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Viable cell analysis system for allergen screening

A technology of live cells and allergens, applied in the field of live cell analysis of amines, can solve the problem of many uncontrollable factors

Inactive Publication Date: 2014-12-24
TIANJIN MOSBIO SCI & TECH CO LTD
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

The experiment involves three steps: cell preparation, histamine release and detection, and there are many uncontrollable factors

Method used

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  • Viable cell analysis system for allergen screening

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Experimental program
Comparison scheme
Effect test

Embodiment approach 1

[0022] Embodiment 1: Comparison of the number of cells retained on the filter membrane and the amount of histamine released

[0023] After density gradient centrifugation to prepare mast cells, the cells were counted, and the microchips were incubated at 37°C and filled with saline solution. Several microliters of cell suspensions of different concentrations are injected using a micropipette through a connecting tube, which is connected to a syringe pump using a capillary, and the buffered injection injects the cells into the effector cell compartment. In order to detect whether the cells are leaking, after the cells are introduced, toluidine blue is injected, and the chamber of the histamine reaction pool is observed using a microscope. In addition, collect the eluate at a flow rate of 10 μl / min for 10 minutes, and stain with toluidine blue to check for cell leakage. Introduce fluorescent derivative solution (containing 0.5% o-phthalaldehyde, 1% methanol and 0.6mol / L NaOH) a...

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Abstract

The invention discloses a method for fast detecting allergens, belonging to the technical field of detection. The integrated type allergen viable cell detection system integrates cell fixation, allergen irritation and histamine release and determination. The system comprises three cavities consisting of four layers of dimethyl siloxane microchips, wherein the cavities are connected respectively through a silane capillary tube, the cavity at the first layer can be connected with a flow injector or a syringe through a plurality of joints, the cavity at the second layer is an effect cell cavity, and the cavity at the third layer is a histamine reaction basin cavity. After liquid to be detected continuously and fully reacts with effect cells, a solution reaches a histamine reaction basin through a filter membrane and fully reacts with fluorescent derivatization liquid input in advance through reaction liquid injection holes, the generation condition of histamine can be judged under a fluorescent microscope according to fluorescence intensity, and further whether the reaction liquid contains allergens can be judged. The method has excellent sensitivity, fast response and direct visibility, the detection time can be remarkably shortened, the stability is high, and the method has great application potential in detection of allergens in cosmetics, food safety and clinical medicines.

Description

technical field [0001] The invention relates to an analysis system based on living cells and a preparation method thereof, in particular to a living cell analysis technology based on the recognition of amines such as histamine by o-phthalaldehyde. Background technique [0002] Allergy is a common allergic disease that is widely prevalent in the world. Because of its complex symptoms, various types, extensive harm and difficult to cure, it has become a major problem in the medical field for a long time. With the development of the economy and the progress of science and technology, the gradual promotion of genetically modified crops and the increasing popularity of genetically modified foods, food allergens have become one of the hidden dangers of food safety. The significant increase in the incidence of allergies has aroused widespread concern in the society. Allergens refer to antigenic substances that can selectively activate CD4+ Th2 cells and B cells, induce specific Ig...

Claims

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Application Information

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IPC IPC(8): C12M1/00G01N21/64
CPCG01N15/10G01N15/1484G01N21/6486B01L3/502753B01L2400/0406B01L2300/0819B01L2300/0887B01L2200/0668G01N2015/1006G01N15/1023
Inventor 刘永信
Owner TIANJIN MOSBIO SCI & TECH CO LTD
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