RCF Microscopy Manual
5/2/2017
Authors Cory Nook and Chaowei Shang
Page 10 of 19
4.2 Aptome Settings
About the Aptome
The ApoTome uses a projected grid along with a
mathematical algorithm to remove out-of-focus
light, similar to the function of the pinhole on a
confocal microscope. When not in use, it should be
pulled out slightly from the microscope. There are
three small grids (L, H, D) that can be used in the
ApoTome; each objective requires a particular grid.
The software will prompt you to change the grid if it
senses that the grid and the chosen objective do not
match. Sometimes, because objectives are retired
and new ones created, the match between
objective and grid may not be reflected in the
software. A Zeiss engineer helped me come up with
the best pairing for objectives and grids that we
have (see below).
When the ApoTome is pushed in all the way, it is
engaged. The grid moves through 3 positions as it
captures one image. When you need to disengage
the ApoTome from the microscope, turn off its
power supply first (Labeled #4), which allows the
grid to return to its original position,
then turn in
back ON
before you pull it part or all of the way out.
Failure to do so may cause damage to both the grid
and the Apotome. Should the grid become loose
from its niche in the Apotome, let me know, and I
will retrieve it.
From the menu, choose the ApoTome button. A dialogue box
labelled ApoTome will pop up. Choose the second tab, Settings,
and choose the following:
ApoTome: Live Mode – Grid visible
ApoTome: Acquisition Mode – Optional Sectioning
ApoTome Grid – Choose the correct grid for your
objective from the drop down menu
ApoTome Filter – Medium
ApoTome: Averaging (Noise Reduction) – 2
ApoTome: Status – this should say Calibrated in a green
field (if not, come get me!)
Obj
Objective
Medium Grid
10x EC Plan-Neofluar 10x/0.3 DIC I
Air
L
20x Plan-Apochromat 20x/0.8 DIC II
Air
L
40x LD Plan-Neofluar 40x/0.6 Ph2 DIC II
Air
NA*
40x Plan-Apochromat 40x/1.3 DIC (UV) VIR-IR
Oil
L
63x Plan-Apochromat 63x/1.4 DIC III
Oil
H
100x Plan-Apochromat 100x/1.4
Oil
D
*The 40X Air Objective is a Phase Objective and shouldn’t be used with the Apotome.
