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Kiichi Sato,† Manabu Tokeshi,† Tamao Odake,† Hiroko Kimura,‡ Takeshi Ooi,§ Masayuki Nakao,§ and PowerPoint Presentation
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Kiichi Sato,† Manabu Tokeshi,† Tamao Odake,† Hiroko Kimura,‡ Takeshi Ooi,§ Masayuki Nakao,§ and - PowerPoint PPT Presentation


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Integration of an Immunosorbent Assay System : Analysis of Secretory Human Immunoglobulin A on Polystyrene Beads in a Microchip. Kiichi Sato,† Manabu Tokeshi,† Tamao Odake,† Hiroko Kimura,‡ Takeshi Ooi,§ Masayuki Nakao,§ and Takehiko Kitamori*,†, |. Introduction. Integrated immunosorbent assay

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Integration of an Immunosorbent Assay System:Analysis of Secretory Human Immunoglobulin A onPolystyrene Beads in a Microchip

Kiichi Sato,† Manabu Tokeshi,† Tamao Odake,† Hiroko Kimura,‡ Takeshi Ooi,§ Masayuki Nakao,§ and

Takehiko Kitamori*,†,|

introduction
Introduction
  • Integrated immunosorbent assay
  • Microchip fabrication
  • Experiment

antigen and antibody –colloidal gold conjugated

apparatus

  • Result
  • Discussion
integrated immunosorbent assay
Integrated immunosorbent assay

*Conventional immunosorbent assay

- need long time

- many expensive reagent

- troublesome procedure

Integrated immunosorbent assay

reduce assay time < 24 h

simple operation

experiment
Experiment
  • Antigen and antibody-colloidal gold conjugate

reaction : 50ul antiserum + 5ml of pH adjusted gold suspension

(at room temp. for 30 min)

blocking : 25ul 30% BSA in PB (at 4 Co overnight incubation)

maximum absorbance at 525nm

1 week preserve without loss of activity

  • Adsorption of s-IgA on polystyrene bead

50ul suspended bead + 0.5ml Ag solution

(at room temp. for 1h and at 4 Co overnight incubation)

blocking with casein solution

(at room temp. for 1h )

washing PB

tlm thermal lens microscope
TLM (Thermal Lens Microscope)

진공상태나Ar가스 분위기 상태에서

재료를 고온으로 가열하여 thermaletching및 gas etching에 의하여

나타나는 미세조직의 변화를 관찰하고 그 온도에서 시료에 일어나는 여러 가지 변화를 관찰.

금속, 세라믹 고분자 및 복합재료에서의 (1) 가열·냉각하는 동안의 상변태,

(2)고온에서의 결정립의 성장,

(3) 표면확산현상, (4) 고온 소성변형과 파단, (5) 고온취성, (6) 금속 및 비금속의 소결, (7)부식과정 등의 연구에 이용

excitation beam :

Ar + laser 514.5nm

probe beam : He-Ne laser 632nm

good point of this methods
Good point of this methods

Specific interface ratio ( S / V )

50ul micro plate ( 0.65mm diameter )

==== 13cm-1

Microchannel

(11bead 100um * 100um * 200um )

==== 480cm -1

37 time larger than micro plate!!

distance to reaction

Microplate : 1.5mm

Microchannel : 20um

5600 times shorter !!

consequently reduce time to 10 min

antigen adsorption
Antigen adsorption

-- Injection polystyrene bead

-- s IgA sol (10ug/ml) injected

-- incubation

-- blocking casein

-- AbCG sol injected for 10min

almost all of reaction was

finished in 20 min

immunoassay in microchip
Immunoassay in microchip

Adsoption sample : 20 min

blocking : 10 min

Ag-Ab reaction : 10 min

washing : 1min

total less than 1 hour !!

discussion
Discussion
  • Simple operation
  • Remove troublesome procedure
  • Reduce expensive reagent
  • Association with automated system
  • Application with DNA Computing