Document Type



1.6 BIOLOGICAL SCIENCES, 3. MEDICAL AND HEALTH SCIENCES, Radiology, nuclear medicine and medical imaging

Publication Details

Monaghan JF, Cullen D, Wynne C, Lyng FM, Meade AD. Effect of pre-analytical variables on Raman and FTIR spectral content of lymphocytes. Analyst. 2023 Oct 23;148(21):5422-5434.

doi: 10.1039/d3an00686g.


The use of Fourier transform infrared (FTIR) and Raman spectroscopy (RS) for the analysis of lymphocytes in clinical applications is increasing in the field of biomedicine. The pre-analytical phase, which is the most vulnerable stage of the testing process, is where most errors and sample variance occur; however, it is unclear how pre-analytical variables affect the FTIR and Raman spectra of lymphocytes. In this study, we evaluated how pre-analytical procedures undertaken before spectroscopic analysis influence the spectral integrity of lymphocytes purified from the peripheral blood of male volunteers (n = 3). Pre-analytical variables investigated were associated with (i) sample preparation, (blood collection systems, anticoagulant, needle gauges), (ii) sample storage (fresh or frozen), and (iii) sample processing (inter-operator variability, time to lymphocyte isolation). Although many of these procedural pre-analytical variables did not alter the spectral signature of the lymphocytes, evidence of spectral effects due to the freeze–thaw cycle, in vitro culture inter-operator variability and the time to lymphocyte isolation was observed. Although FTIR and RS possess clinical potential, their translation into a clinical environment is impeded by a lack of standardisation and harmonisation of protocols related to the preparation, storage, and processing of samples, which hinders uniform, accurate, and reproducible analysis. Therefore, further development of protocols is required to successfully integrate these techniques into current clinical workflows.



J. F. M. is funded by Fiosraigh Enterprise Award co-funded by TU Dublin and St Luke’s Institute for Cancer Research. D. C. was funded by a TU Dublin Fiosraigh scholarship.

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Creative Commons Attribution-Share Alike 4.0 International License
This work is licensed under a Creative Commons Attribution-Share Alike 4.0 International License.