We use cookies to understand how you use our site and to improve your experience. This includes personalizing content and advertising. To learn more, click here. By continuing to use our site, you accept our use of cookies. Cookie Policy.

Features Partner Sites Information LinkXpress hp
Sign In
Advertise with Us

Download Mobile App




Raman Imaging Probes for Detecting Enzyme Activities Could Aid Disease Diagnosis

By LabMedica International staff writers
Posted on 02 Jun 2023

Enzymes play a crucial role in various biological processes, which makes them suitable biomarkers for disease detection. More...

For instance, cancer diagnosis often uses fluorescence imaging to identify cancer-related enzymes that have multiplied in cells affected by the disease. Given the heterogeneous nature of tumor tissues, being able to detect multiple enzyme activities at once could enhance the precision of cancer visualization and diagnosis. However, detecting multiple enzyme activities using fluorescence imaging can be challenging in heterogeneous tumor tissues and other complex biological phenomena. Raman spectral imaging, with its narrower spectral width, provides a promising alternative for multiplex biological imaging with molecular probes. Over time, a number of functional and activatable Raman probes (dyes) have been developed for bioanalyte detection, but their use for enzyme activity detection has been limited. In addition, prior design strategies have been unsuccessful in controlling the diffusion of enzyme-generated hydrolysis product of these probes, making it difficult to identify areas with targeted enzyme activity in tissues.

Now, a team of researchers led by Tokyo Institute of Technology (Tokyo Tech, Tokyo, Japan) has come up with a novel molecular design approach, taking cues from aggregation-based fluorescent probes. The team has developed activatable Raman probes based on 9CN-rhodol using a novel mechanism for Raman signal activation. The researchers have shown that Raman imaging has more potential for detecting multiple enzyme activities compared to fluorescence imaging. This innovative strategy allows the creation of highly activatable Raman probes that possess strong aggregation and multiplexing ability, providing a promising tool for expanding the range of Raman probes that can detect multiple enzyme activities in heterogeneous biological tissues.

The researchers first synthesized 9CN-rhodol derivatives and chose two, 9CN-JR and 9CN-JCR, to design the activatable Raman probes. They tested the enzyme detection capabilities of both probes in live cells, using a dual-color stimulated Raman scattering (SRS) imaging technique. Among the two, 9CN-JCR outperformed and proved to be the more effective probe for multiplexing. The team then isotope-labeled the nitril group of 9CN-JCR scaffold with Carbon-13 (13C) and Nitrogen-15 (15N), and developed two new isotope edited 9CN-JCR probes for γ-glutamyl transpeptidase and dipeptidyl peptidase-4 enzymes. These 9CN-JCR-based probes successfully detected all enzyme activities simultaneously in the live cell culture.

Additionally, the probes enabled ex vivo imaging of specific cell areas showing targeted enzyme activity in Drosophila wing disk and fat body. The high spatial selectivity and sensitivity exhibited by the 9CN-JCR probes were attributed to the electronic pre-resonance effect of the scaffold dye and aggregate formation of the hydrolysis products formed by probe-cell interaction. These rhodol-based probes could aggregate when reacting with enzymes, which enhanced their intracellular retention and boosted the SRS signal intensity during enzyme detection. Overall, the approach presented in this study could support the development of highly specific activatable Raman probes for simultaneous detection of multiple enzyme activities.

"Our aggregation-based molecular design strategy for Raman probes will offer substantial advantages for applications involving the investigation of enzyme activity associated with diseases and essential biological activities," said Professor Mako Kamiya of Tokyo Tech who led the research.

Related Links:
Tokyo Tech 


Gold Member
Quantitative POC Immunoassay Analyzer
EASY READER+
Collection and Transport System
PurSafe Plus®
Gold Member
Hematology Analyzer
Medonic M32B
Rapid Molecular Testing Device
FlashDetect Flash10
Read the full article by registering today, it's FREE! It's Free!
Register now for FREE to LabMedica.com and get access to news and events that shape the world of Clinical Laboratory Medicine.
  • Free digital version edition of LabMedica International sent by email on regular basis
  • Free print version of LabMedica International magazine (available only outside USA and Canada).
  • Free and unlimited access to back issues of LabMedica International in digital format
  • Free LabMedica International Newsletter sent every week containing the latest news
  • Free breaking news sent via email
  • Free access to Events Calendar
  • Free access to LinkXpress new product services
  • REGISTRATION IS FREE AND EASY!
Click here to Register








Channels

Hematology

view channel
Image: New evidence shows viscoelastic testing can improve assessment of blood clotting during postpartum hemorrhage (Photo courtesy of 123RF)

Viscoelastic Testing Could Improve Treatment of Maternal Hemorrhage

Postpartum hemorrhage, severe bleeding after childbirth, remains one of the leading causes of maternal mortality worldwide, yet many of these deaths are preventable. Standard care can be hindered by delays... Read more

Immunology

view channel
Image: The CloneSeq-SV approach can allow researchers to study how cells within high-grade serous ovarian cancer change over time (Photo courtesy of MSK)

Blood Test Tracks Treatment Resistance in High-Grade Serous Ovarian Cancer

High-grade serous ovarian cancer (HGSOC) is often diagnosed at an advanced stage because it spreads microscopically throughout the abdomen, and although initial surgery and chemotherapy can work, most... Read more

Industry

view channel
Image: The collaboration aims to improve access to Hb variant testing with the Gazelle POC diagnostic platform (Photo courtesy of Hemex Health)

Terumo BCT and Hemex Health Collaborate to Improve Access to Testing for Hemoglobin Disorders

Millions of people worldwide living with sickle cell disease and other hemoglobin disorders experience delayed diagnosis and limited access to effective care, particularly in regions where testing is scarce.... Read more
Copyright © 2000-2025 Globetech Media. All rights reserved.