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Huygens Chromatic Aberration Corrector

Correct xyz shifts, rotations and scaling differences between channels




Nearly all multi-channel images show some degree of chromatic aberration. Chromatic aberration causes focal shifts and possible scaling and rotation differences between fluorescent channels. In addition, misalignment between light paths during acquisition can result in lateral (xy-direction) shifts between channels. These aberrations clearly affect any type of multi-channel analyses like colocalization and object-based analysis and should therefore be fixed beforehand. Huygens Chromatic Aberration Corrector quickly estimates, visualizes and corrects chromatic aberrations. If you suspect any additional crosstalk/bleed-through in your multi-channel data, you may want to have a look at the Huygens Crosstalk Corrector as well.

Image description
HeLa cells stained with two different fluorophores. This image shows a shifts between the two different channels which has been corrected with the Chromatic Abberation Corrector . Raw image data was provided by Dr. Yury Belyaev, ALMF, EMBL Heidelberg, Germany


Before
Corrected


Automatic and Easy

Automated estimation and correction in two clicks.


Accurate correction

Shifts visualized for easy verification.


Custom correction

Load bead images to customize to your set-up.




Testimonials

The deconvolution and chromatic shift corrector have improved our bead representation. [..] The chromatic shift was a problem with our objective but now it seems much reduced, so we are happy.

Dr. Dominic Waithe, researcher at Wolfson Imaging Centre, University of Oxford, UK.
I have been a Huygens user for 8-9 years now and the support provided by you guys are fantastic!

Dr. Priyam Banerjee, image analysis researcher and instructor, MD Anderson Cancer Center, USA.



ChromAb U2OS

Reliable analysis

The Huygens Chromatic Aberration Corrector corrects misalignments, scaling differences and rotations between fluorescent channels in a multi-channel image. This restores the true relative positions of objects in one channel with respect to those in other channels. The image can then be used to perform multi-channel analyses such as colocalization and object-based analysis.

Image description
Chromatic aberration correction reveals colocalization between labeled Mis12 (green) and Aurora B kinase (red) in a U2OS cell. Data from Dr. Livio Kleij en Martijn Vroomans, UMC Utrecht, The Netherlands.



Use in research

Ivanova, D., Imig, C., Camacho, M. et al. CtBP1-Mediated Membrane Fission Contributes to Effective Recycling of Synaptic Vesicles.
Deconvolution and Chromatic Aberration Correction were performed with Huygens.
Cell Rep. 30, 2444–2459 (2020).

Dumbović, G., Sanjuan, X., Perucho, M. et al. Stimulated emission depletion (STED) super resolution imaging of RNA- and protein-containing domains in fixed cells.
Huygens Chromatic Aberration Corrector was used to estimate shift between channels.
Methods 187, 68-76 (2021).

For more, see Scientific Publications

Multi-channel images often suffer from crosstalk (a.k.a. bleed-through), which should be corrected before performing analyses. Huygens offers a wide range of analysis options, for example the assessment of the colocalization between different molecules.

Crosstalk Corrector Colocalization Analyzer

More information

Point Spread Function
Huygens Deconvolution
Restoration examples
Chromatic Aberration



mip_beforealignment+alignment_3.png
Recording Beads is a good method to calibrate the existing chromatic aberration. The result of the analysis and correction delivered by the Chromatic Aberration Corrector on a beads image can easily be saved to a template and applied to any image taken under similar conditions. Left: MIP rendered projection of a Huygens deconvolved 3D widefield stack of 100nm 4-channel multicoloured beads (TetraSpeck - Life Technologies). Chromatic Aberration between channels can be more clearly measured and corrected after deconvolution. Right: MIP projection of the same image after using the Huygens Chromatic Aberration Corrector. Insets show a larger view of a single XZ slice of one bead.


Not aligned
Aligned
HeLa cells stained with two different fluorophores. This image shows a shifts between the two different channels which has been corrected with the Chromatic Abberation Corrector . Raw image data was provided by Dr. Yury Belyaev, ALMF, EMBL Heidelberg, Germany

CAC WindowsRightpart
Huygens Chromatic Aberration Corrector shows a table with the values defining the shift, scaling and degrees of rotation


Nups CAC
The figure shows a multi-channel confocal (green) and STED (red) image of the nuclear pore complex. The left image shows the deconvolved and chromatic aberration corrected image, the middle shows the deconvolved image before chromatic aberration correction and the right image shows the raw data. Before chromatic aberration correction, a clear shift is visible between the confocal and the STED channel. Image courtesy of Leica Microsystems.