Biomedical Engineering Reference
In-Depth Information
FIGURE 17.2
Intact.rod.shapes.of.
Salmonella
bacterial.cells.(control).before.treatment.with.MMNs.as.revealed.under.a.TEM.
antibiotics.discs.were.also.applied.to.the.agar.plates,.and.the.plates.were.incubated.at.37°C.
for.24-48.h..The.inhibition.zones.around.the.antibiotic.discs.were.measured.if.observed.
17.2.6 Preparation and Purification of Bacterial Suspension for Examination
and Imaging under Transmission Electron Microscope
Bacterial. cells. were. grown. overnight. in. 10. ml. of. nutrient. broth. in. a. shaker. incubator. at.
37°C,.and.then.1.5.ml.of.the.bacterial.broth.was.added.an.Eppendorf.tube.and.centrifuged.
at.10,000.rpm.for.5.min..The.supernatant.was.discarded,.and.the.pellet.was.washed.twice.
with. sterile. distilled. H
2
O. and. resuspended. using. vortex.. This. bacterial. suspension. was.
viewed.and.examined.as.the.control.under.TEM.(Figure 17.2).
17.2.7 Capturing and/or Binding of Bacterial Cells with Unconjugated (Bare) MMNs
•. The.bacterial.suspension.(200.μl.of.
Salmonella typhimurium
).and.the.MMN.suspen-
sion.(1 ml).were.mixed.in.a.sterile.Eppendorf.tube.
•. Three. replicates. of. the. previous. reaction. mixture. (bacteria-MMN. suspension).
were.used.and.incubated.in.a.shaker.incubator.at.37°C,.where.the.irst.tube.was.
incubated.for.5-10.min,.the.second.tube.for.30.min,.and.the.third.tube.(the.third.
replicate).for.1.h..Then,.15.μl.of.each.tube.was.placed.onto.the.copper.grid.and.then.
dried.in.a.vacuum.desiccator.without.using.any.stain,.and.then.TEM.images.were.
performed.(Figure 17.3)..Figure 17.4.shows.this.bacterial-MMN.aggregate.under.a.
TEM.with.(a).and.without.(b).stain.
•. Two.negative.controls.were.also.used:.200.μl.of.bacterial.suspension.(suspended.in.
deionized.water).and.1.ml.of.MMNs.(suspended.in.deionized.water).
