OsmolarityLab

NehaKatote
October24th,2015

Introduction:
Theosmolarityofasolutioniseffectivelyitssolutepotential.Asthisaffectstheosmoticpotentialofthe
solutionandthereforetheosmoticpotentialofthecellcontainingthesolution,itwillbeapredictoroftheeffectofthe
environmentonanexposedcell.Ifacellisinahypertonicsolution,thenwaterislikelytomoveoutofthecellby
osmosis,Ifthecellisinahypotonicenvironment,waterwillmoveintothecell.Bythistoken,therewillbenonet
movementofwaterifthecellisisotonicwithitsenvironment.Anunderstandingofthisallowstheestimationofthe
osmolarityofacell,ormorespecifically,thesolutionmakinguptheinternalenvironmentofthecell.

ResearchQuestion:
HowdoestheconcentrationofasaltsolutionaffectthediffusionofwaterinAlliumcepa(red
onion)cells?

Variables:
3
Independentvariable:theconcentrationofthesaltsolution(moldm
)

Dependentvariable:thedistancebetweentheplasmamembraneandthetonoplast

Controlledvariables:
Constantroomtemperature
:thetemperatureaffectsthekineticenergyofparticles,whichinturn,affectsthe
rateatwhichosmosisiscarriedout.
Sametypeofonion
:aredonionisusedbecauseitseasiertoseethedifferentcomponentsoftheonion
cells,asitsstained.However,ifyouuseonewhiteonionandoneredonion,youwontbeabletocompare
thechanges.
Thesurfaceareaofonionepidermisusedineachtrial
:ifonesamplehasalargersurfaceareathanthe
other,thenosmosiswillbecarriedoutatdifferentrates
Volumeofthesaltsolution
:thesamevolumeofsaltsolutionmustbeusedasitcanaffecttheconcentration
gradientsatdifferenttrials.

Materials:
Pieceofredonion,ruler,scalpel,slide,coverslip(&othermaterialsregardingmicroscopicuse),distilled
water,saltwatersolution(atconcentrations:0.5M,1.0M,1.5M,2M),5250mLbeakers,dropper/pipetteand
microscope.

Massofsaltyouneedforeachsolution:
For0.5Mof100mLNaClsolution,youneed2.92gNaCl
For1.0Mof100mLNaClsolution,youneed5.84gNaCl
For1.5Mof100mLNaClsolution,youneed8.77gNaCl
For2.0Mof100mLNaClsolution,youneed11.69gNaCl

Method:
Thefollowingmethodcanbeusedtoobservetheconsequencesofosmosisinredonioncells:
1. Carefullysliceofftheepidermisfromtheofaredonionbulb.Thelayermustbeverythin.
a. Thesamplemustbe5x5mmandmustbeapproximatelythesamesizefromthesameonionat
eachtrial
2. Mountthesampleinadropofdistilledwateronamicroscopeslide,withacoverslip.
3. Observeusingamicroscopeatlowpower.Ifnecessary,observeatmediumpower.Thespaceinsidethe
cellwallwillfillupthecytoplasm,causingtheplasmamembranetochangeshape..
4. Carefullymeasurethedistancebetweentheplasmamembraneandthetonoplast(largevacuolemembrane)
toassessthelevelofwaterlossforaquantifiablegradient.
a. Recordthemeasurementinadatatable
5. Carefullymountanothersampleofepidermisin0.5MNaClsolution.Observesampleatlowpowerand
measurethedistanceasdonepreviously.
6. Repeatthestepsatconcentrations1.0M,1.5Mand2.0M.

EthicsandSafety:
Therearenolivespecimenusedinthisexperimentwhichmeansthattherearenorealethical
issues.However,therearesafetyprecautionsthatyoumusttakeintoconsiderationduringtheexperiment.Asyou
mustsliceaverysmallsampleoftheonionepidermis,itisimportantthatyouusethescalpelverycarefully.Itisalso
importanttokeepthelabareacleanandclearofanywiresfrommicroscopesorotherelectronicdevices.