111813_Nikon SD Training Startup

Nikon Eclipse TE2000 Spinning Disc
Nikon Eclipse TE2000 Spinning Disc
inverted perfect focus microscope
optimized for live cell imaging and time lapse movies
Equipped for DIC, Ludl Piezo Z-stage for 3-D imaging
temperature controlled chamber
widefield cubes for GFP, YFP, TxRed, CFP, DAPI
Cascade 512B and Coolsnap HQ CCD detectors offer
simultaneous data acquisition
• 488 and 568 nm Ar lasers
• Visitech QLC100 confocal head.
Turning on and Setting up the Microscope
1. Turn on Power to the AOTF Controller
2. Turn on Power to the Laser Power Supply
3. Turn on Power to the Laser Supply Remote
a: Flip switch to “operate”
b: Turn key past “on” and let go
c: Laser should go to “100” or more
d: Let warm up 30 min before using
e: Flip switch to “standby” if not
using the lasers for awhile or when
you leave
f: Laser Power can be displayed in
mW or in AMPs depending on which
position the toggle is flipped
g: Laser Power can be changed by
toggling the switch up (increasing
power) or down (decreasing power)
4. Open AOTF Controller Software
- Adjust % Power in software if
- Usually start with 100%
- Must do this before doing next
5. Turn on Black APS Power Supply
6. Turn on Temperature Contoller:
- Turn dial to “set” option and use
dial to set control temperature.
- Keep at 39oC so that the center of
your dish is 37oC.
- Turn dial to T1 to see actual temp.
7. Turn on Ludl Shutter Control:
- Widefield/DIC is Halogen bulb
- Fluorescence is mercury bulb
8. Turn on Xcite Mercury Lamp
9. Turn on Halogen Lamp
*This lamp is under the air table.
10. Turn on confocal head key
*Both lights should be on.
11. Turn on the Perfect Focus Box:
- Push green button to “engage”.
- If green light flashes, it is not
working properly.
- Be sure to “disengage” PF by
pushing green button before
removing your dish of cells from
the stage.
12. Turn on Power to the Microscope:
- Back left bottom of scope
where red tape is located.
13. Turn on Camera(s) before opening IPLAB:
- Cascade is for Confocal
- CoolSnap is for Widefield/DIC/FL
14. Open up IPLAB Software:
*You should always get an error
message. It is ok.
15. Tell software which camera you’re using:
- Select “Camera” option
- Select “Select Camera”
- Click “OK” by camera
16. Setup camera settings:
- Select “Camera” option
- Select “Get Camera Info”
- Can adjust settings
- Gain Index = 3
- Speed Table = 1
- Mult Gain = 3700
17. Adjust Magnification Dial:
- Located on bottom right of
- “1x” means no additional
18. Plug in heater controller to stage dish:
- Plug temperature probe into
small hole on side of setup.
- Attach blue clips to each side.
- Use the gold ring insert for most
MatTek dishes.
*Other ring inserts available.
Turn off Microscope
Turn lasers to zero in AOTF Controller Software.
Turn off Power to Perfect Focus box.
Flip lasers to “standby”.
Turn laser key to “off”.
*Listen for fan to get quiet before turning anything else off.
Remove sample and clean-up. Liquid waste is nearby and
should be emptied if needed.
Leave the stage dish in place.
Turn off the rest of the equipment once hear fan power
*IPLAB must be closed before camera is turned off.
Put cover on microscope.
Sign log sheet.

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