Abstract
Malignancies are still responsible for a large share of lethalities. Macroscopical evaluation of the surgical resection margins is uncertain. Big data based imaging approaches have emerged in the recent decade (mass spectrometry, two-photon microscopy, infrared and Raman spectroscopy). Indocianine green labelled MS is the most common approach, however, label free mid-infrared imaging is more promising for future practical application. We aimed to identify and separate different transformed (A-375, HT-29) and non-transformed (CCD986SK) cell lines by a label-free infrared spectroscopy method. Our approach applied a novel set-up for label-free mid-infrared range classification method. Transflection spectroscopy was used on aluminium coated glass slides. Both whole range spectra (4000–648 cm−1) and hypersensitive fingerprint regions (1800–648 cm−1) were tested on the imaged areas of cell lines fixed in ethanol. Non-cell spectra were possible to be excluded based on mean transmission values being above 90%. Feasibility of a mean transmission based spectra filtering method with principal component analysis and linear discriminant analysis was shown to separate cell lines representing different tissue types. Fingerprint region resulted the best separation of cell lines spectra with accuracy of 99.84% at 70–75 mean transmittance range. Our approach in vitro was able to separate unique cell lines representing different tissues of origin. Proper data handling and spectra processing are key steps to achieve the adaptation of this dye-free technique for intraoperative surgery. Further studies are urgently needed to test this novel, marker-free approach.
Highlights
Malignancies represent a huge burden on the society and the costs of novel oncological therapies are ever increasing [1]
In the operating rooms the routine pathology background is not available on site, there is an urgent intraoperative clinical need to gain information on resected tissues, especially regarding the margins. In such cases intraoperative questions are raised to be answered by the pathologist who is situated in another location or building to where fresh tissue samples must be presented for histopathology
Our novel method used cancer cell line MIR spectra, which were pre-processed with atmospheric correction and noise reduction and were divided into 6 groups according to their mean transmittance (T%) value, calculated from the whole 4000–648 cm−1 range
Summary
Malignancies represent a huge burden on the society and the costs of novel oncological therapies are ever increasing [1]. In the operating rooms the routine pathology background is not available on site, there is an urgent intraoperative clinical need to gain information on resected tissues, especially regarding the margins In such cases intraoperative questions are raised to be answered by the pathologist who is situated in another location or building to where fresh tissue samples must be presented for histopathology. Frozen sections are cut and stained in the pathology department, which procedure takes about 15–30 min while the surgeons are waiting for answers in order to reach clinical decision This procedure lengthens the operation time and the frozen sections deliver less precise morphology due to the lack of proper dehydration, which increases the uncertainty of diagnoses
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