Biomedical Engineering Reference
In-Depth Information
AO correction at 145 µ m
µ m
µ m
0.5
0.4
0.3
0.2
0.1
0
2.5
2
1.5
1
- 0.1
- 0.2
- 0.3
- 0.4
0.5
0
5
10 15 20
Zernike terms
25 30
35 40
2
4
6
8
10
12
DM shape
FIGuRE 12.15 he.Zernike.terms.and.resulting.DM.shape.for.the.optical.aberration.correction.for.the.mouse.
bone.cavity.imaging.at.145.μm.below.the.surface.
AO correction at 184 µ m
µ m
0.6
0.4
µ m
3.5
3
0.2
0
2.5
2
- 0.2
1.5
- 0.4
1
- 0.6
0.5
- 0.8
0
5
10 15 20
Zenike terms
25 30 35 40
2
4
6
8
10
12
DM shape
FIGuRE 12.16 he.Zernike.terms.and.resulting.DM.shape.for.the.optical.aberration.correction.for.the.mouse.
bone.cavity.imaging.at.184.μm.below.the.surface.
Figures.12.15.and.12.16.show.the.resulting.DM.shapes.and.Zernike.terms.for.the.aberration.correc-
tion.at.the.two.diferent.depths..he.correction.shape.is.largely.afected.strongly.by.the.bone.structure.
and. depth.. As. predicted,. the. considerable. spherical. aberration. is. corrected. in. the. beam. path. at. both.
locations..At.184.μm,.the.irst.spherical.aberration.amplitude.is.0.41.μm;.however,.Zernike.term.14.is.
the.most.signiicant.aberration.at.−0.6.μm.RMS..At.145.μm,.astigmatism.stands.out.at.0.48.μm.RMS.
12.6 critical Discussion
he.imaging.depth.is.limited.at.about.200.μm.for.the.GFP.mouse,.which.is.about.half.the.desired.imag-
ing.depth.for.the.entire.mouse.skull.bone..Deeper.detection.is.not.achievable.because.of.the..diminishing.
luorescence. signal.. Higher. illumination. intensity. can. be. used. to. increase. the. luorescence. intensity.
signal..However,.such.an.option.is.limited,.to.avoid.thermal.damage.of.the.sample.that.is.being.imaged.
he. collected. luorescence. signal. can. also. be. increased. by. enlarging. the. numerical. aperture. of. the.
collection.optics..Combs.et.al..use.parabolic.mirrors,.combined.with.the.imaging.objective,.to.collect.
the.solid.angle.around.the.excitation.spot.when.imaging.the.sample.[47]..he.enlarged.collection.angle.
allows.twofold.improvement.of.the.luorescence.signal.in.their.prototype.instrument.
In. an. attempt. to. demonstrate. AO. in. deeper. bone-marrow. imaging,. we. used. an. unconventional.
approach.to.create.a.strong.luorescence.signal.in.a.deep.layer.(200.μm.or.deeper.below.the.surface).in.
the.mouse.skull.bone.structure.for.AO.correction.
 
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