Abstract

The investigation of living cells at physiological conditions requires very sensitive, sophisticated, non invasive methods. In this study, Raman spectral imaging is used to identity different biomolecules inside of cells. Raman spectroscopy, a chemically and structurally sensitive measuring technique, is combined with high resolution confocal microscopy. In Raman spectral imaging mode, a complete Raman spectrum is recorded at every confocal image point, giving insight into the chemical composition of each sample compartment. Neuroblastoma is the most common solid extra-cranial tumor in children. One of the unique features of neuroblastoma cells is their ability to differentiate spontaneously, eventually leading to complete remission. Since differentiation agents are currently used in the clinic for neuroblastoma therapy, there is a special need to develop non-invasive and sensitive new methods to monitor neuroblastoma cell differentiation. Neuroblastoma cells at different degrees of differentiation were analysed with the confocal Raman microscope alpha300 R (WITec GmbH, Germany), using a frequency doubled Nd:YAG laser at 532 nm and 10 mW for excitation. Integration time per spectrum was 80–100 ms. A lateral resolution in submicrometer range was achieved by using a 60x water immersion lens with a numerical aperture of 1,0. Raman images of cells were generated from these sets of data by either integrating over specific Raman bands, by basis analysis using reference spectra or by cluster analysis. The automated evaluation of all spectra results in spectral unmixed images providing insight into the chemical composition of the sample. With these procedures, different cell organelles, cytosol, membranes could be distinguished. Since neuroblastoma cells at high degree of differentiation overproduce noradrenaline, an attempt was made to trace the presence of this neurotransmitter as a marker for differentiation. The results of this work may have applications in the monitoring of molecular changes and distribution of biomolecules and in particular of low molecular weight markers as they occur during the differentiation of neuroblastoma cells.

© 2007 SPIE

PDF Article
More Like This
Raman micro-spectroscopy investigation on the effects of X-rays and polyphenols in human neuroblastoma cells

Valerio Ricciardi, Giuseppe Perna, Maria Lasalvia, Ines Delfino, Lorenzo Manti, Simona Piccolella, Severina Pacifico, Vito Capozzi, and Maria Lepore
11073_35 European Conference on Biomedical Optics (ECBO) 2019

Label-Free Detection of Tumor Markers in a Colon Carcinoma Tumor Progression Model by Confocal Raman Microspectroscopy

Claudia Scalfi-Happ, Angelika Rück, Martin Udart, Carmen Hauser, Christine Dürr, and Martin Kriebel
87980B European Conference on Biomedical Optics (ECBO) 2013

Slit-scanning Confocal Raman Microscopy: Practical Applications in Live Cell Imaging

Almar Palonpon, Masaya Okada, Jun Ando, Hiroyuki Yamakoshi, Kosuke Dodo, Mikiko Sodeoka, Satoshi Kawata, and Katsumasa Fujita
C629 Conference on Lasers and Electro-Optics/Pacific Rim (CLEOPR) 2011

References

You do not have subscription access to this journal. Citation lists with outbound citation links are available to subscribers only. You may subscribe either as an Optica member, or as an authorized user of your institution.

Contact your librarian or system administrator
or
Login to access Optica Member Subscription