116
A. S. Cerda-Kipper and S. Hosseini
capabilities, the presented integrated LOC system could successfully monitor and
control the living cells’ activity (Caputo et al. 2017) (Fig. 5.1).
Dong & Zhao, developed a microfluidic simulator for personalized pathogen
biorecognition by the aim of antimicrobial susceptibility testing (AST) for urinary
tract infection (UTI), a commonly reported bacterial infection (Table 5.1) (Honrado
and Dong 2014). The cell-based LOC system employed the immunosorbent ATPbioluminescence assay (IATP-BLA) as a basic measuring strategy to assess the properties of uropathogenic bacteria. In this study, 13 forms of uropathogenic microbes
were chosen as the target analytes (the IATP-BLA protocol is shown in Fig. 5.2).
The device employed a fiberglass membrane which was sandwiched in between
two polypropylene substrates in order to capture immobilized antibodies on the
membrane. Due to the hydrophilicity of glass fibers, the aqueous media laterally penetrated into the neighboring reaction chambers. The analyte microbes were coupled
with antibodies present on the glass fibers, and were subsequently encapsulated
within a network of calcium alginate gel through the gelation reaction (Dong and
Zhao 2015).
As a disposable biochip (Fig. 5.3), a microfluidic device was fabricated from two
white sheets of polystyrene (PS) processed by laser ablation. A section of Waterman
filter (grade GF-D fiberglass membrane) was firmly clamped between the two
ablated PS layers. Different immunoglobulin Y (IgY) against specific uropathogenic
microbes were immobilized on each zone on the surface of the fiberglass membrane
in chambers. The upper PS component (Air Veins) was used for loading the urine
sample and providing air/oxygen to the cells (Sample Layer). The function of the
lower PS layer, Culture Layer, was to supply culture medium and other reagents to
each reaction chamber. The channel network, Sample Veins, was engraved around
the vertical through holes to lead the urine sample. Through 24 Culture Medium
Veins (CMVs), the culture medium, antibiotic drugs, and ATP-BLA reagents flow
into each reaction chamber. This design was compatible with the standard 384-well
microplate, since the chambers of the microfluid device had the same geometry as
in a traditional 384-well microplate. Compared with conventional microbial culture,
the time of the test cycle was minimized to a few hours or possibly minutes in contrast
to a few days. The microbes were captured by different capture antibodies and were
measured via an ATP bioluminescence assay (ATP-BLA). If provided with sufficient
growth medium, the microbes could survive in the culture compartment for on-chip
AST former to the IATP-BLA measurement. All of the above-mentioned processes
were precisely controlled by a manual valve and pump while the automation of fluidic
manipulation was planned as the future phase of the project. Due to the effectiveness of on-chip IATP-BLA, the system could rapidly identify the commonly known
causative agents of UTIs in the AST within 3–6 h. The authors envisioned that the
medical simulator can be largely used in UTI treatment and could act as a model for
the recognition and treatment of other diseases (Dong and Zhao 2015).
Santangelo et al., designed, fabricated, and tested a 3D-printed chip joined
with silicon photomultipliers (SiPMs) for sensitive and simultaneous detection of
luciferase BL, and for biorecognition of ATP as a model target (Fig. 5.4). The
unibody-LOC (ULOC) 3D printing LOC was previously established for LOC
A. S. Cerda-Kipper and S. Hosseini
capabilities, the presented integrated LOC system could successfully monitor and
control the living cells’ activity (Caputo et al. 2017) (Fig. 5.1).
Dong & Zhao, developed a microfluidic simulator for personalized pathogen
biorecognition by the aim of antimicrobial susceptibility testing (AST) for urinary
tract infection (UTI), a commonly reported bacterial infection (Table 5.1) (Honrado
and Dong 2014). The cell-based LOC system employed the immunosorbent ATPbioluminescence assay (IATP-BLA) as a basic measuring strategy to assess the properties of uropathogenic bacteria. In this study, 13 forms of uropathogenic microbes
were chosen as the target analytes (the IATP-BLA protocol is shown in Fig. 5.2).
The device employed a fiberglass membrane which was sandwiched in between
two polypropylene substrates in order to capture immobilized antibodies on the
membrane. Due to the hydrophilicity of glass fibers, the aqueous media laterally penetrated into the neighboring reaction chambers. The analyte microbes were coupled
with antibodies present on the glass fibers, and were subsequently encapsulated
within a network of calcium alginate gel through the gelation reaction (Dong and
Zhao 2015).
As a disposable biochip (Fig. 5.3), a microfluidic device was fabricated from two
white sheets of polystyrene (PS) processed by laser ablation. A section of Waterman
filter (grade GF-D fiberglass membrane) was firmly clamped between the two
ablated PS layers. Different immunoglobulin Y (IgY) against specific uropathogenic
microbes were immobilized on each zone on the surface of the fiberglass membrane
in chambers. The upper PS component (Air Veins) was used for loading the urine
sample and providing air/oxygen to the cells (Sample Layer). The function of the
lower PS layer, Culture Layer, was to supply culture medium and other reagents to
each reaction chamber. The channel network, Sample Veins, was engraved around
the vertical through holes to lead the urine sample. Through 24 Culture Medium
Veins (CMVs), the culture medium, antibiotic drugs, and ATP-BLA reagents flow
into each reaction chamber. This design was compatible with the standard 384-well
microplate, since the chambers of the microfluid device had the same geometry as
in a traditional 384-well microplate. Compared with conventional microbial culture,
the time of the test cycle was minimized to a few hours or possibly minutes in contrast
to a few days. The microbes were captured by different capture antibodies and were
measured via an ATP bioluminescence assay (ATP-BLA). If provided with sufficient
growth medium, the microbes could survive in the culture compartment for on-chip
AST former to the IATP-BLA measurement. All of the above-mentioned processes
were precisely controlled by a manual valve and pump while the automation of fluidic
manipulation was planned as the future phase of the project. Due to the effectiveness of on-chip IATP-BLA, the system could rapidly identify the commonly known
causative agents of UTIs in the AST within 3–6 h. The authors envisioned that the
medical simulator can be largely used in UTI treatment and could act as a model for
the recognition and treatment of other diseases (Dong and Zhao 2015).
Santangelo et al., designed, fabricated, and tested a 3D-printed chip joined
with silicon photomultipliers (SiPMs) for sensitive and simultaneous detection of
luciferase BL, and for biorecognition of ATP as a model target (Fig. 5.4). The
unibody-LOC (ULOC) 3D printing LOC was previously established for LOC