Invitrogen EVOS M5000 - Page 66

5.
(Optional) If the EVOS
™
Onstage Incubator is installed and used for a time-lapse routine, click Use
Incubator.
Note: This option is only available if you have installed and connected the EVOS
™
Onstage
Incubator to your EVOS
™
M5000instrument.
IMPORTANT! Before using the EVOS
™
Onstage Incubator in your time-lapse experiments, ensure
that:
·
The EVOS
™
Onstage Incubator has been set up for use.
·
The gas inputs have been configured.
·
The oxygen sensor has been calibrated.
6.
Adjust brightness and focus for each selected channel as described in “Capture images in a single
channel”
on page28, then click Next.
7.
If you have opted to run
Auto Focus, select the Auto Focus channel (for example, DAPI, GFP, and
so on) to use.
Note:
In a time lapse routine, you can run the autofocus procedure only in a single channel. The
focal plane
identified in this channel is then used for all other channels.
8.
Select to autofocus at
Every interval or at First interval of each run, then click Next.
Note:
In a time-lapse routine, images are captured at the end of each interval of a run. For more
information, see “Intervals”
on page68.
9.
If you have opted to capture
Z-Stack images, define the Top, In Focus, and Bottom positions for
the Z-Stack image set.
a.
If not already on, click the Light button to illuminate the sample as you locate the boundaries
and the In Focus position of the Z-Stack.
b.
To accept the
Current Z as the In Focus position, click Set.
To change the In Focus position, move the Coarse and Fine focus sliders on the Capture
tab to focus on the sample, then click Set.
c.
Move the focus sliders up from the Current Z position to the desired Top position (upper
boundar
y of the Z-Stack), then click Set for Top.
d.
Move the focus sliders down from the Current Z position to the desired Bottom position
(lower boundar
y of the Z-Stack), then click Set for Bottom.
Note:
Click
Z number for the Top, In Focus, and Bottom positions to jump to that position along
the Z-axis.
Chapter7Capture time lapse images
Run a time lapse routine
7
66
EVOS
™
M5000 Imaging System User Guide