Iron Man User Guide v. 1.4
3
Quick Guide:
1)
Turn on components in order. Don’t go too fast – PC & monitor won’t connect.
2)
Log into Windows with IdentiKey and start Zen software.
3)
Put sample on scope coverslip down. If using oil immersion, do not switch back to air
objective.
No oil on air objectives!
4)
Load or set up appropriate Channels/Tracks for imaging your sample.
5)
Turn on necessary lasers. Do not turn on lasers you won’t be using. Argon/2 and Diode go to
Standby, then On. HeNe turn on. Argon/2: adjust % Output in Laser Properties window to
40% (do not go higher than 40%). Adjust once, then leave it alone.
6)
Adjust imaging parameters –
a.
Laser attenuation %
(3-5)
b.
Frame size
(512 x 512 or 1024x1024)
c.
Scan Speed
(~4-5 is good starting point, faster for live samples)
d.
Averaging
(1)
e.
Zoom
(1)
f.
Pinhole
(1 AU)
g.
Offset
(few to no blue understaturated pixels)
h.
Gain (Master)
(few to no red oversaturated pixels)
7)
Snap for single frame image or set up parameters for Z-series or timelapse. Start Experiment.
8)
Note the details in Information tab next to your image – these are useful to write in your
imaging notes.
9)
Save and copy your data. The LMCF is not responsible for long-term data storage. See
Appendix A for information on how to transfer data to MCDB server.
10)
Lower stage away from sample, and remove sample from stage. Properly clean oil off of any
immersion objectives you may have used with lens paper. If you are at all unsure of this
process, ask for help!
11)
Leave the microscope on a low power objective for next user.
If someone is signed up within the next two hours (Check the MCDBCal!):
12)
Switch lasers to Standby (if that option is available) or Off.
13)
Close Zen.
14)
Log use in Excel Sheet, save and close.
15)
Log off Windows.
If no one is signed up within two hours or you are the last of the day:
13)
Switch lasers to Standby, then Off.
14)
Close Zen, confirming that lasers are all Off.
15)
Log use in Excel Sheet, save and close.
16)
Shut down system: Microscope base (#4), PC (#3, Start
Shutdown), Mercury lamp (#2)
17)
WAIT FOR THE FANS ON THE LASERS TO SLOW/COOL DOWN
. This takes a couple minutes, and
the room should sound like it did when you walked in. Then flip the two switches (#1a & 1b).
