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S PAT I A L C A L I B R AT I O N O F A N I M AGE USINGIM AGE J

Supposeyouwishtogathermeasurementsfromanimageusingrealvalues(m,miles,etc).Theprocess
involvescalibratingasingleimageagainstknownvalues,thenapplyingthatcalibratedimagetoyourunknown
image.Thisassumes,ofcourse,thatbothimagesarethesamemagnification.Here'sanexample.

Openyouroriginalimageaswellasacalibratedimage.Hereyousee(right)animageofamicroscopestage
micrometer.It'smarkingsare100mand10mapart.

UsetheLineSelectiontooltodrawaselectionlineaknownlength.Herewe'vedrawna400mline:

Withtheselectionlinepresent,select"SetScale"fromtheAnalyzemenu.

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Thisbringsupthefollowingdialogbox:

Justfillinthefields.Ifyoucheckthe"Global"checkbox,thisspatial
calibrationwillbeappliedtoallopenimagewindows.ClickOK.

Theselectyourunknownimage,generateROIs,andAnalyzeParticles.

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InthisexampleImageJcalculatedtheAreaoftheselectedROIsintheoriginalimage.TheResultswindow(R)
showstheareainsquarem(ouroriginalcalibrationscale).

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