A Quantitative Measurement Method of FRET Efficiency Applied to Cell Apoptosis Detection

A technique for quantitative measurement and apoptosis, applied in material excitation analysis, fluorescence/phosphorescence, etc., can solve the problems of cumbersome steps and high equipment cost

Inactive Publication Date: 2021-06-29
FUJIAN NORMAL UNIV
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  • Abstract
  • Description
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Problems solved by technology

However, when used to analyze the process of apoptosis, the steps of these methods are too cumbersome, and the cost of equipment required is relatively high.

Method used

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  • A Quantitative Measurement Method of FRET Efficiency Applied to Cell Apoptosis Detection
  • A Quantitative Measurement Method of FRET Efficiency Applied to Cell Apoptosis Detection
  • A Quantitative Measurement Method of FRET Efficiency Applied to Cell Apoptosis Detection

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Embodiment Construction

[0031] The invention will be further described below with reference to the accompanying drawings and examples.

[0032] Please refer to figure 1 In this embodiment, in order to better test the effectiveness of the measurement method, a binding, a receptor concentration ratio 1: 1 plasmid C32V simulated apoptosis detection FRET probe, and measuring the FRET apparent efficiency of the plasmid;

[0033] 1, to be tested simulation probe

[0034] C32V is supplied to the surcharge 1: 1

[0035] 2, refer to the serial plasmid

[0036] VCV CERULEAN - VENUS series particles supply receptor concentration ratio 1: 2, FRET apparent efficiency E app串 = 69.3 ± 1.0% (measured by FLIM method)

[0037] 3, measurement instrument and filter set

[0038] Microscope: Olympus IX73

[0039] CCD: Hamamatsu) Orca-Flash 4.0 LT C1140-42U CMOS

[0040] Excited light source: metal halogen lamp Olympus u-hglgps

[0041] Objective lens: Olympus 40X / NA 1.3 OIL

[0042] Different filter set (Chroma): excitation...

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Abstract

The invention relates to a method for quantitatively measuring FRET efficiency applied to cell apoptosis detection. The FRET probe for cell apoptosis detection of the present invention has the characteristics of a concentration ratio of 1:1 between the donor fluorescent group and the acceptor fluorescent group, and does not need to correct the fluorescent crosstalk of the system, nor does it need to measure the sensitization and quenching conversion factor of the system, Only one tandem plasmid with known parameters is needed as a reference, which can quickly realize the quantitative monitoring of the spatiotemporal process of apoptosis. The invention has low requirement on instrument configuration and low cost, and has the advantages of being able to realize dynamic and quantitative measurement of FRET apparent efficiency and the like.

Description

Technical field [0001] The present invention relates to a quantitative measurement method of an apparent efficiency of fluorescence resonance energy transfer (FRET), and specifically, an FRET efficiency quantitative measurement method applied to apoptosis detection. Background technique [0002] Fluorescent resonance energy transfer (FRET) technology has a wide range of applications in biophysics and biochemistry. Combined with fluorescence microscopy can accurately locate biomolecules and perform real-time monitoring of molecular dynamics, interaction, protein-DNA interaction, protease activity, protein conformation, etc. Changes. Equisical efficiency change. [0003] The rapid and accurate detection of apoptosis is important in the field of biological and medical, and the existing study refers to the key role in caspases in the process of apoptosis. At present, the FRET probes for apoptosis detection are mainly studied by detecting Caspase-2, Caspase-3, Caspase-8, Caspase-9 cys...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): G01N21/64
Inventor 吴志伟林居强黄义梅谢树森
Owner FUJIAN NORMAL UNIV
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