Biomedical Engineering Reference
In-Depth Information
#2
#3
#1
FOV
=
120
µ
m
FOV
=
125
µ
m
FOV
=
20
µ
m
#5
#4
#6
FOV
=
50
µ
m
FOV
=
50
µ
m
FOV
=
50
µ
m
FIGuRE 5.6
Visualization.of.the.specimen-induced.distortions.at.an.NA.of.0.6:.simulation.of.imaging.a.regular.
grid.placed.underneath.the.specimen..he.measured.distortions.are.plotted.as.a.deviation.from.the.regular.mesh.
positions.of.the.grid.and.are.enlarged.by.a.factor.of.ive..he.specimen.numbers.refer.to.Table.4.2.of.Chapter.4,.and.
the.ield.of.view.(FOV).covered.by.the.scan.is.also.given.for.each.specimen.
5.6 Summary and conclusion
A. method. to. obtain. quantitative. data. for. the. spatial. measurement. inaccuracies. due. to. the. specimen-
induced.aberrations.has.been.presented..he.approach.uses.interferometry.to.record.wavefront.aberra-
tion.data.from.the.pupil.plane.of.the.microscope.lens.to.extract.information.about.local.distortion..To.
the.best.of.our.knowledge,.this.is.the.irst.attempt.to.quantify.the.efects.of.specimen-induced.image.
distortion.in.microscopy..his.information.is.usually.not.accessible.and.cannot.be.observed.directly.in.
a.standard.epi-type.microscope..However,.our.results.show.that.the.specimen-induced.distortions.can.
signiicantly.inluence.the.spatial.measurement.accuracy.that.can.be.achieved.with.an.otherwise.perfect.
and.calibrated.microscope..Investigators.should.consider.these.efects.whenever.spatial.measurements.
of.high.accuracy.are.required.
Acknowledgment
It.is.noted.that.these.results.have.also.been.presented.in.the.publication.(Schwertner.et.al.,.2007).
references
Pawley,.J..(2002)..Limitations.on.optical.sectioning.in.live-cell.confocal.microscopy..
Scanning 24
,.241-246.
Schwertner,. M.,. M.. J.. Booth,. and. T.. Wilson. (2007).. Specimen-induced. distortions. in. light. microscopy..
J. Microsc
..228(1),.97-102.
Wilson,.T..and.C..Sheppard.(1984)..
heory and Practice of Scanning Optical Microscopy
..Academic.Press,.
London.
