Two-Phase Microfluidic Systems for High Throughput Quantification of Agglutination Assays, Slides of Engineering

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TWO-PHASE MICROFLUIDIC

SYSTEMS FOR HIGH THROUGHPUT

QUANTIFICATION OF

AGGLUTINATION ASSAYS

ENGR 509 Microfluidics BY: ABDUL BASIT ZIA

OUTLINE 1.1 Immunoassays (Biological background) 1.1.1 Antibody – Antigen reaction 1.2 Diagnostic techniques 1.2.1 Precipitation 1.2.2 ELISA 1.2.3 Fluorescent antibodies 1.2.4 Agglutination 1.3 Droplet microfluidic (Microfluidic Background) 1.3.1 Droplet Generation 1.3.2 Droplet transport 1.3.3 Passive mixing in microfluidics

DIAGNOSTIC TECHNIQUES

Precipitation Reactions Based on two soluble reactants that come together to make one insoluble product, the precipitate Main diagnostic techniques include:

1. Precipitation Reactions
2. ELISA
3. Fluorescent Antibodies
4. Agglutination

DIAGNOSTIC TECHNIQUES- II Enzyme-linked immunosorbent assay (ELISA) The following are key characteristics that make ELISA an ideal method of testing: Versatile Simple Sensitive Quantitative

7 DIAGNOSTIC TECHNIQUES- IV Agglutination Agglutination reactions produce visible aggregates of antibody – antigen complexes when antibodies or antigens are conjugated to a carrier. Antibody molecules are bound to latex beads producing a precipitate of fine particles whole pathogens as a source of antigen uses erythrocytes as the biological carriers

LATEX AGGLUTINATION

DROPLET MICROFLUIDIC – DROPLET GENERATOR

Co-Flowing T-junction Flow focusing

• (^) Dripping: absolute stability
• (^) Jetting : convective stability
• (^) Continuous phase velocity increase leads to dripping to jetting - (^) X<<1 & Ca is high Dripping Viscous shear stress

Interfacial tension

• (^) X=1 & Ca is low Squeezing Single dispersed flow stream Two continuous phase streams No clear cut scaling laws

DROPLET MICROFLUIDIC – DROPLET TRANSPORT Droplets – diameter is smaller than the channel width Plugs – occupies channel’s diameter ∆ 𝑃𝑝𝑙𝑢𝑔𝑠 = 𝑎

𝜇𝑓

𝑊𝐻 )^

𝐿𝑝𝑙𝑢𝑔𝑠 ∙𝑉 (^) 𝑒𝑥𝑡

DEAN FLOW

PASSIVE MIXING

𝑃𝑎𝑟𝑡𝑖𝑐𝑙𝑒 𝑤𝑒𝑖𝑔h𝑡

Questions

OUTLINE OF PRESENTATIONS (1-2) Presentation 1: Introduction 1.1 Immunoassays (Biological background) 1.1.1 Antibody – Antigen reaction 1.2 Diagnostic techniques (ELISA, Agglutination, etc.) 1.3 Role of microfluidics (Microfluidics background) 1.3.1Droplet microfluidic 1.3.2Mixing in microfluidics 1.3.3Agglutination assays in Micro-channels Presentation 2: Agglutination methods and current system 2.1 Mixing methods for agglutinations 2.1.1 Manual 2.1.2 Magnetic 2.1.3 Two-phase serpentine channel 2.1.4 Others 2.2 Basic concept of the desired microfluidic system

Introduction to

concepts

Introduction to

concepts

Literature review

(Lit. Rev.)

Literature review

(Lit. Rev.)

Outline^ Outline

OUTLINE OF PRESENTATIONS (3-4) Presentation 3: Current system for quantification of agglutination assays 3.1 Setup of the current microfluidic system 3.2 Quantification using light scattering 3.2.1 Background 3.2.2 Image processing 3.3 Results of current systems 3.4 Key aspects to focus for achieving high throughput Presentation 4: Achieving high throughput for quantification of agglutination assays. 4.1 Revise key aspects to focus for achieving high throughput 4.2 Results achieved via current improved system

Old Microfluidic

System

Old Microfluidic

System

Results of

Improvements

Results of

Improvements

Lit. Rev.^ Lit. Rev.