Olympus Fluoview FV1000
fv 1000
Switching On
1)
Switch on the two black rocker switches on the Power Manager units
Rocker
switches
power manager unit
2)
Check HBO mercury lamp is on. Switch on green rocker if off.
Green rocker
Olympus Fluoview FV1000
HBO
3)
Check visible lasers are switched on (left of the system table). Turn key from 12 to 3 o’clock
if not already on.
Laser key
Visible lasers
4)
Check 561 Diode laser is on.(room 125) If no orange lights displayed press laser on green
button.
Orange lights
Green button
561 laser control unit
5)
Switch on the computer.
Olympus Fluoview FV1000
6)
7)
8)
9)
Log onto the computer as normal
Double click on the FV10-ASW.1.7 software icon on the desktop
Select user ID | User
Select password | xxxxxx| click on OK | Wait until all windows are open correctly.
10
Select objective lens from Acquisition Settings x10 | x20 | x40 dry lenses | X60 oil
The lens will change automatically to the lens you choose in the software. This can also be
selected from the microscope control box .
Olympus Fluoview FV1000
Microscope
Control
Box
Objective
lens
11
Set up your sample on the microscope. Make sure the shutter is open on the mercury bulb
shutter
12
Check sample with epi-fluorescence/ bright field
Olympus Fluoview FV1000
Click here for Trans
Click here for epi lamp
BF Lamp
Intensity
13
For epi-fluorescence choose the appropriate filter.(Microscope control box at right of
microscope on the air Table )
NUA | DAPI
WU | DAPI LP
NB | FITC/GFP
WG | TRITC/TR
Setting UP software for XY scanning.
1
Use the explorer window to create a folder to save images eg: Olympus_date
Create two folders inside this folder eg: Date_Tif | Date_oib
The computer is networked
2
From the Image Acquisition Controls select dyes from the dye list.
Olympus Fluoview FV1000
To select a dye click here
To remove dyes double click here
Select required dye from list
Select Apply
After your first session on the confocal the settings can be loaded from saved oib files
In the explorer window right hand mouse click on the oib file and choose load
Acquisition parameters
Select VBF from the Image Acqusition Controls to set emission to be collected
Set up all other acquisition settings
The FV1000 has 6 laser lines 405,458,488,515,561 and 633.
3
4
5
VBR
4
To obtain an image
Click on XY Repeat |if nothing appears on the screen | Adjust HV | Adjust pinhole and focus
to see an image.
Olympus Fluoview FV1000
Faster speeds
for
Setting up
image
Click Stop once image is set up
Click here to capture
5
6
7
Save image as oib file . Can be save as tif
For more than 1 channel and to avoid bleed through tick the Sequential scan box
For removal of noise from the background tick Kalman | line filter | Select 2 to 6
Olympus Fluoview FV1000
8
In the live view window right hand mouse click and select add a view. Make sure show
overlay is ticked from the drop down menu. This will then allow you to see an overlay when
scanning.
Setting up the software for XYZ scanning
1
2
Set up image as before | Select Depth from the image acquisition controls
Set up Z positions using the Z scan controls from the Acquisition settings window
Click set to select top and bottom
Fine focus
3
4
Select depth from Acquisition control window
Select XY to start scan
Olympus Fluoview FV1000
5
Series done shows up. Click to accept
Save as oib files. Can also do projection and save as tif.
Switching Off
1
2
Remove slide and clean lens
Exit the software and shutdown the computer
3
Turn the Visible laser key from 3 to 12 o’clock allow to cool 5min.
4
5
Clean up microscope area.
Switch off the two black rocker switches on the Power Manager unit.