Biomedical Engineering Reference
In-Depth Information
Blue
laser
Green
SC
Red
laser
Crimson
SC
Sample
Ob
IX71
F1
L1
L4
D
L2
SHWS
F2
F3
BS
L3
DM
FIGuRE 17.5
AO.wide-ield.microscope.setup..DM,.SHWS,.488.nm.laser.(blue.laser),.green.lourescent.science.
camera.(green.SC),.HeNe.632.8.nm.laser.(red.laser),.crimson.lourescent.science.camera.(Crimson.SC)..L1,.L2,.L3,.
and.L4.are.180,.85,.275,.and.225.mm.focal.length.lenses,.respectively..Fold.mirror.D.helps.to.bring.the.optical.path.
into.alignment.for.the.SHWS.
Green
SC
Crimson
SC
SHWS
DM
FIGuRE 17.6
Adaptive.optics.wide-ield.microscope.set.up.with.Olympus.IX71.inverted.microscope.and.Boston.
Micromachines.DM.
laser).through.a.confocal.illuminator.(not.shown).onto.the.optical.path.for.excitation.of.the.crimson.refer-
ence.beads.(Azucena.et al..2010,.2011a)..his.confocal.illuminator.allows.us.to.illuminate.a.single.crimson.
reference.bead.to.create.a.single.difraction-limited.spot..he.beam.splitter.(BS).lets.90%.of.the.emitted.
light.coming.from.the.crimson.reference.beads.go.to.the.SHWF.sensor.for.wavefront.measurement.and.
10%.for.imaging.in.the.Crimson.Science.Camera.(Crimson.SC)..he.SHWF.sensor.is.composed.of.a.44.
×.44..element.lenslet.array.each.with.a.focal.length.of.24.mm.and.a.diameter.
d
LA
.=.328.μm.(AOA.Inc.,.
Cambridge,.MA),.and.a.cooled.CCD.camera.(Roper.Scientiic,.Acton,.NJ)..Figure.17.6.shows.the.setup.on.
the.optical.table,.highlighting.a.few.of.the.components.like.the.DM,.science.cameras,.and.SHWF.sensor.
17.2.1 Fluorescent Microsphere reference Beacons (Guide Stars)
We. have. developed. a. microsphere. injection. process,. which. works. very. well. for. introducing. reference.
beacons,. or. guide. stars,. into. live. embryos.. his. process. does. not. harm. the. embryos. as. they. have. the.
ability. to. heal. the. wound. around. the. injection. site.. Using. a. 1:1000. concentration. microsphere. solu-
tion.assures.that.there.will.be.a.microsphere.within.10.μm.of.the.center.of.the.injection.discharge..he.
microspheres.spread.in.a.random.manner.around.the.discharge.site.and.can.usually.be.found.to.spread.
