Evident Fluoview 4000 (Irchel) - 2: Multichannel + Z-stack acquisition

Ocular: adjustments for viewing via the ocular and settings for DIC.

LSM Imaging: scan field, speed, galvano vs resonant scanner, scan size

Microscope: select objectives, select Z drift compensation controls

Series: adjustments of z stacks

Observation Method: Store individual observation methods

Detector Setting: Select lasers and detectors for fluorochromes / dyes and tags and specifically adjust settings

Bright Z: depth dependent adjustment of lasers and detectors

Acquire: acquisition of z stacks, timelapse or multi area detection

Galvano (slow) vs Resonant scanner selection

OneWay vs Roundtrip (bidirectional - faster, but lower resolution and quality) selection

Scan size: digital resolution

Area Settings: scan field area and zoom, rotation

Dye and Detector Select

List of dyes to add.

TD -> Transmitted light channel

Phase: channels requiring hardware changes (e.g. filter wheels)

Selection for Averages, Accumulation or sequential settings (line, frame). In sequential scanning, groups represent the sequences

Pinhole

Groups of laser / detector settings

Adjustment of lasers and bandwith of the detector (nummerical values or by colored bands in the emission spectra that can be shifted)

Selection for timelapse and / or z stacks.

Only if time or Z is on, a timelapse or stack is later acquired.

Settings for timelapse

Settings for z stacks.

Start/End: Definition via start and end position.

Range: Definition via a range. This option must be used, if multiple positions / areas will be acquired using MATL.

Acquire>Normal

Specify where your data is stored. Select a filename.

LSM Start: starts the acquistion.

Indicators (T, Z,...) show if a timeseries, z Stack are aquired

If a series is finished, press Series Done

Select Acquire>MATL

Open the addition MATL panel via the '>' icon in the live tab

Select the tab 'MATL'

Make sure you select time and z stack configuration (series tab) for your multi area acquistion before continuing.

Place individual postions, tileset, areas as needed.

For each postion, tileset, do not forget to select the overlay checkmark for you position, tileset (only 3 channels allowed). If stitching is needed, select this checkmark too.

Positions, tilesets inherit the acquisiton / detection parameters which were active at the time of generation. Updating is possible. Using active focus control is possible by storing the active focus reference with each position or tileset. This must be done after the position or tileset is defined.

Acquire all position areas by pressing start. If necessary a loop over all position / tilesets can be defined.