{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,7,7]],"date-time":"2026-07-07T17:00:28Z","timestamp":1783443628176,"version":"3.54.6"},"reference-count":63,"publisher":"MDPI AG","issue":"19","license":[{"start":{"date-parts":[[2020,9,27]],"date-time":"2020-09-27T00:00:00Z","timestamp":1601164800000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Italian Ministry for University and Research (MIUR)","award":["PON NeON\/ARS01_00769"],"award-info":[{"award-number":["PON NeON\/ARS01_00769"]}]},{"DOI":"10.13039\/501100005010","name":"Associazione Italiana per la Ricerca sul Cancro","doi-asserted-by":"publisher","award":["21420"],"award-info":[{"award-number":["21420"]}],"id":[{"id":"10.13039\/501100005010","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Sensors"],"abstract":"<jats:p>In the last decade, Raman Spectroscopy (RS) was demonstrated to be a label-free, non-invasive and non-destructive optical spectroscopy allowing the improvement in diagnostic accuracy in cancer and analytical assessment for cell sensing. This review discusses how Raman spectra can lead to a deeper molecular understanding of the biochemical changes in cancer cells in comparison to non-cancer cells, analyzing two key examples, leukemia and breast cancer. The reported Raman results provide information on cancer progression and allow the identification, classification, and follow-up after chemotherapy treatments of the cancer cells from the liquid biopsy. The key obstacles for RS applications in cancer cell diagnosis, including quality, objectivity, number of cells and velocity of the analysis, are considered. The use of multivariant analysis, such as principal component analysis (PCA) and linear discriminate analysis (LDA), for an automatic and objective assessment without any specialized knowledge of spectroscopy is presented. Raman imaging for cancer cell mapping is shown and its advantages for routine clinical pathology practice and live cell imaging, compared to single-point spectral analysis, are debated. Additionally, the combination of RS with microfluidic devices and high-throughput screening for improving the velocity and the number of cells analyzed are also discussed. Finally, the combination of the Raman microscopy (RM) with other imaging modalities, for complete visualization and characterization of the cells, is described.<\/jats:p>","DOI":"10.3390\/s20195525","type":"journal-article","created":{"date-parts":[[2020,9,28]],"date-time":"2020-09-28T08:02:58Z","timestamp":1601280178000},"page":"5525","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":49,"title":["Raman Microscopy: Progress in Research on Cancer Cell Sensing"],"prefix":"10.3390","volume":"20","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-6309-7880","authenticated-orcid":false,"given":"Satheeshkumar","family":"Elumalai","sequence":"first","affiliation":[{"name":"Institute of Biochemistry and Cell Biology (IBBC), National Research Council of Italy (CNR), Via P. Castellino 111, 80131 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7445-385X","authenticated-orcid":false,"given":"Stefano","family":"Manag\u00f3","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry and Cell Biology (IBBC), National Research Council of Italy (CNR), Via P. Castellino 111, 80131 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-3696-8465","authenticated-orcid":false,"given":"Anna Chiara","family":"De Luca","sequence":"additional","affiliation":[{"name":"Institute of Biochemistry and Cell Biology (IBBC), National Research Council of Italy (CNR), Via P. Castellino 111, 80131 Naples, Italy"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2020,9,27]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"501","DOI":"10.1038\/121501c0","article-title":"A new type of secondary radiation","volume":"121","author":"Raman","year":"1928","journal-title":"Nature"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"477","DOI":"10.1038\/122477a0","article-title":"The Raman Effect in Crystals","volume":"122","author":"Krishnan","year":"1928","journal-title":"Nature"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"203","DOI":"10.1098\/rspa.1953.0057","article-title":"Intensities in Raman spectra I. A bond polarizability theory","volume":"217","author":"Long","year":"1953","journal-title":"Proc. R. Soc. Lond. A"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"7","DOI":"10.1016\/j.optlastec.2018.06.034","article-title":"Raman microscopy-based sensing of leukemia cells: A review","volume":"108","author":"Zito","year":"2018","journal-title":"Opt. Laser Technol."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"5766","DOI":"10.1021\/cr300147r","article-title":"Raman imaging in biochemical and biomedical applications: Diagnosis and treatment of breast cancer","volume":"113","author":"Abramczyk","year":"2013","journal-title":"Chem. Rev."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"25888","DOI":"10.1039\/C8RA04491K","article-title":"Challenges in application of Raman spectroscopy to biology and materials","volume":"8","author":"Kuhar","year":"2018","journal-title":"RSC Adv."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"141","DOI":"10.3390\/bios5020141","article-title":"Label-free imaging and biochemical characterization of bovine sperm cells","volume":"5","author":"Ferrara","year":"2015","journal-title":"Biosensors"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"24821","DOI":"10.1038\/srep24821","article-title":"A reliable Raman-spectroscopy based approach for diagnosis, classification and follow-up of B-cell acute lymphoblastic leukemia","volume":"6","author":"Valente","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_9","doi-asserted-by":"crossref","unstructured":"Chaturvedi, D., Balaji, S.A., Bn, V.K., Ariese, F., Umapathy, S., and Rangarajan, A. (2016). Different phases of breast cancer cells: Raman study of immortalized, transformed, and invasive cells. Biosensors, 6.","DOI":"10.3390\/bios6040057"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"2070","DOI":"10.1039\/C9AN01811E","article-title":"Applications of Raman spectroscopy in the development of cell therapies: State of the art and future perspectives","volume":"145","author":"Rangan","year":"2020","journal-title":"Analyst"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"99","DOI":"10.1080\/14737159.2020.1724092","article-title":"A look into the use of Raman spectroscopy for brain and breast cancer diagnostics: Linear and non-linear optics in cancer research as a gateway to tumor cell identity","volume":"20","author":"Abramczyk","year":"2020","journal-title":"Expert Rev. Mol. Diagn."},{"key":"ref_12","doi-asserted-by":"crossref","unstructured":"Pal, U.M., Saxena, M., Anil Vishnu, G.K., Parsana, D., Sarvani, B.S.R., Varma, M., Jayachandra, M., Kurpad, V., Baruah, D., and Gogoi, G. (2020). Optical spectroscopy-based imaging techniques for the diagnosis of breast cancer: A novel approach. Appl. Spectrosc. Rev., 1\u201327.","DOI":"10.1080\/05704928.2020.1749651"},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Matthews, R., and Choi, M. (2016). Clinical utility of positron emission tomography magnetic resonance imaging (PET-MRI) in gastrointestinal cancers. Diagnostics, 6.","DOI":"10.3390\/diagnostics6030035"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"11","DOI":"10.1038\/s41698-020-0114-1","article-title":"Multiplex bioimaging of single-cell spatial profiles for precision cancer diagnostics and therapeutics","volume":"4","author":"Allam","year":"2020","journal-title":"NPJ Precis. Onc."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"247","DOI":"10.1016\/j.infrared.2018.08.005","article-title":"Histochemical analysis of human breast tissue samples by IR and Raman spectroscopies. Protocols discussion","volume":"93","author":"Kopec","year":"2018","journal-title":"Infrared Phys. Technol."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"275","DOI":"10.1016\/S0091-679X(08)00610-9","article-title":"Infrared and Raman microscopy in cell biology","volume":"89","author":"Bird","year":"2008","journal-title":"Methods Cell Biol."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"691","DOI":"10.1007\/s10555-018-9770-9","article-title":"Applications of Raman spectroscopy in cancer diagnosis","volume":"37","author":"Auner","year":"2018","journal-title":"Cancer Metastasis Rev."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"303","DOI":"10.1007\/s10549-014-3182-5","article-title":"Confocal fluorescence microscopy for rapid evaluation of invasive tumor cellularity of inflammatory breast carcinoma core needle biopsies","volume":"149","author":"Dobbs","year":"2015","journal-title":"Breast Cancer Res. Treat."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1155","DOI":"10.1039\/c3an01993d","article-title":"Label-free imaging of drug distribution and metabolism in colon cancer cells by Raman microscopy","volume":"139","author":"Petersen","year":"2014","journal-title":"Analyst"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"12408","DOI":"10.1073\/pnas.1301379110","article-title":"A small animal Raman instrument for rapid, wide-area, spectroscopic imaging","volume":"110","author":"Bohndiek","year":"2013","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"5844","DOI":"10.1073\/pnas.0710575105","article-title":"Noninvasive molecular imaging of small living subjects using Raman spectroscopy","volume":"105","author":"Keren","year":"2008","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"738","DOI":"10.1021\/ac9026737","article-title":"Online fluorescence suppression in modulated Raman spectroscopy","volume":"82","author":"Mazilu","year":"2010","journal-title":"Anal. Chem."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"933","DOI":"10.1364\/OE.19.000933","article-title":"Optical eigenmodes; exploting the quadratic nature of the energy flux and of scattering interactions","volume":"19","author":"Mazilu","year":"2011","journal-title":"Opt. Exp."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"037002","DOI":"10.1117\/1.3556722","article-title":"Modulated Raman spectroscopy for enhanced identification of blabber tumor cells in urine samples","volume":"16","author":"Canetta","year":"2011","journal-title":"J. Biomed. Opt."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Rehman, I.U., Movasaghi, Z., and Rehman, S. (2012). Vibrational Spectroscopy for Tissue Analysis, CRC Press, Taylor & Francis Group.","DOI":"10.1201\/b12949"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"6512","DOI":"10.1038\/s41598-017-06936-z","article-title":"Molecular profiling of single organelles for quantitative analysis of cellular heterogeneity","volume":"7","author":"Kuzmin","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"10985","DOI":"10.1021\/acs.analchem.7b02822","article-title":"Macromolecular profiling of organelles in normal diploid and cancer cells","volume":"89","author":"Levchenko","year":"2017","journal-title":"Anal. Chem."},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Kuzmin, A.N., Pliss, A., Rzhevskii, A., Lita, A., and Larion, M. (2018). BCAbox algorithm expands capabilities of Raman microscope for single organelles assessment. Biosensors, 8.","DOI":"10.3390\/bios8040106"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"6387","DOI":"10.1039\/C6AN01018K","article-title":"Rapid acquisition of mean Raman spectra of eukaryotic cells for a robust single cell classification","volume":"141","author":"Schie","year":"2016","journal-title":"Analyst"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"S35","DOI":"10.1016\/j.ijantimicag.2015.10.014","article-title":"Raman Spectroscopy towards clinical application: Drug monitoring and pathogen identification","volume":"46","author":"Neugebauer","year":"2015","journal-title":"Int. J. Antimicrob. Agents"},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"e201700265","DOI":"10.1002\/jbio.201700265","article-title":"Raman detection and identification of normal and leukemic hematopoietic cells","volume":"11","author":"Mirabelli","year":"2018","journal-title":"J. Biophotonics"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1111\/j.1749-0774.2010.00087.x","article-title":"History of leukemia-lymphoma cell lines","volume":"23","author":"Drexler","year":"2010","journal-title":"Hum. Cell"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"685","DOI":"10.1016\/0145-2126(82)90085-6","article-title":"Establishment and characterization of a human ebv-negative b cell line (MN 60)","volume":"6","author":"Roos","year":"1982","journal-title":"Leuk. Res."},{"key":"ref_34","first-page":"1783","article-title":"Proposals for the immunological classification of acute leukemias. European Group for the Immunological Characterization of Leukemias (EGIL)","volume":"9","author":"Bene","year":"1995","journal-title":"Leukemia"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"127","DOI":"10.1038\/s12276-018-0150-x","article-title":"The Implications of signaling lipids in cancer metastasis","volume":"50","author":"Luo","year":"2018","journal-title":"Exp. Mol. Med."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1097\/GCO.0b013e3283414e87","article-title":"HER2-positive breast cancer: Current management of early, advanced, and recurrent disease","volume":"23","author":"Callahan","year":"2011","journal-title":"Curr. Opin. Obstet. Gynecol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"12371","DOI":"10.1073\/pnas.0501390102","article-title":"Diagnosing breast cancer by using Raman spectroscopy","volume":"102","author":"Haka","year":"2005","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"3220","DOI":"10.1039\/c0an00524j","article-title":"A comparison of breast cancer tumor cells with varying expression of the Her2\/neu receptor by Raman microspectroscopic imaging","volume":"135","author":"Hartsuiker","year":"2010","journal-title":"Analyst"},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"025001","DOI":"10.1117\/1.JBO.19.2.025001","article-title":"Evaluating HER2 amplification status and acquired drug resistance in breast cancer cells using Raman spectroscopy","volume":"19","author":"Bi","year":"2014","journal-title":"J. Biomed. Opt."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"5646","DOI":"10.1039\/C6AN00859C","article-title":"Epigenetic changes in cancer by Raman imaging, fluorescence imaging, AFM and scanning near-field optical microscopy (SNOM). Acetylation in normal and human cancer breast cells MCF10A, MCF7 and MDA-MB-231","volume":"141","author":"Abramczyk","year":"2016","journal-title":"Analyst"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"461","DOI":"10.1177\/1533034616655953","article-title":"Label-free Raman imaging to monitor breast tumor signatures","volume":"16","author":"Manciu","year":"2016","journal-title":"Technol. Cancer Res. Treat."},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"e201700207","DOI":"10.1002\/jbio.201700207","article-title":"Internalization kinetics and cytoplasmic localization of functionalized diatomite nanoparticles in cancer cells by Raman imaging","volume":"11","author":"Migliaccio","year":"2018","journal-title":"J. Biophotonics"},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"727","DOI":"10.1177\/0003702819834575","article-title":"Applications of spatial light modulators in Raman spectroscopy","volume":"73","author":"Sinjab","year":"2019","journal-title":"Appl. Spectrosc."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"2993","DOI":"10.1364\/BOE.7.002993","article-title":"Tissue diagnosis using power-sharing multifocal Raman micro-spectroscopy and auto-fluorescence imaging","volume":"7","author":"Sinjab","year":"2016","journal-title":"Biomed. Opt. Exp."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"257","DOI":"10.1364\/OPTICA.1.000257","article-title":"Nonredundant Raman imaging using optical eigenmodes","volume":"1","author":"Kosmeier","year":"2014","journal-title":"Optica"},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Guerrini, L., and Alvarez-Puebla, R.A. (2019). Surface-enhanced Raman spectroscopy in cancer diagnosis, prognosis and monitoring. Cancers, 11.","DOI":"10.3390\/cancers11060748"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"5612","DOI":"10.1002\/smll.201601829","article-title":"Multiplexed molecular imaging of fresh tissue surfaces enabled by convection-enhanced topical staining with SERS-coded nanoparticles","volume":"12","author":"Wang","year":"2016","journal-title":"Small"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"25","DOI":"10.1016\/j.aca.2012.11.007","article-title":"Sample size planning for classification models","volume":"760","author":"Beleites","year":"2013","journal-title":"Anal. Chim. Acta"},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"2575","DOI":"10.1021\/nn406200y","article-title":"Reproducible surface-enhanced Raman quantification of biomarkers in multicomponent mixtures","volume":"8","author":"Mazilu","year":"2014","journal-title":"ACS Nano"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"12406","DOI":"10.1021\/acsami.7b19285","article-title":"Bioderived three-dimensional hierarchical nanostructures as efficient surface-enhanced Raman scattering substrates for cell membrane probing","volume":"10","author":"Zito","year":"2018","journal-title":"ACS Appl. Mater Interfaces"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"6163","DOI":"10.1039\/C5AN01074H","article-title":"Towards high-throughput microfluidic Raman-activated cell sorting","volume":"140","author":"Zhang","year":"2015","journal-title":"Analyst"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"2023","DOI":"10.1021\/acs.analchem.7b04127","article-title":"High-throughput screening Raman spectroscopy platform for label-free cellomics","volume":"90","author":"Schie","year":"2018","journal-title":"Anal. Chem."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"12653","DOI":"10.1038\/s41598-019-48895-7","article-title":"New perspectives for viability studies with high-content analysis Raman spectroscopy (HCA-RS)","volume":"9","author":"Mondol","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"L17","DOI":"10.1016\/j.bpj.2014.01.002","article-title":"Simultaneous Raman microspectroscopy and fluorescence imaging of bone mineralization in living zebrafish larvae","volume":"106","author":"Bennet","year":"2014","journal-title":"Biophy. J."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"1135","DOI":"10.1364\/OL.33.001135","article-title":"Combined Raman spectroscopy and optical coherence tomography device for tissue characterization","volume":"33","author":"Patil","year":"2008","journal-title":"Opt. Lett."},{"key":"ref_56","first-page":"894208","article-title":"Improvement of tissue analysis and classification using optical coherence tomography combined with Raman spectroscopy","volume":"8942","author":"Liu","year":"2014","journal-title":"Proc. SPIE Dyn. Fluct. Biomed. Photonics XI"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"5200108","DOI":"10.1109\/JSTQE.2015.2496265","article-title":"Simultaneous holographic microscopy and Raman spectroscopy monitoring of human spermatozoa photodegradation","volume":"22","author":"Ferrara","year":"2016","journal-title":"IEEE J. Sel. Top. Quant."},{"key":"ref_58","first-page":"9876063","article-title":"Combined Raman spectroscopy and digital holographic microscopy for sperm cell quality analysis","volume":"2017","author":"Ferrara","year":"2017","journal-title":"J. Spectrosc."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"43631","DOI":"10.1038\/srep43631","article-title":"Multimodal discrimination of immune cells using a combination of Raman spectroscopy and digital holographic microscopy","volume":"7","author":"McReynolds","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"4823","DOI":"10.1038\/s41598-019-41400-0","article-title":"Combined Raman and polarization sensitive holographic imaging for a multimodal label-free assessment of human sperm function","volume":"9","author":"Ferrara","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_61","unstructured":"Coppola, G., Zito, G., De Luca, A.C., and Ferrara, M.A. Polarized digital holography as valuable analytical toll in biological and medical research. Digit. Hologr. Three Dimens. Imaging."},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"2710","DOI":"10.1021\/acs.analchem.8b04393","article-title":"Single-cell screening of tamoxifen abundance and effect using mass spectrometry and Raman spectroscopy","volume":"91","author":"Ali","year":"2019","journal-title":"Anal. Chem."},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"929","DOI":"10.1080\/14737159.2019.1659727","article-title":"Advancing cancer diagnostics with artificial intelligence and spectroscopy: Identifying chemical changes associated with breast cancer","volume":"19","author":"Talari","year":"2019","journal-title":"Expert Rev. Mol. Diagn."}],"container-title":["Sensors"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1424-8220\/20\/19\/5525\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T10:14:05Z","timestamp":1760177645000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1424-8220\/20\/19\/5525"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2020,9,27]]},"references-count":63,"journal-issue":{"issue":"19","published-online":{"date-parts":[[2020,10]]}},"alternative-id":["s20195525"],"URL":"https:\/\/doi.org\/10.3390\/s20195525","relation":{},"ISSN":["1424-8220"],"issn-type":[{"value":"1424-8220","type":"electronic"}],"subject":[],"published":{"date-parts":[[2020,9,27]]}}}