Implementation of NSOM to Biological Samples

Prauzner-Bechcicki, Szymon; Wiltowska-Zuber, Joanna; Budkowski, Andrzej; Lekka, Małgorzata; Rysz, Jakub

doi:10.12693/aphyspola.121.533

Cited by 4 publications

(4 citation statements)

References 16 publications

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“…Imaging of cells with subwavelength resolution in their native environment, as we performed here, is a crucial step in further studies on living cells functions and their response to various agents. However, the SNOM alignment in liquid environment is not trivial and requires some experience, because the interaction between the probe and the measured surface is difficult to control .…”

Section: Resultsmentioning

confidence: 99%

“…An alternative is the scanning near‐field optical microscopy (SNOM), which can provide extra, optical information, not available by AFM or other standard techniques . The SNOM method combines scanning approach with a classic optical microscopy, so it offers the possibility to acquire simultaneously topographical and optical images . Due to different absorbance of various substances, the SNOM offers also the chemical contrast information .…”

Section: Introductionmentioning

confidence: 99%

“…Recently, SNOM has been applied in material science and for biological samples measurement, where both the transmission and reflection SNOM configuration can be used . This method seems to be a very efficient for nanometric exploration of tissue sections, chromosomes, erythrocytes, biological membranes as well as cells .…”

Section: Introductionmentioning

confidence: 99%

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Live endothelial cells imaged by Scanning Near‐field Optical Microscopy (SNOM): capabilities and challenges

Bulat

Ryguła²,

Szafraniec

et al. 2016

Journal of Biophotonics

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The scanning near-field optical microscopy (SNOM) shows a potential to study details of biological samples, since it provides the optical images of objects with nanometric spatial resolution (50-200 nm) and the topographic information at the same time. The goal of this work is to demonstrate the capabilities of SNOM in transmission configuration to study human endothelial cells and their morphological changes, sometimes very subtle, upon inflammation. Various sample preparations were tested for SNOM measurements and promising results are collected to show: 1) the influence of α tumor necrosis factor (TNF-α) on EA.hy 926 cells (measurements of the fixed cells); 2) high resolution images of various endothelial cell lines, i.e. EA.hy 926 and HLMVEC (investigations of the fixed cells in buffer environment); 3) imaging of live endothelial cells in physiological buffers. The study demonstrate complementarity of the SNOM measurements performed in air and in liquid environments, on fixed as well as on living cells. Furthermore, it is proved that the SNOM is a very useful method for analysis of cellular morphology and topography. Changes in the cell shape and nucleus size, which are the symptoms of inflammatory reaction, were noticed in TNF-α activated EA.hy 926 cells. The cellular structures of submicron size were observed in high resolution optical images of cells from EA.hy 926 and HLMVEC lines.

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Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

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Live endothelial cells imaged by Scanning Near‐field Optical Microscopy (SNOM): capabilities and challenges

Bulat

Ryguła²,

Szafraniec

et al. 2016

Journal of Biophotonics

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“…The TOF-SIMS experiment requires a special treatment of samples, due to vacuum conditions, needed during the measurements . Cells, previously cultured on silicon surfaces, were chemically fixed using paraformaldehyde followed by drying with alcohol . Briefly, after 48 h of culture, cells attached to the silicon surface were fixed with 3.7% paraformaldehyde dissolved in phosphate buffered saline (PBS, Sigma), for 15 min in the CO 2 incubator (NuAire).…”

Section: Experimental Sectionmentioning

confidence: 99%

Differentiation between Single Bladder Cancer Cells Using Principal Component Analysis of Time-of-Flight Secondary Ion Mass Spectrometry

et al. 2015

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Time-of-flight-secondary ion mass spectrometry (TOF-SIMS) mass spectra measurements combined with an appropriate sample preparation protocol are the powerful tools to obtain unique information about the chemical composition of biological materials. In our studies, two questions were addressed, i.e., whether it is possible to develop a fixative-based sample preparation protocol and whether it allows one to distinguish between cells originating from various stages of cancer progression. Therefore, four human bladder cancer cell lines (with distinct malignancy degree) have been investigated. A chemical fixation protocol has been used for TOF-SIMS measurements, and mass spectra were obtained using a Bi3(+) primary ion beam. The principal component analysis (PCA) has been applied to analyze the whole range of mass spectra (without preselection of any particular masses) using two approaches of data preprocessing, namely, mean centering and autoscaling. The PC3 versus PC2 plot has showed significant differences between nonmalignant cancer cells and the cancerous ones for both of preprocessing approaches. The analysis of mass spectra of human bladder cells allows one to find a list of mass peaks with intensities significantly larger in cancerous bladder cells compared to nonmalignant cell cancer of the ureter (HCV29 cells). These findings show that TOF-SIMS in combination with PCA can be used to identify reference, human bladder cells from cancerous ones.

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Patterning of cancerous cells driven by a combined modification of mechanical and chemical properties of the substrate

Raczkowska

Awsiuk

Prauzner-Bechcicki

et al. 2017

European Polymer Journal

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Implementation of NSOM to Biological Samples

Cited by 4 publications

References 16 publications

Live endothelial cells imaged by Scanning Near‐field Optical Microscopy (SNOM): capabilities and challenges

Live endothelial cells imaged by Scanning Near‐field Optical Microscopy (SNOM): capabilities and challenges

Differentiation between Single Bladder Cancer Cells Using Principal Component Analysis of Time-of-Flight Secondary Ion Mass Spectrometry

Patterning of cancerous cells driven by a combined modification of mechanical and chemical properties of the substrate

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