day of measurements typically generates 10-50 GB of uncompressible video data.

Data analysis

We have software for data analysis and are happy to help you use it, discuss your results with you, and perhaps suggest further studies. Please feel free to talk with any of the professional staff about your results.

Publications

If the results obtained at the LFD are included in a publication, we request that you acknowledge the LFD at the end of the manuscript with the following statement:

"The experiments were performed at the Laboratory for Fluorescence Dynamics (LFD) at the University of California, Irvine (UCI). The LFD is supported jointly by the Division of Research Resources of the National Institutes of Health (PHS 5 P41-RRO3155)and UCI."

If you have worked closely with staff of the LFD and they have contributed significantly to your results, you may wish to include their name as a coauthor of a paper you produce as a result of your work in the LFD. This is not mandatory. If you choose to include a LFD staff member as coauthor, please provide them with a draft copy of the manuscript you plan to submit.

Problems

If you have any problems with data acquisition, please notify Susana Sanchez, who will attempt to rectify the situation.

Laboratory Safety

For all emergency situations dial

as appropriate. The LFD is located in the third floor of the Natural Sciences II building room 3311.

A medical kit is located in a marked drawer left of the fume hood in the main biochemistry laboratory.

Fire Extinguishers can be found by the doors of the biochemistry laboratory and the spectroscopy laboratory.

Chemical Material Safety Data Sheets are located in the biochemistry laboratory.

Laser Safety Manuals and Laser Specification Manuals are also located in the biochemistry laboratory.

Do not work alone in the Lab!

Laser emission hazards

LFD-available lasers:

  • 2 Nd:YAG Lasers
  • 2 Dye Lasers (pumped by the Nd:YAG Lasers)
  • 1 Argon Ion Laser
  • 1 Helium Neon Laser
  • 1 Titanium Sapphire Laser

The Nd:YAG fundamental frequency, 1064 nm, is invisible to the eye, but can cause severe eye damage. The 1064 nm beam is the most powerful (25 Watts) at the LFD. This beam is primarily confined to the laser head but will be co propagated with the visible line, 532 nm, produced from SHG (second harmonic generator) equipped lasers (both Nd:YAG lasers). If the internal beam dump is moved, the 1064 nm line will no longer be confined, thus laser safety glasses blocking such light should be worn.

The visible lines (direct or reflected) from the Nd:YAG (532 nm) or from the pumped dye lasers (depending on the laser dye being used) should be shielded from the eye with safety laser goggles.

Doubling of pumped dye lasers typically produces wavelengths between 280 nm and 410 nm which are invisible to the eye extremely hazardous.

Hazards from the continuous wave Argon Ion (Model 2025) laser lines includes both visible (ranging from 462-528 nm) or invisible (351 & 364 nm) radiation, depending on the selected line, while the Helium-Neon laser produces 632 nm radiation at 35 mW of power. All these laser emissions (or reflected beams) can do permanent eye damage.

No users will align, tune, or modify a laser without proper instruction from the LFD personnel.

Eye damage

Eye contact with laser light can result in eye damage or blindness. This is true for the direct laser output beams as well as the scattered beams off polished surfaces (mirrors, beam splitters, lenses, etc.). Some laser lines are invisible to the eye and not easily detected.

General guidelines

  • Do not stare at laser light reflected from any surface.
  • Never stare at the laser output ports.
  • Close the protective curtains so spurious beams are blocked from the rest of the laboratory.
  • Remove all reflective objects from your wrist sand hands (i.e., rings, watches, ) when working with the laser.
  • Do not wear loose neck scarves or neckties which may drape into the laser light and reflect the beam.
  • Do not put solutions on top of the laser or laser power supply. Flasks can be easily knocked over and create a shock hazard (this includes drinks, which must not be present in the first place).
  • Do not put hands near electrical components of the laser.
  • Do not remove protective covers, beam stops or barriers.

Fire and burns

Laser beams are powerful enough to burn skin or to ignite volatile substances such as alcohol, ether, gasoline and other solvents. Be aware of the laser light paths and avoid placing flammable substances, or your arm, in the beam path.

Electrical shock

Though most of the high voltage areas (lethal voltage) of the laser are protected, the laser covers are occasionally removed for laser repair and servicing. In these instances all circuit boards and every part of the electrical system (including laser head) should be treated as high voltage areas.

Biochemical hazards

Radioactivity

Use of radioactive chemicals is not permitted at the LFD.

General Lab Procedures

First and foremost be considerate and use common sense. Others may not be familiar with the particular hazards associated with your experiments.

Work areas should be limited, i.e., don't drag your reagents and chemicals to all parts of the laboratory. Areas near commonly used equipment, the spectrometer, balances, FPLC or the lyopholizer, should be cleaned immediately after use. Any solutions that you make must be labeled with the name of the user, the contents and the date. Return all chemicals to their proper place after you have finished with them.

The LFD user coordinator should be notified if any hazardous chemicals are brought into the laboratory. If you are unsure of the potential hazard of a specific reagent consult Material Safety Data Sheets (located in the biochemistry laboratory) or consult one of the LFD personnel.

Skin/eye contact with hazardous or noxious chemicals

  • Use gloves, lab coats, and face/eye protection(contact lenses are discouraged) as appropriate for the particular chemical (acids,organics, etc.) with which you are working. Remove gloves before leaving your work area to not spread chemicals to door knobs, pencils, pens, food, etc.
  • Keep hazardous chemicals in a controlled and restricted area to avoid contamination of the laboratory.
  • Discard any cracked or broken glassware in the appropriate refuse container.

Ingestion and inhalation

  • Food and drinks are not to be taken or consumed in the laboratory.
  • Application of cosmetics is not permitted in the laboratory.
  • Mouth pipetting is forbidden.
  • Inhalation hazards from volatile solvents,aerosol or dust-producing chemicals/solutions should be minimized by working under a fully ventilated fume hood (and with particle masks, if appropriate). Non hazardous, nuisance odors, such as b-mercaptoethanol, should also be used in the hood since vapors escaping to the hallways might be misinterpreted as a gas leak.

Hazardous spills

  • Use absorbent paper under your work area if hazardous materials are used and do not crowd containers, or place them where they could be easily knocked over.
  • If a hazardous spill does occur, notify LFD personnel, call 911 for medical police, and fire.
  • A spill containment kit is located under the table top centrifuge in the biochemistry laboratory.

Fires

  • Smoking is not permitted in the LFD for fire,health and equipment reasons.
  • Fire extinguishers are located next to the doorways of the spectroscopy laboratory and the biochemistry laboratory.