Live tracking of moving samples in confocal microscopy for vertically grown roots

Area of Science:

• Plant Biology
• Developmental Biology
• Microscopy and Imaging

Background:

• Root growth involves integrating environmental signals for directional orientation.
• Studying root development in *Arabidopsis* is crucial but challenging due to imaging limitations.
• Live imaging of dynamic biological processes requires advanced tracking techniques.

Purpose of the Study:

• To develop an improved confocal microscopy setup for vertical sample mounting.
• To create custom software (TipTracker) for automatic tracking of moving biological objects.
• To enable long-term observation of root tip growth and response to stimuli.

Main Methods:

• A novel confocal microscope setup with vertical sample mounting and directional illumination was engineered.
• Custom software, TipTracker, was developed for automated object tracking in microscopy.
• The system was tested on *Arabidopsis* root tips and migrating zebrafish embryo cells.

Main Results:

• The setup allows for prolonged observation of root tips growing along the gravity vector.
• Rapid gravity and light stimulation experiments were successfully performed.
• High-resolution data sets of dynamic samples, including migrating cells, were acquired.

Conclusions:

• The combined microscopy setup and TipTracker software facilitate high-resolution live imaging of dynamic biological samples.
• This system overcomes previous limitations in observing root growth and cell migration.
• The tools are adaptable for implementation on other microscopy platforms.