Skip to main page content
Access keys NCBI Homepage MyNCBI Homepage Main Content Main Navigation
. 2013 Nov;18(11):117008.
doi: 10.1117/1.JBO.18.11.117008.

Differentiation of Cancer Cells in Two-Dimensional and Three-Dimensional Breast Cancer Models by Raman Spectroscopy

Affiliations
Free PMC article

Differentiation of Cancer Cells in Two-Dimensional and Three-Dimensional Breast Cancer Models by Raman Spectroscopy

Nur P Damayanti et al. J Biomed Opt. .
Free PMC article

Abstract

We demonstrate the first application of Raman spectroscopy in diagnosing nonmalignant, premalignant, malignant, and metastatic stages of breast cancer in a three-dimensional (3-D) cell culture model that closely mimics an in vivo environment. Comprehensive study comparing classification in two-dimensional (2-D) and 3-D cell models was performed using statistical methods composed of principal component analysis for exploratory analysis and outlier removal, partial least squares discriminant analysis, and elastic net regularized regression for classification. Our results show that Raman spectroscopy with an appropriate classification tool has excellent resolution to discriminate the four stages of breast cancer progression, with a near 100% accuracy for both 2-D and 3-D cell models. The diversity in chemical groups related to nucleic acids, proteins, and lipids, among other chemicals, were identified by appropriate peaks in the Raman spectra that correspond to the correct classification of the different stages of tumorigenesis model comprising of MCF10A, MCF10AneoT, MCF10CA1h, and MCF10CA1a cell lines. An explicit relationship between wavenumber and the stages of cancer progression was identified by the elastic net variable selection.

Figures

Fig. 1
Fig. 1
Schematic of a two-dimensional (2-D) and three-dimensional (3-D) cell culture systems: (a) 2-D; (b and c) 3-D culture models (top) and 3-D cell sample preparation (bottom). 3-D cell lines were first cultured in a 24-well plate in an embedded model, washed, and a piece of gel was then taken, placed onto the gold slide for Raman measurement that followed.
Fig. 2
Fig. 2
Schematic of sampling for 2-D and 3-D culture systems: (a) five spots were chosen for data collection in each 2-D cell, (b) 15 spots distributed throughout the spheroid were chosen in the 3-D spheroid, and (c) schematic of the optical layout of the Senterra Confocal Raman spectrometer: the laser is directed through a neutral density filter to excite the sample through the objective and the scattered photons are collected by the same objective. A pinhole is used to detect photons from the plane of focus.
Fig. 3
Fig. 3
Mean Raman spectra of (a) 2-D and (b) 3-D cell cultures of benign, MCF10A (black), premalignant, MCF10AneoT (dark gray), malignant, MCF10CA1h (medium gray), and metastatic MCF10CA1h (light gray). Standard deviation is provided.
Fig. 4
Fig. 4
Three-dimensional cell culture model better discriminates the four stages of breast cancer; PC scores scatter plots: (a), (c), PC1 versus PC2 of 2-D (a) and 3-D (c) cell models, (b), (d), PC1versus PC2 versus PC3 of (b) 2-D, and (d) 3-D models.
Fig. 5
Fig. 5
Elastic net coefficient estimates for α=0.5 for (a) 2-D and (b) 3-D cell culture models. A peak indicates that the correct classification of spectra is associated with the corresponding spectral region. A positive peak indicates higher intensity than other cell lines; a negative peak indicates lower intensity.

Similar articles

See all similar articles

Cited by 4 articles

Publication types

Feedback