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Fluorescent Probes for Intracellular Imaging and Super Resolution Microscopy

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A special issue of Sensors (ISSN 1424-8220). This special issue belongs to the section "Chemical Sensors".

Deadline for manuscript submissions: closed (31 August 2015) | Viewed by 32110

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Special Issue Editors

Prof. Dr. Frances S. Ligler

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Guest Editor

Lampe Distinguished Professor, Joint Department of Biomedical Engineering, University of North Carolina-Chapel Hill and North Carolina State University, NCSU Engineering Building 3, Room 4307 Mail Stop 7115, Raleigh, NC 27695-7115, USA
Interests: biosensors; immunosensors; fluorescence; multi-analyte sensing; microfluidics; flow cytometry; microarrays
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Dr. Brandy J. Johnson

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Center for Biomolecular Science and Engineering, Code 6900, US Naval Research Laboratory, Washington, DC 20375, USA
Interests: protective coatings; distributed sensing; environmental monitoring; self-decontaminating materials, and natural products
Special Issues, Collections and Topics in MDPI journals

Special Issue Information

Dear Colleagues,

A number of microscopy approaches have been developed that provide the potential for investigating processes and nanostructures within cells and tissues. Innovative methods that provide information on real-time changes in concentrations of ions inside cells (e.g. NO, Mg, Ca), enzyme activity, or structural elements are being developed. Of particular interest are measurements at resolutions below the light diffraction limits of traditional fluorescence microscopy. These methods include near-field scanning optical microscopy (NSOM), stimulated emission depletion (STED) microscopy, stochastic optical reconstruction microscopy (STORM), and structured illumination microscopy (SIM). This Special Issue highlights the identification and development of fluorescent probes that provide new capabilities for intracellular measurements that have not been possible in the past. The unique and specific features highlighted include novel specificity and biocompatibility, new advantages in terms of wavelength, quantum yield, and range, and unique properties that facilitate the use of super resolution microscopy, so as to provide information at resolutions as low as tens of nanometers.

Example of the previous edition:

https://www.mdpi.com/journal/sensors/special_issues/fluorescent-chemosensors

Prof. Frances S. Ligler
Dr. Brandy J. Johnson
Guest Editors

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Keywords

superresolution microscopy
sub-diffraction microscopy
ultrastructure
molecular reporters
cell imaging
tissue imaging
fluorphore
molecular probe
STED
STORM

Published Papers (4 papers)

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Research

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635 KiB

Open AccessArticle

Continuous-Wave Stimulated Emission Depletion Microscope for Imaging Actin Cytoskeleton in Fixed and Live Cells

by Bhanu Neupane, Tao Jin, Liliana F. Mellor, Elizabeth G. Loboa, Frances S. Ligler and Gufeng Wang

Sensors 2015, 15(9), 24178-24190; https://doi.org/10.3390/s150924178 - 18 Sep 2015

Cited by 10 | Viewed by 6409

Abstract

Stimulated emission depletion (STED) microscopy provides a new opportunity to study fine sub-cellular structures and highly dynamic cellular processes, which are challenging to observe using conventional optical microscopy. Using actin as an example, we explored the feasibility of using a continuous wave (CW)-STED microscope to study the fine structure and dynamics in fixed and live cells. Actin plays an important role in cellular processes, whose functioning involves dynamic formation and reorganization of fine structures of actin filaments. Frequently used confocal fluorescence and STED microscopy dyes were employed to image fixed PC-12 cells (dyed with phalloidin- fluorescein isothiocyante) and live rat chondrosarcoma cells (RCS) transfected with actin-green fluorescent protein (GFP). Compared to conventional confocal fluorescence microscopy, CW-STED microscopy shows improved spatial resolution in both fixed and live cells. We were able to monitor cell morphology changes continuously; however, the number of repetitive analyses were limited primarily by the dyes used in these experiments and could be improved with the use of dyes less susceptible to photobleaching. In conclusion, CW-STED may disclose new information for biological systems with a proper characteristic length scale. The challenges of using CW-STED microscopy to study cell structures are discussed. Full article

(This article belongs to the Special Issue Fluorescent Probes for Intracellular Imaging and Super Resolution Microscopy)

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1614 KiB

Open AccessArticle

Quadruplex Integrated DNA (QuID) Nanosensors for Monitoring Dopamine

by Jennifer M. Morales, Christopher G. Skipwith and Heather A. Clark

Sensors 2015, 15(8), 19912-19924; https://doi.org/10.3390/s150819912 - 13 Aug 2015

Cited by 8 | Viewed by 6064

Abstract

Dopamine is widely innervated throughout the brain and critical for many cognitive and motor functions. Imbalances or loss in dopamine transmission underlie various psychiatric disorders and degenerative diseases. Research involving cellular studies and disease states would benefit from a tool for measuring dopamine transmission. Here we show a Quadruplex Integrated DNA (QuID) nanosensor platform for selective and dynamic detection of dopamine. This nanosensor exploits DNA technology and enzyme recognition systems to optically image dopamine levels. The DNA quadruplex architecture is designed to be compatible in physically constrained environments (110 nm) with high flexibility, homogeneity, and a lower detection limit of 110 µM. Full article

(This article belongs to the Special Issue Fluorescent Probes for Intracellular Imaging and Super Resolution Microscopy)

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Review

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4271 KiB

Open AccessReview

Membrane Potential and Calcium Dynamics in Beta Cells from Mouse Pancreas Tissue Slices: Theory, Experimentation, and Analysis

by Jurij Dolenšek, Denis Špelič, Maša Skelin Klemen, Borut Žalik, Marko Gosak, Marjan Slak Rupnik and Andraž Stožer

Sensors 2015, 15(11), 27393-27419; https://doi.org/10.3390/s151127393 - 28 Oct 2015

Cited by 22 | Viewed by 7206

Abstract

Beta cells in the pancreatic islets of Langerhans are precise biological sensors for glucose and play a central role in balancing the organism between catabolic and anabolic needs. A hallmark of the beta cell response to glucose are oscillatory changes of membrane potential that are tightly coupled with oscillatory changes in intracellular calcium concentration which, in turn, elicit oscillations of insulin secretion. Both membrane potential and calcium changes spread from one beta cell to the other in a wave-like manner. In order to assess the properties of the abovementioned responses to physiological and pathological stimuli, the main challenge remains how to effectively measure membrane potential and calcium changes at the same time with high spatial and temporal resolution, and also in as many cells as possible. To date, the most wide-spread approach has employed the electrophysiological patch-clamp method to monitor membrane potential changes. Inherently, this technique has many advantages, such as a direct contact with the cell and a high temporal resolution. However, it allows one to assess information from a single cell only. In some instances, this technique has been used in conjunction with CCD camera-based imaging, offering the opportunity to simultaneously monitor membrane potential and calcium changes, but not in the same cells and not with a reliable cellular or subcellular spatial resolution. Recently, a novel family of highly-sensitive membrane potential reporter dyes in combination with high temporal and spatial confocal calcium imaging allows for simultaneously detecting membrane potential and calcium changes in many cells at a time. Since the signals yielded from both types of reporter dyes are inherently noisy, we have developed complex methods of data denoising that permit for visualization and pixel-wise analysis of signals. Combining the experimental approach of high-resolution imaging with the advanced analysis of noisy data enables novel physiological insights and reassessment of current concepts in unprecedented detail. Full article

(This article belongs to the Special Issue Fluorescent Probes for Intracellular Imaging and Super Resolution Microscopy)

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1476 KiB

Open AccessReview

Recent Progress in Fluorescent Imaging Probes

by Yen Leng Pak, K. M. K. Swamy and Juyoung Yoon

Sensors 2015, 15(9), 24374-24396; https://doi.org/10.3390/s150924374 - 22 Sep 2015

Cited by 100 | Viewed by 11691

Abstract

Due to the simplicity and low detection limit, especially the bioimaging ability for cells, fluorescence probes serve as unique detection methods. With the aid of molecular recognition and specific organic reactions, research on fluorescent imaging probes has blossomed during the last decade. Especially, reaction based fluorescent probes have been proven to be highly selective for specific analytes. This review highlights our recent progress on fluorescent imaging probes for biologically important species, such as biothiols, reactive oxygen species, reactive nitrogen species, metal ions including Zn²⁺, Hg²⁺, Cu²⁺ and Au³⁺, and anions including cyanide and adenosine triphosphate (ATP). Full article

(This article belongs to the Special Issue Fluorescent Probes for Intracellular Imaging and Super Resolution Microscopy)

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