Label-free non-contact imaging of cell nuclei using ultraviolet photoacoustic remote sensing microscopy (Conference Presentation)

2019 ◽  
Author(s):  
Nathaniel J. M. Haven ◽  
Kevan L. Bell ◽  
Pradyumna Kedarisetti ◽  
Parsin Haji Reza ◽  
John D. Lewis ◽  
...  
Keyword(s):  
Remote Sensing ◽  
Label Free ◽  
Cell Nuclei ◽  
Contact Imaging

F-mode ultraviolet photoacoustic remote sensing for label-free virtual H&E histopathology using a single excitation wavelength

2021 ◽  
Author(s):  
Pradyumna Kedarisetti ◽  
Brendon Restall ◽  
Nathaniel Haven ◽  
Matthew Martell ◽  
Brendyn Cikaluk ◽  
...  
Keyword(s):  
Remote Sensing ◽  
Excitation Wavelength ◽  
Label Free

Label-free virtual H&E histopathology using f-mode ultraviolet photoacoustic remote sensing

2021 ◽  
Author(s):  
Pradyumna Kedarisetti ◽  
Nathaniel J. M. Haven ◽  
Brendon S. Restall ◽  
Matthew T. Martell ◽  
Roger J. Zemp
Keyword(s):  
Remote Sensing ◽  
Label Free

scholarly journals Systematic Enzyme Mapping of Cellular Metabolism by Phasor-Analyzed Label-Free NAD(P)H Fluorescence Lifetime Imaging

2019 ◽  
Vol 20 (22) ◽  
pp. 5565 ◽  
Author(s):  
Leben ◽  
Köhler ◽  
Radbruch ◽  
Hauser ◽  
Niesner
Keyword(s):  
Fluorescence Lifetime ◽  
Metabolic Activity ◽  
Cellular Metabolism ◽  
Fluorescence Lifetime Imaging ◽  
Label Free ◽  
Analysis Framework ◽  
Cell Nuclei ◽  
Lifetime Imaging ◽  
Subcellular Resolution ◽  
In Cells

In the past years, cellular metabolism of the immune system experienced a revival, as it has become clear that it is not merely responsible for the cellular energy supply, but also impacts on many signaling pathways and, thus, on diverse cellular functions. Label-free fluorescence lifetime imaging of the ubiquitous coenzymes NADH and NADPH (NAD(P)H-FLIM) makes it possible to monitor cellular metabolism in living cells and tissues and has already been applied to study metabolic changes both under physiologic and pathologic conditions. However, due to the complex distribution of NAD(P)H-dependent enzymes in cells, whose distribution continuously changes over time, a thorough interpretation of NAD(P)H-FLIM results, in particular, resolving the contribution of various enzymes to the overall metabolic activity, remains challenging. We developed a systematic framework based on angle similarities of the phase vectors and their length to analyze NAD(P)H-FLIM data of cells and tissues based on a generally valid reference system of highly abundant NAD(P)H-dependent enzymes in cells. By using our analysis framework, we retrieve information not only about the overall metabolic activity, i.e., the fraction of free to enzyme-bound NAD(P)H, but also identified the enzymes predominantly active within the sample at a certain time point with subcellular resolution. We verified the performance of the approach by applying NAD(P)H-FLIM on a stromal-like cell line and identified a different group of enzymes that were active in the cell nuclei as compared to the cytoplasm. As the systematic phasor-based analysis framework of label-free NAD(P)H-FLIM can be applied both in vitro and in vivo, it retains the unique power to enable dynamic enzyme-based metabolic investigations, at subcellular resolution, in genuine environments.

scholarly journals Label-Free, High-Throughput Measurements of Dynamic Changes in Cell Nuclei Using Angle-Resolved Low Coherence Interferometry

2008 ◽  
Vol 94 (12) ◽  
pp. 4948-4956 ◽  
Author(s):  
Kevin J. Chalut ◽  
Sulin Chen ◽  
John D. Finan ◽  
Michael G. Giacomelli ◽  
Farshid Guilak ◽  
...  
Keyword(s):  
High Throughput ◽  
Label Free ◽  
Cell Nuclei ◽  
Dynamic Changes ◽  
Low Coherence ◽  
Low Coherence Interferometry

scholarly journals Single cell label-free probing of chromatin dynamics during B lymphocyte maturation

2021 ◽  
Author(s):  
Rikke Morrish ◽  
Kevin Ho Wai Yim ◽  
Stefano Pagliara ◽  
Francesca Palombo ◽  
Richard Chahwan ◽  
...  
Keyword(s):  
Single Cell ◽  
Cell Fate ◽  
B Lymphocytes ◽  
Large Scale ◽  
B Lymphocyte ◽  
Biological Effects ◽  
Raman Microscopy ◽  
Label Free ◽  
Cell Nuclei ◽  
Phenotypic Characterisation

ABSTRACTLarge-scale intracellular signalling during developmental growth or in response to environmental alterations are largely orchestrated by chromatin within the cell nuclei. Chemical and conformational modifications of the chromatin architecture are critical steps in the regulation of differential gene expression and ultimately cell fate determination. Therefore, establishing chemical properties of the nucleus could provide key markers for phenotypic characterisation of cellular processes on a scale of individual cells.Raman microscopy is a sensitive technique that is capable of probing single cell chemical composition - and sub-cellular regions - in a label-free optical manner. As such, it has great potential in both clinical and basic research. However, perceived limitations of Raman spectroscopy such as low signal intensity and the difficulty in linking alterations in vibrational signals directly with ensuing biological effects have hampered advances in the field. Here we use immune B lymphocyte development as a model to assess chromatin and transcriptional changes using confocal Raman microscopy in combination with microfluidic devices and correlative transcriptomics, thereby linking changes in chemical and structural properties to biological outcomes. Live B lymphocytes were assessed before and after maturation. Multivariate analysis was applied to distinguish cellular components within each cell. The spectral differences between non-activated and activated B lymphocytes were then identified, and their correlation with known intracellular biological changes were assessed in comparison to conventional RNA-seq analysis. Our data shows that spectral analysis provides a powerful tool to study gene activation that can complement conventional molecular biology techniques and opens the way for mapping the dynamics in the biochemical makeup of individual cells.

scholarly journals Label-free, non-contact, in vivo ophthalmic imaging using photoacoustic remote sensing microscopy

2020 ◽  
Vol 45 (22) ◽  
pp. 6254
Author(s):  
Zohreh Hosseinaee ◽  
Layla Khalili ◽  
James A. Tummon Simmons ◽  
Kevan Bell ◽  
Parsin Haji Reza
Keyword(s):  
Remote Sensing ◽  
Label Free ◽  
Ophthalmic Imaging

scholarly journals Single acquisition label-free histology-like imaging with dual-contrast photoacoustic remote sensing microscopy

2021 ◽  
Vol 26 (05) ◽  
Author(s):  
Benjamin Ecclestone ◽  
Deepak Dinakaran ◽  
Parsin Haji Reza
Keyword(s):  
Remote Sensing ◽  
Label Free
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