Colocalization of cellular nanostructure using confocal fluorescence and partial wave spectroscopy

A new multimodal confocal microscope has been developed, which includes a parallel Partial Wave Spectroscopic (PWS) microscopy path. This combination of modalities allows molecular‐specific sensing of nanoscale intracellular structure using fluorescent labels. Combining molecular specificity and sen...

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Bibliographic Details
Published in:Journal of biophotonics Vol. 10; no. 3; pp. 377 - 384
Main Authors: Chandler, John E., Stypula‐Cyrus, Yolanda, Almassalha, Luay, Bauer, Greta, Bowen, Leah, Subramanian, Hariharan, Szleifer, Igal, Backman, Vadim
Format: Journal Article
Language:English
Published: Weinheim WILEY‐VCH Verlag 01.03.2017
Wiley Subscription Services, Inc
Subjects:
Cancer
Cancer screening
Cell Nucleus - drug effects
Cell Nucleus - ultrastructure
Cheek
chromatin
Confocal
confocal microscopy
Fluorescence
HeLa Cells
Heterogeneity
Humans
hyperspectral microscopy
Image Processing, Computer-Assisted
Intracellular
Ionophores - pharmacology
Localization
Medical imaging
Medical screening
Microscopy, Confocal - instrumentation
Microscopy, Confocal - methods
Microscopy, Fluorescence - instrumentation
Microscopy, Fluorescence - methods
Mitochondria
Mitochondria - drug effects
Mitochondria - ultrastructure
Molecular structure
Multimodal Imaging
nanocytology
Nanostructure
Oxidative phosphorylation
Phosphorylation
Spectroscopy
Spectrum Analysis - instrumentation
Spectrum Analysis - methods
Valinomycin
Valinomycin - pharmacology
hyperspectral microscopy
chromatin
confocal microscopy
nanocytology
mitochondria
ISSN:1864-063X, 1864-0648, 1864-0648
Online Access:Get full text
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Summary:A new multimodal confocal microscope has been developed, which includes a parallel Partial Wave Spectroscopic (PWS) microscopy path. This combination of modalities allows molecular‐specific sensing of nanoscale intracellular structure using fluorescent labels. Combining molecular specificity and sensitivity to nanoscale structure allows localization of nanostructural intracellular changes, which is critical for understanding the mechanisms of diseases such as cancer. To demonstrate the capabilities of this multimodal instrument, we imaged HeLa cells treated with valinomycin, a potassium ionophore that uncouples oxidative phosphorylation. Colocalization of fluorescence images of the nuclei (Hoechst 33342) and mitochondria (anti‐mitochondria conjugated to Alexa Fluor 488) with PWS measurements allowed us to detect a significant decrease in nuclear nanoscale heterogeneity (Σ), while no significant change in Σ was observed at mitochondrial sites. In addition, application of the new multimodal imaging approach was demonstrated on human buccal samples prepared using a cancer screening protocol. These images demonstrate that nanoscale intracellular structure can be studied in healthy and diseased cells at molecular‐specific sites. Instrumentation has been developed enabling colocalization of confocal fluorescence and nanoscale‐sensitive spectroscopic microscopy modalities. This combination allows sensing of the heterogeneity of nanoscale intracellular structure within specific fluorescently labeled organelles or molecular structures. To demonstrate the capabilities of this multimodal instrument, the effects on the heterogeneity of nanoscale structure were examined in the nuclei and mitochondria of HeLa cells treated with valinomycin, a potassium ionophore that uncouples oxidative phosphorylation.
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ISSN:1864-063X
1864-0648
1864-0648
DOI:10.1002/jbio.201500298