MENU
Olympus banner

Olympus FV1000

Tell a friend about this page.
All fields required.
Can be sent to only one email address at a time.
Share Facebook Icon Twitter Icon

Olympus FV1000 Laser Scanning Confocal & Multiphoton Microscope

The Olympus Fluoview FV1000 confocal laser scanning microscope is a powerful tool for high resolution multi-dimensional observation of cell and tissue morphology, and precise molecular localization. In addition to confocal microscopy, this system is also capable of multiphoton imaging as well.

The FV1000 system is equipped with a motorized stage controller, heated CO2 incubation system, 6 confocal laser lines, tunable multiphoton laser, 4 objectives, 3 epifluorescence filter cubes, confocal and multiphoton detection system, and many other great features.

Location

The system is located on the fourth floor of the East Carolina Heart Institute at 115 Heart Drive in room 4305. You will need ECU one card access for the fourth floor to gain access to this space. The system is available for all ECU employees to use.

Contact Jim Aloor, PhD for new training or questions while using the system.

  • Location of the system: ECHI 4305
  • Phone in ECHI 4305: 252-737-5153
  • Dr. Aloor's office: 252-744-0404

Cost

  • $30 per hour to use during peak times (8:00 AM - 5:00 PM)
  • $10 per hour to reserve peak times (8:00 AM - 5:00 PM). This amount is refunded for time used, i.e. only $20 per hour total to use the system with a reservation
  • 50% discount for off-peak times (5:00 PM - 8:00 AM). Only approved users are allowed to use the system during off-peak times.
  • $60 per hour for assisted use during peak times.

These charges will be evaluated on a quarterly basis.

About the system

Olympus Flyers

Objectives

The Olympus IX-81 inverted microscope can hold a total of 6 objectives at one time. We currently have 4 installed.

Epifluorescence

Fluorescence can be observed on through IX-81 using the mercury lamp and filter cubes. We currently have 3 filter cubes installed.

  • DAPI (Hoechst/AMCA): ex 335-375, em 440-500
  • FITC (EGFP/Bodipy/Fluo3/Di O): ex 465-495, em 515-555
  • WIGA (TRITC, Di I): ex 515-550, em 600-640

Explanation of microscopy via Epi-fluorscence illumination and common filter cube combinations

Confocal Lasers

Fluorescence can be observed on through confocal laser scanning. We currently have 6 laser excitation wavelengths for confocal imaging.

  • 405 nm
  • 458 nm
  • 488 nm
  • 515 nm
  • 559 nm
  • 635 nm

Fundamental concepts of confocal microscopy

Confocal laser scanning

The FV1000 system has three photomultiplier tubes for confocal detection and one transmitted light. This setup allows for three fluorescent dyes to be imaged simultaneously along with another channel for transmitted light or DIC imaging. In addition, you can add up to 12 additional channels by using the virtual channel selection in the software.

Multiphoton Laser

The FV1000 system has a Spectra-Physics Mai Tai Ti:Sapphire multiphoton laser. This laser is attached to a chilling system located underneath the air table and should NEVER be turned off. The detection system is a non-descan detector, separate from the multiphoton channels. Two multiphoton channels are available, adjusted by changing the filter cubes.

Non-descan detector filter cubes

  • FV10-MRV/G - Violet/Green - em 420-460 nm,  495-540 nm
  • FV10-MRC/Y - Cyan/Yellow - em 460-500 nm, 520-560 nm
  • FV10-MRG/R - Green/Red - em 495-540 nm, 575-630 nm

Multiphoton tutorial

User manuals

Operating Instructions

Important Precautions

  • Mercury Lamp: Once you turn the lamp on, it needs STAY on for at least 30 minutes before being turned off. Once it’s turned off, it needs to be off for at least 15 minutes before being turned on again. The mercury lamp has a 300 hr bulb; if the counter passes 300 hrs—do not turn it on—the bulb needs to be changed.
  • Lasers: Turn all lasers you’ll need during initial startup. Let the lasers warm up 15 min before use, and cool down after use before power cycling. Do not turn lasers on mid-run!
  • Check the signup calendar: If someone is signed up to use the system within an hour of when you finish, leave the system on for this user. However, you can log out of the software. If you are the later user and you decide to cancel your experiment, it is your responsibility to check to make sure the system is powered down.
  • Oil immersion: There are three types of objectives: air, oil, and silicon oil. Do not mix the oils, and don't get oil on the air objectives. Only a very small amount of fluid is required. Silicon oil is the green bottle, regular oil is the blue bottle (or small clear bottles). If you’ve put oil on a slide, be careful about changing objectives. When finished using immersion lenses, clean the objective using lens paper and cleaning fluid, never Kimwipes!. Guide to keeping a clean microscope.

System startup

Follow the instruction cards hanging in front of each item on the rack. The green numbers are the turn on order

  1. Fill out the logbook. We use this to keep track of usage of the microscope lasers, mercury lamp, and objectives.
  2. Take off the microscope cover.
  3. Mercury burner power supply (if needed), Switch power on.
  4. Multiline Argon laser 458nm, 488nm, 515nm (if needed), Switch power on then turn key on.
  5. 559nm laser diode (if needed), Switch power on, but wait to turn the key until TEMP light stops blinking. Turn key on once temp light is solid.
  6. Multiphoton power supply (if needed). Switch power on.
  7. Laser combiner power supply and 405nm and 635nm laser diode. Switch power on. Must be on for any laser function
  8. Microscope control box. Switch power on
  9. Microscope power supply. Switch power on. Switch key on (if not already). Key can be left on all the time.
  10. Computer. Right power button.
  11. Log into ECU network, then Log into BookIt Lab tracking software.

System shutdown

Follow the instruction cards hanging in front of each item on the rack. The red numbers are the turn off order

  1. Turn off laser keys to allow the lasers to cool. Power off multiphoton if it was used.
  2. Check BookIt Lab site for other users.
  3. Cleanup oil, objectives, and any samples.
  4. Load data off computer and shut computer down.
  5. Microscope power supply. Switch power off. Switch key on (if not already). Key can be left on all the time.
  6. Microscope control box. Switch power off
  7. Laser combiner power supply and 405nm and 635nm laser diode. Switch power off.
  8. Multiphoton power supply (if used). Switch power off.
  9. 559nm laser diode (if used), Switch power off. Key should already be off.
  10. Multiline Argon laser 458nm, 488nm, 515nm (if used), Switch power off. Key should already be off.
  11. Mercury burner power supply (if used), Switch power off.
  12. Cover the microscope.