Stimulated Raman scattering microscopy in the analysis of cancer cells – A review and own research
TrAC Trends in Analytical Chemistry, 2023, 169, IF=13.1, MEnS=200
A. Adamczyk1, S. Orzechowska1, A. Nowakowska1, K. Brzozowski1, K. Majzner, M1. Baranska1,2
1Faculty of Chemistry, Jagiellonian University, Gronostajowa 2, 30-387, Krakow, Poland
2Jagiellonian Centre for Experimental Therapeutics (JCET), Jagiellonian University, Bobrzynskiego 14, 30-348, Krakow, Poland
Abstract
This article shows the latest advances in the spectroscopic analysis of cancer cells to identify pathological changes and understand the mechanism of metastasis using stimulated Raman Scattering (SRS) microscopy. SRS is a nonlinear technique that probes vibrational modes of molecules and offers several advantages over conventional Raman techniques, including much stronger signals and the ability to time-resolve the vibrational motions. Consequently, SRS microscopy offers analysis of single cells in a rapid way. On the one hand, the development of modern Raman imaging technology based on nonlinear optics is shown, and on the other hand, the latest achievements of this technology in the analysis of various tumours cells, in particular leukaemia. Additionally, the results of own research performed on the home-built SRS setup are presented and the perspectives of analysis based on SRS. Considering the advantages of SRS microscopy, it is expected that it will support cancer diagnostics in clinical practice.
Acknowledgments
This work was supported by the “Label-free and rapid optical imaging, detection and sorting of leukaemia cells” project and is carried out within the Team-Net program (POIR.04.04.00-00-16ED/18-00) of the Foundation for Polish Science co-financed by the European Union under the European Regional Development Fund. This work was partially supported by the "Excellence Initiative - Research University" program at the Jagiellonian University in Krakow ("Young Laboratories" Program (Edition 2), “Real-time analysis of metabolism of live cancer cells by means of stimulated Raman spectroscopy”, U1U/W20/NO/17).
The authors are grateful to MSc Adrianna Wislocka-Orlowska for help with sample preparation.
Automatic subtyping of Diffuse Large B-cell Lymphomas (DLBCL): Raman-based genetic and metabolic classification
Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 2023 (accepted), IF=4.4, MEnS=140
Patrycja Dawiec1,2, Patrycja Leszczenko1,2, Anna M. Nowakowska1, Paulina Laskowska3, Maciej Szydłowski3, Przemysław Juszczyński3, Malgorzata Barańska1,4, Piotr Mrówka3,5*, Katarzyna Majzner1*
1Department of Chemical Physics, Faculty of Chemistry, Jagiellonian University, Gronostajowa 2, 30-387 Krakow, Poland
2Doctoral School of Exact and Natural Sciences, Jagiellonian University, Łojasiewicza 11, 30-348 Krakow, Poland
3Department of Experimental Hematology, Institute of Hematology and Transfusion Medicine, Warsaw Poland
4Jagiellonian Centre for Experimental Therapeutics, Jagiellonian University, Bobrzyńskiego 14, 30-348 Krakow, Poland
5Department of Biophysics, Physiology and Pathophysiology, Medical University of Warsaw, Poland
Reliable Cell Preparation Protocol for Raman Imaging to Effectively Differentiate Normal Leukocytes and Leukemic Blasts
Spectrochimica Acta Part A: Molecular and Biomolecular Spectroscopy, 2023, 292, 1-13, 122408, IF=4.831, MEnS=140
A. M. Nowakowska1, A. Borek-Dorosz1, P. Leszczenko1,5, A. Adamczyk1,5, A. Pieczara2,5, J. Jakubowska3, A. Pastorczak3, K. Ostrowska3, K. M. Marzec4, K. Majzner1,2*
1Jagiellonian University in Kraków, Faculty of Chemistry, Department of Chemical Physics, 2 Gronostajowa St., Krakow, Poland
2Jagiellonian University in Kraków, Jagiellonian Centre for Experimental Therapeutics (JCET), 14 Bobrzynskiego St., Krakow, Poland
3Department of Pediatrics, Oncology and Hematology, Medical University of Lodz, Sporna St. 36/50, Łodz, Poland
4Lukasiewicz Research Network - Krakow Institute of Technology, 73 Zakopianska St., Krakow, Poland
5Jagiellonian University in Kraków, Doctoral School of Exact and Natural Sciences, 11 Lojasiewicza St., Krakow, Poland
Abstract
Leukemias are a remarkably diverse group of malignancies originating from abnormal progenitor cells in the bone marrow. Leukemia subtypes are classified according to the cell type that has undergone neoplastic transformation using demanding and time-consuming methods. Alternative is Raman imaging that can be used both for living and fixed cells. However, considering the diversity of leukemic cell types and normal leukocytes, and the availability of different sample preparation protocols, the main objective of this work was to verify them for leukemia and normal blood cell samples for Raman imaging.
The effect of glutaraldehyde (GA) fixation in a concentration gradient (0.1 %, 0.5 %, and 2.5 % GA) on the molecular structure of T-cell acute lymphoblastic leukemia (T-ALL) and peripheral blood mononuclear cells (PBMCs) was verified. Changes in the secondary structure of proteins within cells were indicated as the main effect of fixation, as shown by an increase in band intensity at 1041 cm−1, characteristic for in-plane δ(Csingle bondH) deformation in phenylalanine (Phe).
Different sensitivity of mononuclear and leukemic cells to fixation was observed. While the 0.1 % concentration of GA was too low to preserve the cell structure for an extended period of time, a GA concentration of 0.5 % seemed optimal for both normal and malignant cells. Chemical changes in PBMCs samples stored for 11 days were also investigated, which manifested in numerous modifications in the secondary structure of proteins and the content of nucleic acids. The impact of cell preculturing for 72 h after unbanking was verified, and there was no significant effect on the molecular structure of cells fixed with 0.5 % GA. In summary, the developed protocol for the preparation of samples for Raman imaging allows for the effective differentiation of fixed normal leukocytes from malignant T lymphoblasts.
Acknowledgements
This work was supported by the “Label-free and rapid optical imaging, detection and sorting of leukemia cells” project and is carried out within the Team-Net program (POIR.04.04.00–00-16ED/18–00) of the Foundation for Polish Science co-financed by the European Union under the European Regional Development Fund.
Authors would like to address special gratitude to Professor Malgorzata Baranska for her invaluable support and Ms. Adrianna Wisłocka-Orłowska, Ms. Klaudia Mielnik and Dr. Krzysztof Brzozowski (Jagiellonian University, Faculty of Chemistry, Krakow, Poland) for their kind assistance in data acquisition.
więcej oRaman Spectroscopy Imaging As a Tool to Classify Diffused Large B-Cell Lymphoma Subtypes
Abstract, Blood, 142, 1, 2023, 5040
P.Dawiec, P. Laskowska, P. Leszczenko, A. M. Nowakowska, M. Szydlowski, P. Juszczyński, M. Baranska, K. Majzner, P. Mrówka
więcej oLabel-Free Identification of Biochemical Footprints of Key Mutations in Acute Myeloid Leukemia Using Raman Spectroscopy
Abstract, Blood, 142, 1, 2023, 6022
P. Laskowska, S. Orzechowska, A. Borek-Dorosz, A. M. Nowakowska, W. Korona, A. Adamczyk, P. Leszczenko, A. Szlachetka, M. Zasowska, M. Szydlowski, P. Juszczyński , M. Baranska, K.Majzner, P. Mrówka
więcej oRaman Spectroscopy Imaging in Evaluation of Immune Cells Activation Status
Abstract, Blood, 142, 1, 2023, 7157
P. Laskowska, A. Borek-Dorosz, A. M. Nowakowska, A. Zolyniak, E. Kurtz, M. Szydlowski, P. Juszczyński, M. Baranska, K. Majzner, P. Mrówka
więcej o