Fluorescent Labeling Techniques for Investigation of Fibronectin Fibrillogenesis (Labeling Fibronectin Fibrillogenesis)

2009 ◽  
pp. 261-274 ◽  
Author(s):  
Roumen Pankov ◽  
Albena Momchilova
Keyword(s):  
Fluorescent Labeling

Rhodanol synthons for fluorescent labeling and drug conjugates

1996 ◽  
Vol 61 (s1) ◽  
pp. 290-293 ◽  
Author(s):  
Maria Spassova ◽  
Pavol Kois ◽  
Kyoichi Watanabe
Keyword(s):  
Fluorescent Labeling ◽  
Drug Conjugates

027 GABAergic Neurons in the Dorsal Raphe Nucleus Are under the Influence of GABAergic Inputs from the Nucleus Pontis Oralis

SLEEP ◽  
2021 ◽  
Vol 44 (Supplement_2) ◽  
pp. A12-A12
Author(s):  
Jianhua Zhang ◽  
Mingchu Xi ◽  
Simon Fung ◽  
Charles Tobin ◽  
Sharon Sampogna ◽  
...  
Keyword(s):  
Dorsal Raphe Nucleus ◽  
Dorsal Raphe ◽  
Gabaergic Neurons ◽  
Fluorescent Labeling ◽  
Raphe Nucleus ◽  
Gaba A ◽  
Adult Cats ◽  
Immunohistochemical Techniques ◽  
Fluorescence Immunohistochemistry ◽  
Dorsal Raphé

Abstract Introduction Our previous study has shown that there is a direct connection between GABAergic neurons in the nucleus pontis oralis (NPO) and neurons of the dorsal raphe nucleus (DR), providing a morphological basis for the hypothesis that GABAergic inhibitory processes in NPO play an important role in the generation and maintenance of wakefulness as well as active (REM) sleep through the interaction with neurons in the DR. However, the target of such a GABAergic projection from the NPO within the DR is unknown. In the present study, a double-fluorescent labeling technique was employed to examine the target of GABAergic inputs to the DR. Methods Adult cats were deeply anesthetized and perfused transcardially. Subsequently, the brainstem containing the DR was removed, postfixed and cut into 15 μm coronal sections with a Reichert-Jung cryostat. The sections were immunostained with antibodies against GABA-A or GABA-B receptors and GABA following the procedure of double fluorescence immunohistochemistry. Results Under fluorescence microscopy, a large number of neurons were labeled with antibodies against either GABA-A receptor or GABA-B receptor. In addition, neurons labeled with antibody against GABA were observed in the DR. With double fluorescence immunohistochemical techniques, some neurons labeled by anti-GABA antibody were also stained with antibodies against GABA-A or GABA-B receptors. Conclusion The expression of GABA-A or GABA-B receptors by GABAergic neurons in the DR indicates that GABAergic neurons in the DR receive GABAergic inputs. Our previous study has demonstrated that these GABAergic inputs are from the NPO. These data provide a morphological foundation to support our hypothesis that, during wakefulness, NPO GABAergic “Executive” neurons suppress “Second-Order” GABAergic neurons in the DR, which, in turn, activate (disinhibit) serotonergic wake-on neurons in this nucleus. Support (if any) NS092383

scholarly journals Split-wrmScarlet and split-sfGFP: Tools for faster, easier fluorescent l abeling of endogenous proteins in Caenorhabditis elegans

Genetics ◽  
2021 ◽  
Author(s):  
Jérôme Goudeau ◽  
Catherine S Sharp ◽  
Jonathan Paw ◽  
Laura Savy ◽  
Manuel D Leonetti ◽  
...  
Keyword(s):  
Fluorescent Protein ◽  
Fluorescent Proteins ◽  
Fluorescent Labeling ◽  
Large Fragment ◽  
C Elegans ◽  
High Affinity ◽  
Red Fluorescent Proteins ◽  
Invaluable Tool ◽  
Endogenous Proteins ◽  
Fluorescent Tags

Abstract We create and share a new red fluorophore, along with a set of strains, reagents and protocols, to make it faster and easier to label endogenous C. elegans proteins with fluorescent tags. CRISPR-mediated fluorescent labeling of C. elegans proteins is an invaluable tool, but it is much more difficult to insert fluorophore-size DNA segments than it is to make small gene edits. In principle, high-affinity asymmetrically split fluorescent proteins solve this problem in C. elegans: the small fragment can quickly and easily be fused to almost any protein of interest, and can be detected wherever the large fragment is expressed and complemented. However, there is currently only one available strain stably expressing the large fragment of a split fluorescent protein, restricting this solution to a single tissue (the germline) in the highly autofluorescent green channel. No available C. elegans lines express unbound large fragments of split red fluorescent proteins, and even state-of-the-art split red fluorescent proteins are dim compared to the canonical split-sfGFP protein. In this study, we engineer a bright, high-affinity new split red fluorophore, split-wrmScarlet. We generate transgenic C. elegans lines to allow easy single-color labeling in muscle or germline cells and dual-color labeling in somatic cells. We also describe a novel expression strategy for the germline, where traditional expression strategies struggle. We validate these strains by targeting split-wrmScarlet to several genes whose products label distinct organelles, and we provide a protocol for easy, cloning-free CRISPR/Cas9 editing. As the collection of split-FP strains for labeling in different tissues or organelles expands, we will post updates at doi.org/10.5281/zenodo.3993663

scholarly journals Gezielte Zellsortierung in der Einzelzellgenomik

BIOspektrum ◽  
2021 ◽  
Vol 27 (3) ◽  
pp. 274-276
Author(s):  
Morgan S. Sobol ◽  
Anne-Kristin Kaster
Keyword(s):  
Dark Matter ◽  
Single Cell ◽  
Cell Sorting ◽  
Genetic Information ◽  
Fluorescent Labeling ◽  
Single Cell Genomics ◽  
Rare Biosphere ◽  
Prokaryotic Genomes ◽  
Selection Of

AbstractSingle cell genomics (SCG) can provide reliable context for assembled genome fragments on the level of individual prokaryotic genomes and has rapidly emerged as an essential complement to cultivation-based and metagenomics research approaches. Targeted cell sorting approaches, which enable the selection of specific taxa by fluorescent labeling, compatible with subsequent single cell genomics offers an opportunity to access genetic information from rare biosphere members which would have otherwise stayed hidden as microbial dark matter.

Fluorescent labeling of s2T-incorporated DNA and m5s2U-modified RNA

2021 ◽  
pp. 1-13
Author(s):  
Ping Yu ◽  
Honglin Zhou ◽  
Yuanyuan Li ◽  
Zhifeng Du ◽  
Rui Wang
Keyword(s):  
Fluorescent Labeling

scholarly journals Characterization of Liposomes Using Quantitative Phase Microscopy (QPM)

Pharmaceutics ◽  
2021 ◽  
Vol 13 (5) ◽  
pp. 590
Author(s):  
Jennifer Cauzzo ◽  
Nikhil Jayakumar ◽  
Balpreet Singh Ahluwalia ◽  
Azeem Ahmad ◽  
Nataša Škalko-Basnet
Keyword(s):  
Rapid Development ◽  
Fluorescent Labeling ◽  
Label Free ◽  
Batch Mode ◽  
Full Field ◽  
Quantitative Phase ◽  
Phase Microscopy ◽  
Quantitative Phase Microscopy ◽  
Tracking Ability

The rapid development of nanomedicine and drug delivery systems calls for new and effective characterization techniques that can accurately characterize both the properties and the behavior of nanosystems. Standard methods such as dynamic light scattering (DLS) and fluorescent-based assays present challenges in terms of system’s instability, machine sensitivity, and loss of tracking ability, among others. In this study, we explore some of the downsides of batch-mode analyses and fluorescent labeling, while introducing quantitative phase microscopy (QPM) as a label-free complimentary characterization technique. Liposomes were used as a model nanocarrier for their therapeutic relevance and structural versatility. A successful immobilization of liposomes in a non-dried setup allowed for static imaging conditions in an off-axis phase microscope. Image reconstruction was then performed with a phase-shifting algorithm providing high spatial resolution. Our results show the potential of QPM to localize subdiffraction-limited liposomes, estimate their size, and track their integrity over time. Moreover, QPM full-field-of-view images enable the estimation of a single-particle-based size distribution, providing an alternative to the batch mode approach. QPM thus overcomes some of the drawbacks of the conventional methods, serving as a relevant complimentary technique in the characterization of nanosystems.

scholarly journals Fluorescent labeling of plant chromosomes in suspension by FISH

2005 ◽  
Vol 80 (1) ◽  
pp. 35-39 ◽  
Author(s):  
Youzhi Ma ◽  
Jai-Heon Lee ◽  
Lian Cheng Li ◽  
Susumu Uchiyama ◽  
Nobuko Ohmido ◽  
...  
Keyword(s):  
Fluorescent Labeling ◽  
Plant Chromosomes

Plate-based biochemical assay using quantum dots as a fluorescent labeling agent

2005 ◽  
Vol 108 (1-2) ◽  
pp. 713-720 ◽  
Author(s):  
Chin-Ping Huang ◽  
Hong-Wei Liu ◽  
Chao-Yun Tsao ◽  
Li-Te Yin ◽  
Su-Feng Chiu ◽  
...  
Keyword(s):  
Quantum Dots ◽  
Fluorescent Labeling ◽  
Biochemical Assay
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