Bruker TruLive3D emission filters and fluorophores spectrum
For questions, comments or concerns please email us at biof-imaging@colorado.edu
This page will help you select which filter combination you need for most commonly used fluorophores on the Bruker TruLive3D.
The Bruker TruLive3D light sheet microscope has two imaging cameras with two motorized filter wheels and two sets of filters. It is designed to split different emission wavelengths between the two cameras for simultaneous imaging with two different emission channels.
The "Long Camera" will record the fluorescence emitting at the longest wavelength and the "Short Camera" will record the fluorescence emitting at the shortest wavelength.
When you set up your experiment you want to make sure that you select the best filter option for each camera so that the fluorescence from one fluorophore does not mix up with the other.
The Bruker TruLive3D has the following emission filters options:
Filter wheel Short Camera: BP418-462 LP498 BP497-554 BP580-627 BP610-651
Filter wheel Long Camera: LP498 BP497-554 BP580-627 BP610-651 BP655-704
Below is a list of the most commonly used fluorophores and the filters you should use:
| Fluorophores | Filter | Bruker Laser line |
| GFP | BP 497-554 | 488 |
| DAPI | BP 418-462 | 405 |
| TRITC | BP 580-627 | 561 |
| mCherry | BP 580-627 | 561 |
| AlexaFluor488 | BP 497-554 | 488 |
| AlexaFluor555 | BP 580-627 | 561 |
| AlexaFluor594 | BP 580-627 or BP 610-651 | 561 |
| AlexaFluor647 | BP 655-704 | 640 |
To select the appropriate filters you need to know the emission peak wavelength of the fluorophores you are using.
For a better understanding on how to select the correct filter, you can look at the excitation/emission spectrums from fpbase.org or aatbio.com for example. When you click on the link you can use the interactive spectrum viewer and see what the filters cover.
GFP : https://www.fpbase.org/protein/egfp/
mCherry : https://www.fpbase.org/protein/mcherry/
AlexaFluor488 : https://www.aatbio.com/fluorescence-excitation-emission-spectrum-graph-viewer/alexa_fluor_488
AlexaFluor555 : https://www.aatbio.com/fluorescence-excitation-emission-spectrum-graph-viewer/xfd555_alexa_fluor_555_equivalent
AlexaFluor594 : https://www.aatbio.com/fluorescence-excitation-emission-spectrum-graph-viewer/xfd594_alexa_fluor_594_equivalent
AlexaFluor647 : https://www.aatbio.com/fluorescence-excitation-emission-spectrum-graph-viewer/xfd647_alexa_fluor_647_equivalent
If your fluorophores are not on this page please let us know and we will update this page for you!