scholarly journals A far-red fluorescent chemogenetic reporter for in vivo molecular imaging

2020 ◽  
Author(s):  
Chenge Li ◽  
Alison G. Tebo ◽  
Marion Thauvin ◽  
Marie-Aude Plamont ◽  
Michel Volovitch ◽  
...  
Keyword(s):  
Protein Interactions ◽  
Zebrafish Embryo ◽  
Tissue Imaging ◽  
Live Cells ◽  
Protein Protein Interactions ◽  
Deep Tissue ◽  
Red Fluorescence ◽  
Genetic Encoding ◽  
Deep Tissue Imaging

AbstractFar-red emitting fluorescent labels are highly desirable for spectral multiplexing and deep tissue imaging. Here, we describe the generation of frFAST (far-red Fluorescence Activating and absorption Shifting Tag), a 14-kDa monomeric protein that forms a bright far-red fluorescent assembly with (4-hydroxy-3-methoxy-phenyl)allylidene rhodanine (HPAR-3OM). As HPAR-3OM is essentially non-fluorescent in solution and in cells, frFAST can be imaged with high contrast in presence of free HPAR-3OM, which allowed the rapid and efficient imaging of frFAST fusions in live cells, zebrafish embryo/larvae and chicken embryo. Beyond enabling genetic encoding of far-red fluorescence, frFAST allowed the design of a far-red chemogenetic reporter of protein-protein interactions, demonstrating its great potential for the design of innovative far-red emitting biosensors.

The one-pot synthesis of core/shell/shell CdTe/CdSe/ZnSe quantum dots in aqueous media for in vivo deep tissue imaging

2011 ◽  
Vol 21 (9) ◽  
pp. 2877 ◽  
Author(s):  
Shohei Taniguchi ◽  
Mark Green ◽  
Sarwat B. Rizvi ◽  
Alexander Seifalian
Keyword(s):  
Quantum Dots ◽  
Aqueous Media ◽  
Tissue Imaging ◽  
Core Shell ◽  
Deep Tissue ◽  
One Pot ◽  
One Pot Synthesis ◽  
Deep Tissue Imaging ◽  
The One

scholarly journals Assay Systems for Profiling Deubiquitinating Activity

2020 ◽  
Vol 21 (16) ◽  
pp. 5638
Author(s):  
Jinhong Cho ◽  
Jinyoung Park ◽  
Eunice EunKyeong Kim ◽  
Eun Joo Song
Keyword(s):  
Protein Degradation ◽  
Protein Interactions ◽  
Live Cells ◽  
Deubiquitinating Enzyme ◽  
Protein Protein Interactions ◽  
Deubiquitinating Enzymes ◽  
Cell Lysates ◽  
Cellular Processes

Deubiquitinating enzymes regulate various cellular processes, particularly protein degradation, localization, and protein–protein interactions. The dysregulation of deubiquitinating enzyme (DUB) activity has been linked to several diseases; however, the function of many DUBs has not been identified. Therefore, the development of methods to assess DUB activity is important to identify novel DUBs, characterize DUB selectivity, and profile dynamic DUB substrates. Here, we review various methods of evaluating DUB activity using cell lysates or purified DUBs, as well as the types of probes used in these methods. In addition, we introduce some techniques that can deliver DUB probes into the cells and cell-permeable activity-based probes to directly visualize and quantify DUB activity in live cells. This review could contribute to the development of DUB inhibitors by providing important information on the characteristics and applications of various probes used to evaluate and detect DUB activity in vitro and in vivo.

scholarly journals Ultrabright red AIEgens for two-photon vascular imaging with high resolution and deep penetration

2018 ◽  
Vol 9 (10) ◽  
pp. 2705-2710 ◽  
Author(s):  
Wei Qin ◽  
Pengfei Zhang ◽  
Hui Li ◽  
Jacky W. Y. Lam ◽  
Yuanjing Cai ◽  
...  
Keyword(s):  
High Resolution ◽  
Vascular Imaging ◽  
Tissue Imaging ◽  
Deep Penetration ◽  
High Contrast ◽  
Deep Tissue ◽  
Two Photon ◽  
High Penetration ◽  
Deep Tissue Imaging

A successful strategy for the design of ultrabright red luminogens with aggregation-induced emission (AIE) features is reported. The AIE dots can be utilized as efficient fluorescent probes for in vivo deep-tissue imaging with high penetration depth and high contrast.

scholarly journals Label-free microendoscopy using a micro-needle imaging probe for in vivo deep tissue imaging

2020 ◽  
Vol 11 (9) ◽  
pp. 4976
Author(s):  
Kwanjun Park ◽  
June Hoan Kim ◽  
Taedong Kong ◽  
Woong Sun ◽  
Jonghwan Lee ◽  
...  
Keyword(s):  
Tissue Imaging ◽  
Label Free ◽  
Deep Tissue ◽  
Imaging Probe ◽  
Deep Tissue Imaging

Five-nanometer ZnSn2O4:Cr,Eu ultra-small nanoparticles as new near infrared-emitting persistent luminescent nanoprobes for cellular and deep tissue imaging at 800 nm

Nanoscale ◽  
2017 ◽  
Vol 9 (25) ◽  
pp. 8631-8638 ◽  
Author(s):  
Jin-Lei Li ◽  
Jun-Peng Shi ◽  
Cheng-Cheng Wang ◽  
Peng-Hui Li ◽  
Zhen-Feng Yu ◽  
...  
Keyword(s):  
Near Infrared ◽  
Tissue Imaging ◽  
Deep Tissue ◽  
Deep Tissue Imaging ◽  
Small Nanoparticles

Schematic illustration of the synthesis, functionalization and repeated in vivo simulated deep tissue imaging of ZSO NPLNPs.

In vivo deep tissue imaging with long wavelength multiphoton excitation

2010 ◽  
Author(s):  
Demirhan Kobat ◽  
Michael E. Durst ◽  
Nozomi Nishimura ◽  
Angela W. Wong ◽  
Chris B. Schaffer ◽  
...  
Keyword(s):  
Tissue Imaging ◽  
Multiphoton Excitation ◽  
Deep Tissue ◽  
Long Wavelength ◽  
Deep Tissue Imaging

scholarly journals Oligomerization of the UDP-glucuronosyltransferase 1A Proteins

2006 ◽  
Vol 282 (7) ◽  
pp. 4821-4829 ◽  
Author(s):  
Theresa N. Operaña ◽  
Robert H. Tukey
Keyword(s):  
Protein Interactions ◽  
Fluorescent Protein ◽  
Resonance Energy Transfer ◽  
Resonance Energy ◽  
Conclusive Evidence ◽  
Live Cells ◽  
Protein Protein Interactions ◽  
Dietary Nutrients ◽  
Membrane Bound

UDP-glucuronosyltransferases (UGTs) are membrane-bound proteins localized to the endoplasmic reticulum and catalyze the formation of β-d-glucopyranosiduronic acids (glucuronides) using UDP-glucuronic acid and acceptor substrates such as drugs, steroids, bile acids, xenobiotics, and dietary nutrients. Recent biochemical evidence indicates that the UGT proteins may oligomerize in the membrane, but conclusive evidence is still lacking. In the present study, we have used fluorescence resonance energy transfer (FRET) to study UGT1A oligomerization in live cells. This technique demonstrated that UGT1A1, UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, UGT1A9, and UGT1A10 self-oligomerize (homodimerize). Heterodimer interactions were also explored, and it was determined that UGT1A1 was capable of binding with UGT1A3, UGT1A4, UGT1A6, UGT1A7, UGT1A8, UGT1A9, and UGT1A10. In addition to the in vivo FRET analysis, UGT1A protein-protein interactions were demonstrated through co-immunoprecipitation experiments. Co-expression of hemagglutinin-tagged and cyan fluorescent protein-tagged UGT1A proteins, followed by immunoprecipitation with anti-hemagglutinin beads, illustrated the potential of each UGT1A protein to homodimerize. Co-immunoprecipitation results also confirmed that UGT1A1 was capable of forming heterodimer complexes with all of the UGT1A proteins, corroborating the FRET results in live cells. These preliminary studies suggest that the UGT1A family of proteins form oligomerized complexes in the membrane, a property that may influence function and substrate selectivity.

3D printed optical phantoms and deep tissue imaging for in vivo applications including oral surgery

2017 ◽  
Author(s):  
Brian Z. Bentz ◽  
Alfonso Costas ◽  
Vaibhav Gaind ◽  
Jose M. Garcia ◽  
Kevin J. Webb
Keyword(s):  
Oral Surgery ◽  
Tissue Imaging ◽  
Deep Tissue ◽  
Deep Tissue Imaging ◽  
3D Printed

Ablation of Red Stable Transfected Prostate Cancer Cell Lines by C-CPE Gold-Nanoparticle Mediated Laser Intervention

2021 ◽  
Author(s):  
Suhayla Alnajjar ◽  
Ingo Nolte ◽  
Annegret Becker ◽  
Tina Kostka ◽  
Jan Torben Schille ◽  
...  
Keyword(s):  
Prostate Cancer ◽  
Gold Nanoparticle ◽  
Cell Lines ◽  
Cancer Cell ◽  
Tissue Imaging ◽  
Deep Tissue ◽  
Deep Tissue Imaging

Abstract Background: Claudin (CLDN) proteins have been described to be found and accordingly targeted to evaluate novel therapeutic approaches. C-terminus of Clostridium perfringens enterotoxin (C-CPE) binds efficiently several claudins and thus recombinant C-CPE conjugated to gold nanoparticles (AuNPs) has been used for cancer cell targeting using gold nanoparticle- mediated laser perforation (GNOME-LP). Cancer cells inoculation is routinely used to generate in vivo models to evaluate novel therapeutic approaches in prostate cancer. However, detailed characterization of cancer spreading and early tumor development and therapeutic response is often limited as conventional cell lines do not allow advanced deep tissue imaging.Methods: two canine prostate cancer cell lines were stably transfected with red fluorescent protein (RFP), followed by G418 selection. RFP marker as well as CLDN3, -4 and -7 expression was comparatively confirmed by flow cytometry, qPCR and immunofluorescences. For cancer cells targeting, GNOME-LP at a laser fluence of 72 mJ/cm² and a scanning speed of 0.5 cm/s was used. Statistical analysis was performed using SAS software 7.1, Dunnett´s Multiple Comparison Test and Student´s two-sided t-test. Differences were considered statistically significant for p<0.05.Results: we established two canine prostate carcinoma cell lines, stably expressing RFP allowing perspective deep tissue imaging. Directed C-CPE-AuNP binding to native and RFP transfected cells verified the capability to specifically target CLDN receptors. Cancer cell ablation was demonstrated in vitro setting using a combination of gold nanoparticle mediated laser perforation and C-CPE-AuNPs treatment reducing tumor cell viability to less than 10 % depending on cell line. Conclusion: the results confirm that this therapeutic approach can be used efficiently to target prostate carcinoma cells carrying a marker protein allowing deep tissue imaging. The established cell lines and the verified proof of concept in vitro study provide the basis for perspective Xenograft model in vivo studies. The introduce red fluorescence enables deep tissue imaging in living animals and therefore detailed characterization of tumor growth and subsequently possible tumor ablation through C-CPE-AuNPs treatment.

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