Optimizing the Integration of Immunoreagents and Fluorescent Probes for Multiplexed High Content Screening Assays

2006 ◽  
pp. 189-194
Author(s):  
Kenneth A. Giuliano
BioTechniques ◽  
2021 ◽  
Vol 70 (6) ◽  
pp. 309-318
Author(s):  
Céline Rens ◽  
Tirosh Shapira ◽  
Sandra Peña-Diaz ◽  
Joseph D Chao ◽  
Tom Pfeifer ◽  
...  

Here the authors describe the development of AUTOptosis, an economical and rapid apoptosis monitoring method suitable for high-content and high-throughput screening assays. AUTOptosis is based on the quantification of nuclei intensity via staining with Hoechst 33342. First, the authors calibrated the method using standard apoptosis inducers in multiple cell lines. Next, the authors validated the applicability of this approach to high-content screening using a small library of compounds and compared it with the terminal deoxynucleotidyl transferase dUTP nick end labeling gold standard. Finally, the authors demonstrated the specificity of the method by using AUTOposis to detect apoptosis triggered by Mycobacterium tuberculosis intracellular infections.


2021 ◽  
Vol 28 ◽  
Author(s):  
Juan Cheng ◽  
Xin Li

: High-throughput screening facilitates the rapid identification of novel hit compounds; however, it remains challenging to design effective high-throughput assays, partially due to the difficulty of achieving sensitivity in the assay techniques. Among the various analytical methods that are used, fluorescence-based assays dominate owing to their high sensitivity and ease of operation. Recent advances in activity-based sensing/imaging have further expanded the availability of fluorescent probes as monitors for high-throughput screening of result outputs. In this study, we have reviewed various activity-based fluorescent probes used in high-throughput screening assays, emphasizing their structure-related working mechanisms. Moreover, we have explored the possibility of the development of additional and better probes to boost hit identification and drug development against various targets.


2016 ◽  
Vol 55 (52) ◽  
pp. 16026-16030 ◽  
Author(s):  
Pier Luca D'Alessandro ◽  
Nicole Buschmann ◽  
Markus Kaufmann ◽  
Pascal Furet ◽  
Frederic Baysang ◽  
...  

2016 ◽  
Vol 128 (52) ◽  
pp. 16260-16264 ◽  
Author(s):  
Pier Luca D'Alessandro ◽  
Nicole Buschmann ◽  
Markus Kaufmann ◽  
Pascal Furet ◽  
Frederic Baysang ◽  
...  

Author(s):  
Ann Cleary

Microinjection of fluorescent probes into living plant cells reveals new aspects of cell structure and function. Microtubules and actin filaments are dynamic components of the cytoskeleton and are involved in cell growth, division and intracellular transport. To date, cytoskeletal probes used in microinjection studies have included rhodamine-phalloidin for labelling actin filaments and fluorescently labelled animal tubulin for incorporation into microtubules. From a recent study of Tradescantia stamen hair cells it appears that actin may have a role in defining the plane of cell division. Unlike microtubules, actin is present in the cell cortex and delimits the division site throughout mitosis. Herein, I shall describe actin, its arrangement and putative role in cell plate placement, in another material, living cells of Tradescantia leaf epidermis.The epidermis is peeled from the abaxial surface of young leaves usually without disruption to cytoplasmic streaming or cell division. The peel is stuck to the base of a well slide using 0.1% polyethylenimine and bathed in a solution of 1% mannitol +/− 1 mM probenecid.


2001 ◽  
Vol 7 (S2) ◽  
pp. 1012-1013
Author(s):  
Uyen Tram ◽  
William Sullivan

Embryonic development is a dynamic event and is best studied in live animals in real time. Much of our knowledge of the early events of embryogenesis, however, comes from immunofluourescent analysis of fixed embryos. While these studies provide an enormous amount of information about the organization of different structures during development, they can give only a static glimpse of a very dynamic event. More recently real-time fluorescent studies of living embryos have become much more routine and have given new insights to how different structures and organelles (chromosomes, centrosomes, cytoskeleton, etc.) are coordinately regulated. This is in large part due to the development of commercially available fluorescent probes, GFP technology, and newly developed sensitive fluorescent microscopes. For example, live confocal fluorescent analysis proved essential in determining the primary defect in mutations that disrupt early nuclear divisions in Drosophila melanogaster. For organisms in which GPF transgenics is not available, fluorescent probes that label DNA, microtubules, and actin are available for microinjection.


2006 ◽  
Vol 82 (6) ◽  
pp. 1586 ◽  
Author(s):  
Prasun Mukherjee ◽  
Mintu Halder ◽  
Mark S. Hargrove ◽  
Jacob W. Petrich

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