Looking for breakthrough ideas for innovation challenges? Try Patsnap Eureka!

Application of near-infrared luminescent ruthenium complex to cell pH sensing

A dual-nuclear ruthenium complex, cell technology, applied in the field of cell pH sensing, can solve problems such as rare application reports

Inactive Publication Date: 2017-09-01
BEIJING NORMAL UNIVERSITY
View PDF6 Cites 8 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

However, there are very few reports on the application of such complexes in cellular pH sensing.

Method used

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
View more

Image

Smart Image Click on the blue labels to locate them in the text.
Viewing Examples
Smart Image
  • Application of near-infrared luminescent ruthenium complex to cell pH sensing
  • Application of near-infrared luminescent ruthenium complex to cell pH sensing
  • Application of near-infrared luminescent ruthenium complex to cell pH sensing

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0016] Embodiment 1: cell culture and cytotoxicity experiment

[0017] Cervical cancer HeLa cells were cultured with cell culture medium at 370°C in CO 2 cultured in an incubator (95% relative humidity, 5% CO 2 ). The in vitro cytotoxicity of the complex was determined on a high-content analysis system produced by Molecular Devices. Take cervical cancer HeLa cells that are in the exponential growth phase, make cell suspension with culture medium, and count with a cell counting plate, according to 6×10 3 Cells / well were seeded in 96-well plates for subculture. After 24 hours of cell subculture, add the dimethyl sulfoxide solution of the complex to make the final concentration 10-80 μM. 100 μL of medium and cisplatin were used as negative and positive controls, respectively. The 96-well plate was placed in an incubator with 95% relative humidity and 5% carbon dioxide at 37°C for 48 hours. Then stained with 10 μg / mL Hoechst 33342 and 1 μg / mL propidium iodide PI in phosphate b...

Embodiment 2

[0019] Example 2: Flow Cytometry Analysis

[0020] Cervical cancer HeLa cells were digested with trypsin, suspended in 5 mL of phosphate buffer, and centrifuged at 1000 rpm for 10 minutes. Discard the supernatant and add 1 mL of phosphate buffer to resuspend the cells. A sample containing 20000 cells was analyzed by a NovoCyte type flow cytometer (ACEA, USA). The excitation wavelength is 488nm, and the emitted photons are collected through a 780 / 20nm filter.

[0021] We explored the optimal concentration of the complexes and the optimal incubation time by flow cytometry. It can be seen from Figure 3 that when the concentration of the complex is 80 μM, the luminous intensity of the cells is still not saturated. Due to the limitation of the solubility of the complex, we choose a concentration of 40 μM complex close to the maximum luminous intensity of the cells in our experiment. It can be seen from Figure 4 that when the incubation time reaches 8 hours, the cells reach the m...

Embodiment 3

[0022]Example 3: HeLa cell pH imaging

[0023] In order to complete laser confocal imaging, cervical cancer HeLa in 20mm 2 Cells were cultured in laser confocal microscope dishes (1.0 × 10 per well 4 cells). Cells at 37°C in 5% CO 2 The atmosphere was maintained for 24 hours until 70% confluency was achieved. The 10 mM complex dimethyl sulfoxide solution was diluted with cell culture medium to a final concentration of 40 μM. After the cells were incubated for 7.5 hours, the excess complexes were washed 3 times with phosphate buffer (pH 7.4) and further treated for 10 minutes. Then, the cells were washed three times with phosphate buffer (pH 7.4), and incubated with 10 μM nigericin at different pH values ​​(4.0, 4.5, 5.0, 5.5, 6.0, 7.0 and 7.4) for 15 minutes. Zeiss AXIO Observer A1 ordinary microscope at 37°C for luminescent imaging (excitation wavelength 450-490nm, collection of emitted photons with emission wavelength greater than 515nm). Image analysis was performed w...

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

PUM

No PUM Login to View More

Abstract

The invention discloses application of a near-infrared luminescent ruthenium complex to cell staining and pH sensing in a probe cell. The complex has almost no cytotoxicity at the staining concentration, and can stain a cell nucleus and cytoplasm. Through change of near-infrared luminescence intensity of the cell along with pH, the pH in the cell can be detected by a flow cytometry and a microscopic imaging technology.

Description

technical field [0001] The invention relates to the field of cell pH sensing, in particular to the application of a binuclear ruthenium complex in the pH measurement of tumor cells. Background technique [0002] Cell pH is closely related to cell function [R.A.Gottlieb, J.Nordberg, E.Skowronski, B.M.Babior, Apoptosis induced in Jurkat cells by several agents is preceded by intracellular acidification, Proc.Natl.Acad.Sci.USA, 1996,93,654. ]. Generally, there are two wide pH ranges in cells, such as the pH ranges of cytoplasm and lysosomes are 6.8-7.4 and 4.5-6.0 respectively [F.Galindo, M.I.Burguete, L.Vigara, S.V.Luis, N.Kabir , J. Gavrilovic, D.A. Russell, Synthetic macrocyclic peptidomimetics as tunable pH probes for the fluorescence imaging of acidic organelles in live cells, Angew. Chem. Int. Ed. Engl., 2005, 44, 6504.]. Cellular dysfunction, diseases such as cancer and Alzheimer's disease can cause the intracellular pH value to deviate from the normal value [H.Izumi, ...

Claims

the structure of the environmentally friendly knitted fabric provided by the present invention; figure 2 Flow chart of the yarn wrapping machine for environmentally friendly knitted fabrics and storage devices; image 3 Is the parameter map of the yarn covering machine
Login to View More

Application Information

Patent Timeline
no application Login to View More
Patent Type & Authority Applications(China)
IPC IPC(8): C07F15/00C09K11/06G01N15/14G01N21/63
CPCC07F15/0053C09K11/06C09K2211/185G01N15/1434G01N21/63
Inventor 王克志孟婷婷刘进
Owner BEIJING NORMAL UNIVERSITY
Who we serve
  • R&D Engineer
  • R&D Manager
  • IP Professional
Why Patsnap Eureka
  • Industry Leading Data Capabilities
  • Powerful AI technology
  • Patent DNA Extraction
Social media
Patsnap Eureka Blog
Learn More
PatSnap group products

Browse by: Latest US Patents, China's latest patents, Technical Efficacy Thesaurus, Application Domain, Technology Topic, Popular Technical Reports.

© 2024 PatSnap. All rights reserved.Legal|Privacy policy|Modern Slavery Act Transparency Statement|Sitemap|About US| Contact US: help@patsnap.com