HOT Paper!

Our article on Highlighting a need to distinguish cell cycle signatures from cellular responses to chemotherapeutics in SR-FTIR spectroscopy by Caryn, Frank, Geri and Peter, together with Mick Brown and Noel Clarke from the Paterson Institute for Cancer Research and Kevin Flower from the School of Chemistry, has been awarded Hot Paper status by Analyst.

The article is free to read until 30 November 2012.

Highlighting a need to distinguish cell cycle signatures from cellular responses to chemotherapeutics in SR-FTIR spectroscopy
C. Hughes, M. D. Brown, F. J. Ball, G. Monjardez, N. W. Clarke, K. R. Flower and P. Gardner
Analyst, 2012, 137, 5736-5742
DOI: 10.1039/C2AN35633C


Paper – Rare sub-variants of bladder cancer

FTIR microspectroscopy of selected rare diverse sub-variants of carcinoma of the urinary bladder
Caryn Hughes, Junaid Iqbal-Wahid, Michael Brown, Jonathan H. Shanks, Amanda Eustace, Helen Denley, Peter J. Hoskin, Catharine West, Noel W. Clarke, Peter Gardner
Journal of Biophotonics (2012)


Urothelial carcinomas of the bladder are a heterogeneous group of tumours, although some histological sub-variants are rare and sparsely reported in the literature. Diagnosis of sub-variants from conventional urothelial carcinoma can be challenging, as they may mimic the morphology of other malignancies or benign tumours and therefore their distinction is important. For the first time, the spectral pathology of some of these sub-variants has been documented by infrared microspectroscopy and an attempt made to profile their biochemistry. It is important not only to identify and separate the cancer-associated epithelial tissue spectra from common tissue features such as stroma or blood, but also to detect the signatures of tumour sub-variants. As shown, their spectroscopic signals can change dramatically as a consequence of differentiation. Example cases are discussed and compared with histological evaluations. (© 2012 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

Paper – Problems with transflection-mode IR

The inherent problem of transflection-mode infrared spectroscopic microscopy and the ramifications for biomedical single point and imaging applications
Paul Bassan, Joe Lee, Ashwin Sachdeva, Juliana Pissardini, Konrad M. Dorling, John S. Fletcher, Alex Henderson and Peter Gardner
, 2013, 138, 144-157


Transflection-mode FTIR spectroscopy has become a popular method of measuring spectra from biomedical and other samples due to the relative low cost of substrates compared to transmission windows, and a higher absorbance due to a double pass through the same sample approximately doubling the effective path length. In this publication we state an optical description of samples on multilayer low-e reflective substrates. Using this model we are able to explain in detail the so-called electric-field standing wave effect and rationalise the non-linear change in absorbance with sample thickness. The ramifications of this non-linear change, for imaging and classification systems, where a model is built from tissue sectioned at a particular thickness and compared with tissue of a different thickness are discussed. We show that spectra can be distorted such that classification fails leading to inaccurate tissue segmentation which may have subsequent implications for disease diagnostics applications.

Paper – Cell cycle using a synchrotron

Highlighting a need to distinguish cell cycle signatures from cellular responses to chemotherapeutics in SR-FTIR spectroscopy

C. Hughes, M. D. Brown, F. J. Ball, G. Monjardez, N. W. Clarke, K. R. Flower and P. Gardner

Analyst 137 (2012) 5736-5742


Previous research has seen difficulties in establishing clear discrimination by principal component analysis (PCA) between drug-treated cells analysed by single point SR-FTIR spectroscopy, relative to multisampling cell monolayers by conventional FTIR. It is suggested that the issue arises due to signal mixing between cellular-response signatures and cell cycle phase contributions in individual cells. Consequently, chemometric distinction of cell spectra treated with multiple drugs is difficult even with supervised methods. In an effort to separate cell cycle chemistry from cellular response chemistry in the spectra, renal carcinoma cells were stained with propidium iodide and fluorescent-activated cell sorted (FACS) after exposure to a number of chemotherapeutic compounds; 5-fluorouracil (5FU) and a set of novel gold-based experimental compounds. The cell spectra were analysed separately by PCA in G1, S or G2/M phase. The mode of action of established drug 5FU, known to disrupt S phase, was confirmed by FACS analysis. The chemical signature of 5FU-treated cells discriminated against both the control and gold-compound (KF0101)-treated cell spectra, suggesting a different mode of action due to a difference in cellular response.

Paper – Synchrotron IR for chemotherapeutics in renal cell carcinoma

Investigating cellular responses to novel chemotherapeutics in renal cell carcinoma using SR-FTIR spectroscopy
C. Hughes, M. D. Brown, N. W. Clarke, K. R. Flower and P. Gardner
Analyst 137 (2012) 4720-4726


SR-FTIR spectroscopy was evaluated as a technique to discriminate spectral signals of cellular response at the single cell level, when cancer cells are exposed to chemotherapeutics. 5-Fluorouracil, an established drug of known mode of action, was tested against a renal carcinoma cell line (Caki-2), along with two experimental analogues of gold-based compounds. The use of unsupervised principal component analysis (PCA) failed to clearly define any distinction between control and drug treated cell spectra. Supervised principal component linear discriminant analysis (PC-LDA) did have some potential to reveal signatures of cell response and repair but again failed to distinctly discriminate groups of spectra with different drug treatments. Alternatively, clear PCA discrimination was observed in spectra from average cell populations via single point benchtop spectroscopy, probing several cells simultaneously with an increased aperture. The Caki-2 cell line initially appeared to be sensitive to the novel compounds, inducing a cellular response prior to subsequential cell recovery which was assessed by both PCA and cell viability assays.


Welcome to the website of the Gardner Lab. The group is headed by Professor Peter Gardner who has 25 years of research experience in the field of vibrational spectroscopy. For the last 10 years the group has focused on the development of vibrational and related spectroscopies for biomedical applications with a particular interest in cancer diagnosis.

Here you can find out more about group members, the group’s activities, current research, and PhD studentships. There are also conference reports and news items that we will try and keep up to date.

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