Beata Brozek-Pluska

Phase transitions in oleic acid and in human breast tissue as studied by Raman spectroscopy and Raman imaging.

We present the results of differential scanning calorimetry (DSC) and Raman studies in the temperature range of 293-77 K on vibrational properties of the oleic acid and the human breast tissue as a function of temperature. We have found that vibrational properties are very sensitive indicators to specify phases and phase transitions at the molecular level. We have found that water content confined in the cancerous tissue is markedly different from that in the noncancerous tissue. The OH stretching vibrations of water are useful as potential Raman biomarkers to distinguish between the cancerous and the noncancerous human breast tissues. Our results provide experimental evidence on the role of lipid profile and cell hydration as factors of particular significance in differentiation of the noncancerous and cancerous breast tissues.

New look inside human breast ducts with Raman imaging. Raman candidates as diagnostic markers for breast cancer prognosis: Mammaglobin, palmitic acid and sphingomyelin.

Looking inside the human body fascinated mankind for thousands of years. Current diagnostic and therapy methods are often limited by inadequate sensitivity, specificity and spatial resolution. Raman imaging may bring revolution in monitoring of disease and treatment. The main advantage of Raman imaging is that it gives spatial information about various chemical constituents in defined cellular organelles in contrast to conventional methods (liquid chromatography/mass spectrometry, NMR, HPLC) that rely on bulk or fractionated analyses of extracted components. We demonstrated how Raman imaging can drive the progress on breast cancer just unimaginable a few years ago. We looked inside human breast ducts answering fundamental questions about location and distribution of various biochemical components inside the lumen, epithelial cells of the duct and the stroma around the duct during cancer development. We have identified Raman candidates as diagnostic markers for breast cancer prognosis: carotenoids, mammaglobin, palmitic acid and sphingomyelin as key molecular targets in ductal breast cancer in situ, and propose the molecular mechanisms linking oncogenes with lipid programming.

Human breast tissue cancer diagnosis by Raman spectroscopy

Differences between Raman spectra of normal, malignant and benign tissues have been recorded and analyzed as a method for the early detection of cancer. To the best of our knowledge, this is one of the most statistically reliable research (67 patients) on Raman spectroscopy-based diagnosis of breast cancers among the world women population. The paper demonstrates that Raman spectroscopy is a promising new tool for real-time diagnosis of tissue abnormalities.

Distribution of phthalocyanines and Raman reporters in human cancerous and noncancerous breast tissue as studied by Raman imaging.

There is a considerable interest in the developing new diagnostic techniques allowing noninvasive tracking of the progress of therapies used to treat a cancer. Raman imaging of distribution of phthalocyanine photosensitizers may open new possibilities of Photodynamic Therapy (PDT) to treat a wide range of neoplastic lesions with improved effectiveness of treatment through precise identification of malignant areas. We have employed Raman imaging and Raman spectroscopy to analyze human breast cancer tissue that interacts with photosensitizers used in the photodynamic therapy of cancer. PCA (Principal Component Analysis) has been employed to analyze various areas of the noncancerous and cancerous breast tissues. The results show that the emission spectra combined with the Raman images are very sensitive indicators to specify the aggregation state and the distribution of phthalocyanines in the cancerous and noncancerous breast tissues. Our results provide experimental evidence on the role of aggregation of phthalocyanines as a factor of particular significance in differentiation of the normal and tumourous (cancerous or benign pathology) breast tissues. We conclude that the Raman imaging reported here has a potential to be a novel and effective photodynamic therapeutic method with improved selectivity for the treatment of breast cancer.

Development of a new diagnostic Raman method for monitoring epigenetic modifications in the cancer cells of human breast tissue

Cancer diagnosis requires better screening of early stages of pathology and monitoring patient responses to treatment. Current technologies in the clinical sector are expensive, sophisticated and time consuming. This paper develops a novel Raman based alternative for currently existing epigenetics research approaches. The proposed Raman approach can ‘upgrade’ cancer epigenetic tests and answer many questions by monitoring the biochemistry of cancer cells. We will show that Raman spectroscopy and Raman imaging can detect the relative amounts of acetylated and methylated lysine by monitoring the vibrations of the acetyl and methyl chemical functional groups. In comparison to existing tests and assays used to monitor molecular processes of acetylation and methylation of all proteins, Raman-based methods have the potential to be a powerful alternative for conventional methods of cell biology, because they allow non-invasive detection of cellular acetylation and methylation processes that are not limited to only those events that are sensitive to a specific antibody. Vibrational Raman signatures of acetylation and methylation processes in epithelial cells of human breast tissue (ductal and lobular carcinoma) have been used to identify and discriminate structures in normal and cancerous tissues. Our results demonstrate that the stretching vibration of the acetyl group observed at around 2938–2942 cm−1 and the methyl group at 2970 cm−1 by Raman spectroscopy is useful in monitoring these epigenetic molecular processes in cancer cells. The Raman vibrational marker is markedly blue-shifted from 2905 cm−1 (non-acetylated C–H vibrations) for normal cells to 2942 cm−1 (vibrations of the acetylated functional group) in cancer cells. The sensitivity and specificity obtained directly from PLSDA and cross-validation for a chosen model gives a sensitivity and specificity of 86.1% and 91.3% for calibration and 85.3% and 91.3% for cross-validation, respectively. The results presented in the paper provide strong evidence that the global acetylation level of histone and non-histone proteins increases in human breast cancer cells.

Phthalocyanines: From Dyes to Photosensitizers in Diagnostics and Treatment of Cancer. Spectroscopy and Raman Imaging Studies of Phthalocyanines in Human Breast Tissues

1 Lodz University of Technology, Faculty of Chemistry, Institute of Applied Radiation Chemistry, Laboratory of Laser Molecular Spectroscopy, Wroblewskiego 15, 93-590 Lodz, Poland. 2 Lodz University of Technology, Faculty of Electrical, Electronic, Computer and Control Engineering, Institute of Electrical Power Engeneering, Instrument Transformers and EMC Division, Stefanowskiego 18/22, 90-924 Lodz, Poland.

Surface-Enhanced Raman Spectroscopy Analysis of Human Breast Cancer via Silver Nanoparticles: An Examination of Fabrication Methods

Breast cancer in a traditional way is diagnosed using mammography, computer tomography, ultrasounds, biopsy, and finally, histopathological analysis. Histopathological analysis is a gold standard in breast cancer diagnostics; however, it is time consuming and prone to the human interpretations. That is why new methods based on optical properties of analyzed human tissue samples are needed to be introduced to the clinical practice objective, costless and fast diagnostic protocols. Nowadays, Raman spectroscopy-based methods are gaining more and more importance. Raman spectroscopy and imaging allow to characterize human tissue samples using an electromagnetic radiation from a safe range, and simultaneously, a minimal sample preparation is required. During measurements, a natural differentiation in tissues components’ scattering cross sections is used to build 2D and 3D maps of the chemical component distribution. The paper presents the application of SERS (surface-enhanced Raman spectroscopy) measurements for analysis of human breast cancer (adenocarcinoma). The advantages of SERS application in cancer diagnostics are also discussed. Moreover, the detailed chemical composition of human breast cancer tissue based on Raman bands of DNA/RNA, amino acids, lipids, and proteins which are significantly enhanced is presented. Three different methods of NP preparation are presented, and the effectiveness of Raman signal enhancement of Ag nanoparticles synthetized by these methods is compared. The enhancement effect of NPs synthetized by reduction of silver nitrate with sodium borohydride (method no. 1) and silver nitrate-hydroxylamine hydrochloride reduction (method no. 2) was stronger when compared with the polyol method (method no. 3). Presented SERS results confirmed that the clearly resolved and high-intensity Raman spectra of cancer human breast tissue can be recorded using integration times of the order of fractional seconds and one milliwatt of the excitation laser power.

From breast tissue diagnosis by Raman spectroscopy to femtosecond dynamics at the phospholipid membrane-water interface

In this paper we present our recent results on normal, malignant, and benign breast tissue by Raman spectroscopy for 100 patients and compare them with the results of histopathological analysis to estimate the sensitivity and specificity of the method. The typical Raman spectra and microscope picture of the sample prepared for histopatological analysis of the normal and pathological tissues are presented in Fig. 1.