Level-Adjustable Sample Holder With Insert For Piezo Stage (Elyra); Stage Leveling Using X-Cite Or Laser In Fluorescence - Zeiss LSM 880 Operating Manual

ZEISS

Level-adjustable Sample Holder with Insert for Piezo Stage (ELYRA)

8 Level-adjustable Sample Holder with Insert for Piezo Stage
(ELYRA)
Caution! The adjustment screws and other part of adjustable sample holders may collide with
parts of the microscope, e.g. the condenser unit. When moving a motorized stage, avoid any
collisions. Do not install inserts while the stage is moving. Be careful when switching the
condenser front lens.
8.1

Stage Leveling Using X-Cite or Laser in Fluorescence

Adjustable sample holder – schematic
Fig. 5
view
1. Take a fluorescent sample like embedded beads and mount centrally over the objective. (Note: you
can also use your own fluorescent sample). Use WF illumination with the X-Cite or laser light and
focus manually onto the sample using the stand's focus drive (Fig. 6/A).
Set stage to zero by pressing the Set Zero button in the Stage tool. Set focus to zero by pressing
2.
the Manual button in the Focus tool.
3. Adjust leveling in X in the following way:
a) Enter in the Stage tool in the Step [µm] input box "1000". Press the upper or lower pointing
arrow of the X input box, whatever is required to step 1000 µm to the right X direction.
b) If the sample is out of focus (Fig. 6/B) refocus manually by using the stand's focus drive
(Fig. 6/C); otherwise no further leveling is necessary. Record the relative change in the focus
position by reading the value in the Z-Position input box of the Focus tool. If the focus is off
more than 10 µm it is recommended to do a pre-adjustment. To this end adjust the edges of the
clefts viewed from the right side (Fig. 5/csv) and back side (Fig. 5/cbv) to be parallel using
screws sy and sx, respectively (Fig. 5). The cleft's width should be approximately 2 mm as preset
by the factory, but can be adjusted with screw sz if needed.
c) Multiply that value with 55 and manually set the focus onto this value using the stand's focus
drive (Fig. 6/D). (Note: The factor 55 is only accurate, if the sample was positioned in the center.)
d) Use the back left screw (Fig. 5/sx) of the sample holder to readjust the focus (Fig. 6/E). Turn
clockwise if the focus position was positive, counter clockwise if it was negative. (Note that one
turn corresponds to approximately 125 µm).
18
CHAPTER 4 - ANNEX
Fig. 6
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Focus control in epi-fluorescence
LSM 880
10/2014 V_01