Adjusting Channel Presets; Multidimensional Acquisition Setup - Zeiss AXIOVERT 200 Quick Start Manual

o refocus/ re-centre
or
o choose a flourescent preset
o if there is only a faint signal switch "Auto Contrast" on
o refocus/ re-centre
o switch off "Auto Contrast"

adjusting channel presets:

o choose a preset button for your channel settings by clicking on one
o standard presets for filter cubes are
o adjust
o exposure time, binning, gain and offset (make sure "Auto Contrast" is off!!!)
o bit depth
o right click on the preset button > Overwrite

multidimensional acquisition setup:

o set multiple channels
o open "Acquisition Setup" dialogue
o tick "Change channels"
o add as many channels as you need with the "+" button
o from the appearing drop down menus choose the desired channel presets
o if you don`t want to do a stack, time series or multiple positions set timepoint to 1,
choose no focus and no xy
o set stack
o before any other adjustments: click on Z position button and set "Top" and "Bottom"
to the mechanical limits
o using defined focus positions
o using Autofocus
1 = Brightfield
2 = DAPI/ Hoechst
3 = rhodamine / TRITC/ Alexa543
4 = GFP / FITC/ Alexa488
move focus to mid focus plane
click on Z position button, choose "Set Zero"
move focus to top position
click on Z position button, choose "Set Top"
move focus to bottom position
click on Z position button, choose "Set Bottom"
in "Acquisition Setup" dialogue
•
set "Change focus using" to "Zeiss Focus Drive"
•
choose and set Z-spacing or number of slices
18 Feb 2011,<qsg-wf1> p. 4/8