Application Of Flow Cytometry Based On Quantum Dot Luminescence Technique In Molecular Detection

Molecular detection is a common detection method,and the detection sample includes DNA,RNA,protein and other small molecular substances.According to the types of the test samples,this kind of detection is collectively referred to as molecular detection.For DNA,RNA,nucleic acid test samples,common techniques include: PCR and DNA sequencing technology,FISH,DNA hybridization imprinting technology,gene chip technology and so on,while in the field of molecular detection in addition to this part also includes FCM,immunohistochemical,ELISA and other detection methods for small molecules.Among them,flow cytometry(FCM)is one of the most advanced cell quantitative analysis techniques and has been widely used in the field of molecular detection due to its advantages of fast,high precision,high accuracy,multiple parameters and high throughput.However,flow cytometry has some application limitations,that is,it can be used to detect cells or biological particles of large molecules,but for small molecules such as proteins and nucleic acids it can't detect directly.This leads to CBA method,that is a multi-label detection technology of trace samples invented by flow giant BD company.It is sold in the form of finished production kits to realize the detection of small molecule samples.However,the cost is very high.How to construct a flow detection model with low cost and high universality to detect small molecular samples?This is the main research content of this manuscript.This study,a general model for flow detection of small molecular samples was introduced,which can be applied to immunohistochemistry,ELISA and other methods besides flow cytometry.The design principle of the detection model was based on enzyme-linked immunity and CBA method in flow cytometry.We all know the CBA method,which is the flow protein quantification technique of trace samples,it uses a series of small and dispersed micros.Heres with fluorescent markers connected specific capture antibodies to capture the objects to be measured in the solution system,and simultaneously determines the quantity of various soluble components in the analysis samples by the different fluorescence intensity of specific micros.The types of samples can be serum,plasma,tears,aqueous humor,saliva,body cavity lavage fluid,cell culture fluid,etc.CBA method has many advantages but is expensive,so the new detection model designed in this study has the advantages of simple operation and high accuracy in addition to the disadvantages of expensive CBA method.Beside,the new model involved in this study is a universal method,which does not require the purchase of a variety of kits,but only requires the purchase of the corresponding two kinds of antibodies to complete the detection.The samples can be soluble cytokines,inflammatory factors,chemokines,enzymes and other protein components,as well as viruses,small molecule nucleic acids and other non-protein components.Only the corresponding antibodies are needed to complete the detection.We first modified sheep anti-rabbit IgG with G5 PAMAM to form a nanoparticle for capturing and supporting corresponding rabbit IgG,and modified sheep anti-mouse IgG with water-soluble cadmium selenide quantum dots to capture mouse IgG and form indicator probe.In this system,the corresponding mouse antibody and rabbit antibody were added to form a sandwich nanocomposite,so that the antigen could be detected in the flow.The detection of the indicators to be measured can be completed only by purchasing the corresponding two antibodies separately.In addition,the sheep anti-rabbit IgG modified by PAMAM and the sheep anti-rat IgG modified by QDs can also be divided into two parts and used separately.For example for immunocytochemistry,western blotting and other methods.In general,this paper detailed introduces a general experimental model can be applied to current detection,and it consists of two parts,respectively is the G5 generation PAMAM modified sheep rabbit IgG part and cadmium selenide quantum dot part modified sheep anti rat IgG,when joining the corresponding rabbit anti rat with resistance can form nano composite sandwich layer type,made by current detection under test indexes.Composition is simple,convenient operation,the model results are accurate,compared with the traditional CBA method can only detect a small number of indicators and the characteristics of the high cost,our model more commonality and two components can be used alone,in addition to the application in flow can also be used in the immunohistochemical,immunocytochemistry method,such as for molecular detection has a high application value.