Protein dynamics and Deep tissue imaging
Group leader: Prof. Dr Marloes Groot

Marloes Groot


To understand cellular interactions in live tissue and develop novel diagnostic tools we use the physics of the interaction of short laser pulses with biological tissue to generate label-free microscopic images of volumes of tissue. We study the microscopic composition of skin, lung and brain, and work towards clinical applications to detect tumor cells and diagnose diseases based on label-free third harmonic generation microscopy. See here for a few examples of our images of skin and brain. In a related project we develop automated image analysis tools to quantitate the information of the volumes imaged by THG microscopy.

The movie
For a short movie of our research made end of 2020, watch hier our lab tour.


- First THG/SHG measurements on Covid-19 lung samples, measured by Laura in the Amsterdam UMC. Curious to see the correlation with histology. 

Covid-19-lung-samples  naast-covid

- The 3D skin stretcher works, plus 2D orientation analysis works! Great work by Ludo! 3D analysis by Amy in preparation. Watch the clip

- Laura van Huizen and Ludo van Haasterecht won 1st and 2nd prize at the LaserLabB Best Article 2020 competition! Congrats to both!

- The study of Enis Arik and Pat Konold got published in Nature Communications! Here we settle a longstanding dispute in literature between the x-ray diffraction and spectroscopy communities. Is the chemistry of an internal cofactor in a protein different when that protein is in a crystal or in solution? We studied the light-induced structural changes in hydrogen bond length and larger scale conformational changes, with femtosecond multi-pulse absorption spectroscopy in the midinfrared and visible regions, and achieved sub-Å resolution and 100 femtosecond time resolution on a time range of 1 millisecond, to see that the entire photocycle pathway is different.


- Laura’s article on long tumor tissue imaged with the portable THG/SHG analyzer got published in Translational Biophotonics, see Compact portable multiphoton microscopy reveals histopathological hallmarks of unprocessed lung tumor tissue in real time Translational Biophotonics. 


- Our first day with the Femtodiagnostics BV portable THG/SHG analyzer in the endoscopy suite of prof. dr Jouke Annema (Amsterdam UMC). 


- The work of Andy Zhang, with Pieter Wesseling and Philip de Witt Hamer and others got published in Advanced Science. A great achievement by Andy Zhang, who measured THG brain tumor images, developed automatic image analysis algorithms, and got neuropathologists to diagnose the images. See Advanced Science6, doi:10.1002/advs.201900163 (2019) 

AdvancedScience2019   AdvancedScience6

Read more

Our main research line is aimed at biomedical imaging in deep tissue using label free microscopy. We use the physics of the interaction of short laser pulses with biological tissue to generate label-free microscopic images of volumes of tissue, in particular through second and third harmonic generation processes. With this we pursue several lines of interest, aimed at clinical diagnostics and at the study of fundamental processes in live tissue. A short summary, citing a few highlights: We started imaging mouse brains to demonstrate the versatility of the technique1. We have used SHG/THG to detect healthy and diseased states in excised tissue2-6 and have developed automated image analysis tools for computer aided diagnosis based on classical algorithms7-9 and neural networks. Applications include label-free analysis of clinical tumor samples2-6, pathological states of connective tissue in inflammation and mechanisms underlying wound healing10-12 as well as scoring and detection of resection margin of cancer. In glioma samples, normal brain was discriminated from infiltrative glioma with 96.6% sensitivity and 95.5% specificity, in nearly perfect (93%) agreement with pathologic scoring2 and concurring results were reached in healthy breast tissue and in lung tumor tissue3,6.
We are currently bringing transportable SHG/THG tissue analysers developed by Femtodiagnotics BV (Disclosure: MLG has indirect interests in Femtodiagnostics BV) into the clinic6 and are developing THG/SHG fiberscopes to be used in-situ, to help determine what tissue to excise during surgery. Furthermore, we are developing machine learning algorithms for automated image analysis of the 3D THG/SHG images. Supported by the Dutch Burn Foundation and TKI Holland Health we study the relationship between the 3D microstructure of skin and its macroscopic properties elasticity and strength, in healthy and scar tissue.

References (For a full publication list of Marloes Groot, see CV).
1    Witte, S., Negrean, A., Lodder, J. C., de Kock, C. P. J., Silva, G. T., Mansvelder, H. D. & Groot, M. L. Label-free live brain imaging and targeted patching with third-harmonic generation microscopy. P Natl Acad Sci USA  108, 5970-5975 (2011).
2    Zhang, Z. Q., de Munck, J. C., Verburg, N., Rozemuller, A. J., Vreuls, W., Cakmak, P., van Huizen, L. M. G., Idema, S., Aronica, E., Hamer, P. C. D., Wesseling, P. & Groot, M. L. Quantitative Third Harmonic Generation Microscopy for Assessment of Glioma in Human Brain Tissue. Advanced Science6, doi:10.1002/advs.201900163 (2019).
3    van Huizen, L. M. G., Kuzmin, N. V., Barbe, E., van der Velde, S., te Velde, E. A. & Groot, M. L. Second and third harmonic generation microscopy visualizes key structural components in fresh unprocessed healthy human breast tissue. Journal of Biophotonics  12, doi:10.1002/jbio.201800297 (2019).
4    Kuzmin, N. V., Idema, S., Aronica, E., Hamer, P. C. D. W., Wesseling, P. & Groot, M. L.    (ed S. Shoham F. S. Pavone)  (CRC Press, 2019).
5    Kuzmin, N. V., Wesseling, P., Hamer, P. C., Noske, D. P., Galgano, G. D., Mansvelder, H. D., Baayen, J. C. & Groot, M. L. Third harmonic generation imaging for fast, label-free pathology of human brain tumors. Biomed Opt Express  7, 1889-1904, doi:10.1364/BOE.7.001889 (2016).
6    van Huizen, L. M. G., Radonic, T., van Mourik, F., Seinstra, D., Dickhoff, C., Daniels, J. M. A., Bahce, I., Annema, J. T. & Groot, M. L. Compact portable multiphoton microscopy reveals histopathological hallmarks of unprocessed lung tumor tissue in real time. Translational Biophotonics  n/a, e202000009, doi:10.1002/tbio.202000009 (2020).
7    Zhang, Z. Q., Groot, M. L. & de Munck, J. C. Tensor regularized total variation for denoising of third harmonic generation images of brain tumors. Journal of Biophotonics  12, doi:10.1002/jbio.201800129 (2019).
8    Zhang, Z. Q., Kuzmin, N. V., Groot, M. L. & de Munck, J. C. Quantitative comparison of 3D third harmonic generation and fluorescence microscopy images. Journal of Biophotonics  11, doi:UNSP e201600256 10.1002/jbio.201600256 (2018).
9    Zhang, Z. Q., Kuzmin, N. V., Groot, M. L. & de Munck, J. C. Extracting morphologies from third harmonic generation images of structurally normal human brain tissue. Bioinformatics  33, 1712-1720, doi:10.1093/bioinformatics/btx035 (2017).
10    Bos, E. J., Pluemeekers, M., Helder, M., Kuzmin, N., van der Laan, K., Groot, M. L., van Osch, G. & van Zuijlen, P. Structural and Mechanical Comparison of Human Ear, Alar, and Septal Cartilage. Prs-Glob Open  6, doi:ARTN e1610 10.1097/GOX.0000000000001610 (2018).
11    Jaspers, M. E. H., Brouwer, K. M., van Trier, A. J. M., Groot, M. L., Middelkoop, E. & van Zuijlen, P. P. M. Effectiveness of Autologous Fat Grafting in Adherent Scars: Results Obtained by a Comprehensive Scar Evaluation Protocol. Plast Reconstr Surg  139, 212-219, doi:10.1097/Prs.0000000000002891 (2017).
12    Visscher, D. O., Bos, E. J., Peeters, M., Kuzmin, N. V., Groot, M. L., Helder, M. N. & van Zuijlen, P. P. M. Cartilage Tissue Engineering: Preventing Tissue Scaffold Contraction Using a 3D-Printed Polymeric Cage. Tissue Eng Part C-Me  22, 573-584, doi:10.1089/ten.tec.2016.0073 (2016).