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

Biosensors, and method and kits for using same

Inactive Publication Date: 2006-07-06
BELLEMARE FRANCOIS +2
View PDF2 Cites 6 Cited by
  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Benefits of technology

[0010] There is also provided a method for detecting toxic molecules comprising the use of these biosensors in combination with a fluorometer advantageously permitting the use of microprocessor since the measurement is made with a non-modulated light rather than pulsed light. The pulsed light permits the measurement of photochemical and non photochemical parameters in the whole plants or algaes. In isolated thylakoid membranes, because certain non photochemical parameters can not be obtained, pulsed light may not be necessary. Furthermore, pulsed light fluorometers are relatively complex, costly and bulky apparatuses. Fluorometers using non-modulated light do not have these drawbacks and are nevertheless able to adequately measure the photosynthetic efficiency of thylakoid membranes.

Problems solved by technology

Furthermore, pulsed light fluorometers are relatively complex, costly and bulky apparatuses.
Fluorometers using non-modulated light do not have these drawbacks and are nevertheless able to adequately measure the photosynthetic efficiency of thylakoid membranes.

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
  • Biosensors, and method and kits for using same
  • Biosensors, and method and kits for using same
  • Biosensors, and method and kits for using same

Examples

Experimental program
Comparison scheme
Effect test

example 1

Fluorometer

[0038] In a specific embodiment, a fluorometer continuously illuminates weakly the photosynthetic biological material with a light emitting diode (LED) emitting light at a wavelength of 475 or at 660 nm. Better results were obtained with the 475 nm wavelength. In order to obtain a higher sensitivity for measuring the photosynthetic response to toxic molecules, the intensity of the low excitation light was adjusted so as to obtain a F1 fluorescence level slightly higher than F0.

[0039] Fluorescence measured 2 sec after turning on one LED is F1. Three additional LEDs are then turned on so as to induce actinic illumination F2. Dispersion of LEDs is illustrated in FIG. 1. Light beams are oriented so as to form together a point of convergence precisely at the level where the photodiode is read. After a specific time, selected according to the nature of the thylakoid membranes used (eg. source of thylakoids, etc.) (Table 1), of the background noise and of the level of sensitiv...

example 2

Preparation of Thylakoid Membranes

[0043] All steps are conducted in the dark or under green light and at cold temperature (samples on ice or procedure in cold room). Thylakoid membranes are isolated from 100 g of deveined spinach leaves. Spinach leaves are then crushed in a mixer with homogenising a buffer comprising TES-NaOH 20 mM pH 7.5, sorbitol 330 mM and MgCl2 5 mM. Other conventional buffers could be used including TES, Hepes, Tris, phosphate, tricine and MOPs. The homogenate is filtered on a cheese cloth and the filtrate is centrifuged 2 min at 2500×g at 40° C. on a Eppendorf™5810-R, rotor #A-4-44.

[0044] The pellet is then resuspended in a hypotonic solution consisting in 1 in 20 of the homogenising buffer. This step is used to lyse the chloroplast membranes. The resulting solution is then crushed in a Wheaton™ mixer and centrifuged 3 min. at 3500×. The resulting pellet contains the purified membranes. They are then resuspended in a buffer consisting of TES-NaOH 20 mM pH 7....

example 3

Preparation of Ready to Use Stabilised Thylakoid Membranes Formulation

[0045] A preparation of 2 to 3 mg / mL of thylakoid membranes were mixed with a pH 6.5 phosphate buffer containing: 0.02 M phosphate, sucrose 300 mM, NH4Cl 10 mM, MgCl2 20 mM, EDTA 10 mM, polyvinylpyrrolidine (PVP) (125 μL of solution 20% for 1 mL) and liposomes constituted of phosphatidylcholine and phosphatidylglycerol in a ratio of 10:1 to obtain a solution of 0.0125 mg of liposome / mL of chlorophyll / thylakoid membranes buffered solution. In this stabilised thylakoid membrane formulation, the final concentrations of PVP is of 2% v / v, the final concentration of liposomes is of 0.0125 mg / mL v / v and that of chlorophyll is of 0.125mg / mL v / v.

[0046] The PVP increased the stability of the thylakoid preparations as compared to the same preparation without PVP but a useful stability was nevertheless obtained even without PVP. The amount of PVP may be varied without affecting the stability of the thylakoid membranes betwe...

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

A stabilised thylakoid membrane formulation comprising thylakoid membranes in a buffered solution and liposomes, wherein the formulation has a ratio of chlorophyll / liposomes of at least about 10:1. The invention further comprises a method for detecting or quantifying the presence of toxic molecules in a fluid sample, comprising obtaining a stabilised thylakoids membranes formulation; and assessing the photosynthetic efficiency of the thylakoid membranes formulation in the presence of said sample, whereby the molecules are detected when the photosynthetic efficiency is measurably different in the presence versus in the absence of the sample. The present invention also comprises kits using the stabilised thylakoid membranes formulation for detecting and / or quantifying the toxic molecule in a fluid sample.

Description

FIELD OF THE INVENTION [0001] The present invention relates to biosensors, and method and kits for using same. More specifically, the present invention is concerned with photosynthesis-based biosensors, and method and kits for using same. BACKGROUND OF THE INVENTION [0002] Many assays for detecting toxic molecules in effluents are inefficient when these molecules are present at very low concentration. Assays that are efficient for this purpose such as those made with living organisms and cells like algaes, daphneas, trouts, hepatocytes, or worms are time consuming (up to 72 hours). These methods investigate the presence of toxic molecules in water by measuring the survival of these organisms and cells in contact of the water being tested. [0003] Assays using the response of photosynthetic organs to toxic molecules have been recently devised which in addition to reducing the response time of the assay to 10 minutes are advantageously sensitive to molecules affecting photosynthetic el...

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
IPC IPC(8): G01N31/22G01N33/50
CPCG01N33/5097G01N2430/20Y10T436/173076Y10T436/141111Y10T436/201666Y10T436/206664Y10T436/175383Y10T436/16
Inventor BELLEMARE, FRANCOISBOUCHER, NATHALIELORRAIN, LUCIE
Owner BELLEMARE FRANCOIS
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