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Microfluidics for Microarrays

Run Assays on a Microarray fully-automatically

Video of onsite.flow cartridge with heating

Our single-use microfluidic cartridges are equipped with integrated functionalities such as
  • pumping,
  • heating and
  • reagent storage.
The functionalities of the cartridges can be controlled electronically just by software. No external pumps and no tubing are required.

Program an assay, pipette sample and reagents into the reservoirs and press the „Start“ button.

 

Microarray Integration

Polymer foil with spotted microarray is placed on sensor channel of na.flow cartridge.

Both, microarrays spotted on glass and polymer slides can be used with our cartridges. We can provide functionalized polymer foils with low autofluorescence which come pre-cut to fit already to a certain cartridge.

Typically, the microarrays need to be designed to fit into a flow channel  (=sensor area) of 2 to 3 mm width, which results in arrays of up to around 100 spots. With smaller spot size and pitch, also larger arrays are possible.

Example 1: Hybridization on a DNA Microarray

Species Identification and Antibiotic Resistance Detection

microarray_hybridization.jpgFalse-color fluorescence image of a microarray after hybridization of amplified and labeled bacterial DNA
 
 
Samples were prepared by lysis of clinical isolates and amplification of 16S rDNA (including labelling), followed by a purification and a digestion step to randomly shorten the DNA to smaller fragments suitable for hybridization. After filling reagents and DNA sample into the reservoirs of a cartridge, hybridization assays as shown in the following table were run in the cartridges fully-automatically with a total time of 24-39 min.

 #DescriptionTime  [min]
 1Heating sensor area / hybridization chamber with microarray to 48°C1
 2Pumping pre-hybridization buffer1
 3Pumping reservoirs with DNA sample and hybridization buffer simultaneously towards sensor area15-30
 4Switching off heaters to cool-down sensor area1
5Pumping three reservoirs with wash buffers2 + 2 + 2

Example 2: Detection of CRP

Sandwich Immunoassay with immobilized antiCRP and different Control Antibodies

False-color fluorescence image and layout of a microarray after sandwich immunoassay on a flex.flow cartridge. Concentration: 1ug/ml CRP

To detect the bound CRP a monoclonal antiCRP(mouse) is used followed by an antiMouse, Cy-5 labeled antibody as marker.
For the experimental set-up, on a flex.flow cartridge the pumping sequence  and the allocation of the different reagents, washing buffers and the sample was chosen as follows:

Reagents in the reservoirs
Reservoir 1: CRP sample
Reservoir 2: antiMouse (Cy5-labeled, sheep)
Reservoir 3: milk powder (3%, PBS)
Reservoir 4: deionized water
Reservoir 5: milk powder (3%, PBS)
Reservoir 6: antiCRP (monoclonal, mouse)

Pumping Sequence:
Step 1: Reservoir 3, 10 s
Step 2: Reservoir 1, 30 s
Step 3: Reservoir 5, 30 s
Step 4: Reservoir 6, 30 s
Step 5: Reservoir 3, 20 s
Step 6: Reservoir 2, 20 s
Step 7: Reservoir 4, 30 s