8. Bruker TruLive3D Using the photo-manipulation (PM) feature

 For questions, comments or concerns please email us at biof-imaging@colorado.edu 

The Bruker TruLive3D has a photo-manipulation (PM) feature that you can use to ablate specific areas of your sample.

Calibration of the PM module

Before performing any kind of photo-manipulation, the PM module needs to be calibrated. 

Click on the Calibration tab and open the Photomanipulation Calibration panel:


Define the PM event

The next step is to define the photomanipulation event in your sample. 

Locate the gel sample with BlackFly.png

download.png

NOTE: Before running and experiment, check if the intensity of the object imaged with the ablation dichroic is within 10% of the intensity of the object imaged with an empty dichroic.

Enable PM in direct mode.png

NOTE: We recommend that you experiment  with the photomanipulation on an area of your sample that does not present any interest before moving on to what you want to image in your experiment

Below is an example of ablation with a circular shape.

Set up an experiment with a PM event

Once you have decided on where and how to perform the photomanipulation, you need to set up the experiment.

A typical photomanipulation experiment should follow this format:

Create a Channel

Always update the channel if any change is made!

Create a z-stack

PM experiment define a stack.png

Create a PM object

Create Event 0

This is the event with the preliminary z-stack before photomanipulation.

Define the tasks:

Define a trigger:

Create Event 1

This is the event where the photomanipulation happens.

Define the tasks:

NOTE: Click on the settings icon next to the pm objects to define which z plane(s) should be photomanipulated!

Define the trigger:

NOTE: Allowing the event to start after a few seconds is a good idea.

Create Event 2

This is the settings for the imaging experiment.

Define the tasks:

Set the trigger:

Run the experiment

Always save your settings for future reference!

Look at the experiment with the image viewer in LuxBundle

Go to the plane where you performed the ablation and you should be able to see it!

PM experiment viewer 2.png

Current saving process with LuxBundle

As of November 2023, the software does not create a ims header or a big data viewer header for an experiment that contains a PM event. This is an error from LuxBundle and they are trying to fix it. Unfortunately, this bug also prevents the post-processing creation of ims headers. This means that you won't be able to look at your data in Imaris or Big Data Viewer by Fiji.

Do not create an experiment that contains a PM event! Instead:

Always reach out to the Beckman center staff if you are in doubt about the process.


Revision #7
Created 14 August 2023 15:48:28 by Evolene Premillieu
Updated 20 May 2024 18:56:24 by Evolene Premillieu