Visualizing chemical interactions in life sciences with wide-field fluorescence microscopy towards the single-molecule level

2007 ◽  
Vol 26 (10) ◽  
pp. 980-992 ◽  
Author(s):  
Hongwei Gai ◽  
Isaac Stayton ◽  
Xin Liu ◽  
Bingcheng Lin ◽  
Yinfa Ma
Keyword(s):  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Life Sciences ◽  
Wide Field ◽  
Chemical Interactions ◽  
Single Molecule Level

Direct observation of structural properties and fluorescent trapping sites in macrocyclic porphyrin arrays at the single-molecule level

2016 ◽  
Vol 18 (5) ◽  
pp. 3871-3877 ◽  
Author(s):  
Sujin Ham ◽  
Ji-Eun Lee ◽  
Suhwan Song ◽  
Xiaobin Peng ◽  
Takaaki Hori ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Structural Properties ◽  
Direct Observation ◽  
Single Molecule ◽  
Site Selection ◽  
Wide Field ◽  
Single Molecule Level ◽  
Selection For ◽  
Trapping Sites

By utilizing single-molecule defocused wide-field fluorescence microscopy, we have investigated the molecular structural properties and ascertained site selection for fluorescent trapping sites in multichromophoric systems.

scholarly journals Low-Light Photodetectors for Fluorescence Microscopy

2021 ◽  
Vol 11 (6) ◽  
pp. 2773
Author(s):  
Hiroaki Yokota ◽  
Atsuhito Fukasawa ◽  
Minako Hirano ◽  
Toru Ide
Keyword(s):  
Fluorescence Microscopy ◽  
Fluorescence Imaging ◽  
Single Molecule ◽  
Science Studies ◽  
Single Photon ◽  
Laser Scanning Microscopy ◽  
Oxide Semiconductor ◽  
Wide Field ◽  
Single Molecule Fluorescence ◽  
Low Light

Over the years, fluorescence microscopy has evolved and has become a necessary element of life science studies. Microscopy has elucidated biological processes in live cells and organisms, and also enabled tracking of biomolecules in real time. Development of highly sensitive photodetectors and light sources, in addition to the evolution of various illumination methods and fluorophores, has helped microscopy acquire single-molecule fluorescence sensitivity, enabling single-molecule fluorescence imaging and detection. Low-light photodetectors used in microscopy are classified into two categories: point photodetectors and wide-field photodetectors. Although point photodetectors, notably photomultiplier tubes (PMTs), have been commonly used in laser scanning microscopy (LSM) with a confocal illumination setup, wide-field photodetectors, such as electron-multiplying charge-coupled devices (EMCCDs) and scientific complementary metal-oxide-semiconductor (sCMOS) cameras have been used in fluorescence imaging. This review focuses on the former low-light point photodetectors and presents their fluorescence microscopy applications and recent progress. These photodetectors include conventional PMTs, single photon avalanche diodes (SPADs), hybrid photodetectors (HPDs), in addition to newly emerging photodetectors, such as silicon photomultipliers (SiPMs) (also known as multi-pixel photon counters (MPPCs)) and superconducting nanowire single photon detectors (SSPDs). In particular, this review shows distinctive features of HPD and application of HPD to wide-field single-molecule fluorescence detection.

A Single-Molecule Homogeneous Immunoassay by Counting Spatially “Overlapping” Two-Color Quantum Dots with Wide-Field Fluorescence Microscopy

ACS Sensors ◽  
2018 ◽  
Vol 3 (12) ◽  
pp. 2644-2650 ◽  
Author(s):  
Xiaojun Liu ◽  
Conghui Huang ◽  
Chenghua Zong ◽  
Aiye Liang ◽  
Zhangjian Wu ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Wide Field ◽  
Homogeneous Immunoassay

scholarly journals Development of new photon-counting detectors for single-molecule fluorescence microscopy

2013 ◽  
Vol 368 (1611) ◽  
pp. 20120035 ◽  
Author(s):  
X. Michalet ◽  
R. A. Colyer ◽  
G. Scalia ◽  
A. Ingargiola ◽  
R. Lin ◽  
...  
Keyword(s):  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Single Photon ◽  
Photon Counting ◽  
Wide Field ◽  
Fluorescence Lifetime Imaging ◽  
Large Area ◽  
Single Molecule Fluorescence ◽  
Single Photon Counting ◽  
Single Molecule Fluorescence Microscopy

Two optical configurations are commonly used in single-molecule fluorescence microscopy: point-like excitation and detection to study freely diffusing molecules, and wide field illumination and detection to study surface immobilized or slowly diffusing molecules. Both approaches have common features, but also differ in significant aspects. In particular, they use different detectors, which share some requirements but also have major technical differences. Currently, two types of detectors best fulfil the needs of each approach: single-photon-counting avalanche diodes (SPADs) for point-like detection, and electron-multiplying charge-coupled devices (EMCCDs) for wide field detection. However, there is room for improvements in both cases. The first configuration suffers from low throughput owing to the analysis of data from a single location. The second, on the other hand, is limited to relatively low frame rates and loses the benefit of single-photon-counting approaches. During the past few years, new developments in point-like and wide field detectors have started addressing some of these issues. Here, we describe our recent progresses towards increasing the throughput of single-molecule fluorescence spectroscopy in solution using parallel arrays of SPADs. We also discuss our development of large area photon-counting cameras achieving subnanosecond resolution for fluorescence lifetime imaging applications at the single-molecule level.

scholarly journals Single Molecule Blinking and Photobleaching Separated by Wide-Field Fluorescence Microscopy

2005 ◽  
Vol 109 (30) ◽  
pp. 6652-6658 ◽  
Author(s):  
Thomas Gensch ◽  
Martin Böhmer ◽  
Pedro F. Aramendía
Keyword(s):  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Wide Field

Styryl dye formation promoted by catalytic centers of piperazine bound to a silica surface traced by single molecule fluorescence microscopy

2017 ◽  
Vol 19 (31) ◽  
pp. 20984-20990 ◽  
Author(s):  
Aline M. Lino ◽  
Marcelo H. Gehlen
Keyword(s):  
Fluorescence Microscopy ◽  
Single Molecule ◽  
Silica Surface ◽  
Knoevenagel Condensation ◽  
Fluorescent Dye ◽  
New Method ◽  
Single Molecule Fluorescence ◽  
Single Molecule Level ◽  
Styryl Dye ◽  
Single Molecule Fluorescence Microscopy

Knoevenagel condensation producing a fluorescent dye is studied at a single molecule level establishing a new method to investigate the molecular rate of amino catalysis.

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