Olympus Fluoview FV1000 Handbuch

Olympus FV1000 -1- Short instructions

Short instructions. Olympus FV1000

Version 110608

This short instruction will guide you through the turn on and off sequence of the microscope

system. For a detailed instruction in system operation refer to the Olympus user manual.

This instruction also contains important information on laser safety and general guidelines.

Content:

1. Lasersafety Page2

2. General guidelines Page 3

3. Switching on the microscope Page 5

4. Using the climatic chamber Page 6

5. Switching off the microscope Page 7

Olympus FV1000 -2- Short instructions

1. Laser safety

Safety notes for operating the laser scanning microscope:

During scanning, the laser radiation is freely accessible after exiting the objective in

the specimen area of the laser scanning microscope. This circumstance demands

special attention and caution. If the laser radiation comes in contact with the eyes, it

may cause serious eye injuries. For this reason, prudent handling is absolutely

necessary as soon as one or several laser emission warning indicators are lit. If used as

prescribed and observing the safety notes during the operation of a laser scanning

microscope, there are no dangers to the user.

1. According to IEC/EN 60825−1, this laser scanning microscope is a laser

device of Class 3B.

2. Never look directly into a laser beam or a reflection of the laser beam. Avoid

all contact with the laser beam. Otherwise, your eyesight may be permanently

damaged. A reflected laser beam is just as dangerous as a direct beam.

3. Never deactivate the laser protection devices. Read the “Safety Guide” in the

Olympus User’s Manual to familiarize yourself with the safety devices of the

laser scanning microscope. If you find any of the laser protection devices

inactivated, please contact the facility staff immediately.

4. Note that objects (such as micromanipulators) in the specimen area may cause

laser light to exit the safe beam path during the scanning in an uncontrolled

manner by means of reflection or scattering and endanger the environment.

5. Do not change a specimen during scanning.

6. Do not change any objectives during scanning.

7. All unused positions of the objective turret should be closed with a cap.

8. Do not change any filter cubes or beam splitters during scanning. Never

disconnect an optical waveguide.

9. Never remove the scan head from the microscope stand during operation.

Before removing the scan head, the system must be completely switched off.

10. Do not remove the microscope condenser.

Olympus FV1000 -3- Short instructions

2. General guidelines

1. Lab rules:

This room is a S2 laboratory.

At times when S2 experiments are performed in this room, you are not

allowed to enter the room. S2 experiments will be indicated by a sign at the

outside of the door and in the online booking system.

When no S2 experiments are performed everyone who is authorized by the

facility staff can enter the room.

You always have to wear a lab coat. Lab coats can be left in the cupboard,

first door left from the entrance.

2. Booking and keys:

The use of the Olympus FV 1000 system is restricted to live cell experiments.

Before you can use the microscope you have to make a reservation in the online

booking system. Please note in the reservation the desired temperature. If you

don’t use the climatisation control not this also in the reservation

Always pick up a key from Carsten Lindloff in room 1052. Office hours: Monday

to Thursday, 7:30 – 16:00; Friday 7:30 – 14:30. Phone: -8458. If you are planning

to use a microscope outside these hours, pick up a key in advance.

When you are leaving lock the microscope room and after 16:00 also lock the

main door of the facility. Return the key to Carsten Lindloff or to the mail box of

the Laser Microscopy Facility, level S, in the hallway left from the stair case,

second row, in the center.

3. Climatisation control:

Please note in the online reservation the desired temperature.

Carsten Lindlow will switch on the temperature control early in the morning to

ensure that the incubation chamber has reached the temperature when you start

your experiment.

But you are responsible for setting CO2 and humidity as well as for switching off

the climatisation control. Read chapter

4. Immersion oil:

Always remove old immersion oil, before you add new ones.

Only use the lens tissues provided by the Laser Microscopy Facility staff.

In case there is oil on no-oil objectives, don’t try to clean them but leave a note

in the log book.

Olympus FV1000 -4- Short instructions

5. Changing objectives:

Never change the objectives manually!

To change objectives use the microscope control menu within the Olympus

software or the panel box left of the microscope stand.

6. Changing the specimen:

Before you change the specimen, drive down the objective nosepiece.

Don’t forget to remove the old oil from the objective before you add new ones.

7. Lasers:

Lasers for the lines 491, 532, and 559 nm have to be switched on manually. Read

chapter 3 for details.

8. Data transfer:

There are three ways to transfer your data from the microscope computer:

a. Transfer through the MHH network. First, create a shared folder on the

computer which should receive the data. At the microscope computer, search

for the receiving computer using the IP number. Make an alias on the desktop

for future usage.

b. USB-sticks or movable hard disks connected via USB. The USB sockets are

located at the front of the computer.

c. Writable DVDs. Open the Nero software and follow the instructions.

Olympus FV1000 -5- Short instructions

3. Switching on the microscope

1. Follow the procedure by the numbers:

, , …………..

2 5

2. Switch on the HBO lamp (no. 1, underneath the computer table).

3. Switch on buttons no. 2 and 3. Microscope stand, laser and scan controllers, and the

computer start automatically.

4. Start the Olympus software: double click on “FV1000 1.7. Start”.

5. Select your User ID, type in your password, and press OK.

6. Do not perform the next step until the software start up is completed. If you get any

error messages at this step consult the trouble shooting guide.

7. This microscope provides the following laser lines:

405 nm

440 nm

491 nm

532 nm

559 nm

655 nm

The lasers for 405, 440, and 655 nm are controlled entirely through the Olympus

software. These lasers are switched on by selecting an acquisition protocol from the

dye list in the “Image Acquisition” window or by directly clicking on the laser box in

the “Acquisition Setting” window.

The lasers for 491, 532, and 559 nm have to be switched on manually.

Note, the software will not recognize, if they are not switched on.

Switch on only the lasers you need.

8. To switch on 491 and 532 nm, turn the ignition key (no. 4) to the vertical position. The

LED “LASER ON” will turn green. LED “TEMP LOCK may flash for 60 sec. Then

all four LEDs should be green.

9. To switch on 559 nm (no. 5), wait until the LED 'TEMP' stops blinking. Then turn the

ignition key (no. 5) to the ON position. The red LEDs “Laser” will blink for a few

min. Then they should stay red.

Olympus FV1000 -6- Short instructions

4. Using the climatic chamber

1. Check the water bottle at the control unit behind the microscope table. Refill with

distilled water only. Distilled water is in Lindloff’s office.

Open the valve underneath the water bottle.

2. Turn on the power switch at the right side of the control panel No. .

3. Switch on the microscope following the procedure in chapter 3. The climatisation

control software opens automatically.

4. In the software set the desired temperature between 30° and 42° C. Cooling is not

possible. Do not activate the humidity before the temperature is stable to avoid

condensation which will damage the microscope equipment.

5. If you are using the CO2open the gas bottle at the wall behind the microscope table.

Turn the main valve counter clockwise, not more than a quarter turn.

6. When the temperature is stable set the desired values for CO2concentration (between

0 and 10%) and relative humidity.

Don’t set the relative humidity higher than 60%. Higher values will cause

condensation which will damage the microscope equipment. The software will

open a warning. Confirm by pressing no.

7. For shutting down, select first the dry mode in the software. The values for CO2and

humidity will be set to minimum and the temperature to 37°C. Let the dry mode run

at least for one hour.

8. Close the gas bottle, if used. Close the valve underneath the water bottle.

9. Turn off the power switch at the control panl No. .

Olympus FV1000 -7- Short instructions

5. Switching off the microscope

1. Exit the control software.

2. Turn the key switch of the 491/532 nm laser to off (number 4).

3. Turn the key switch of the 559 nm laser to off (number 5).

4. Switch off numbers 3 and 2.

5. Switch off the HBO lamp (number 1).

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