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入exl=440nll1入e.x2=50Ol
The25thJnternαtionalSymposiumonTransportPhenomena 5-7November2014，KI:abi，Thailand
220C)，for 15rninutesinthecaseofthetubertisueof Jerusalern artichoke or for 10 minutes in the case of the epidermal tis sue of the bulb scale of onion. After that， fluorescenceirnagesofspecirnenat500nrnand440nrnin excitation wavelength were taken respectively for pH.i IrnageJ(7)(8) ofirnage-analysissoftwarewasusedtorequire f1uorescenceintensityratio.First，theobtainedf1uorescence irnagesat500nmand440nrnwereconvertedtothegray scaleones，respectively.Frornthebothgrayscaleirnages， f1uorescence intensity ratio was calculated using the functioninirnageJ.
Next，the area around a nucleus showing strong f1uorescenceintensityinthef1uorescenceirnageat500nrn wasselectedasasubjectofmeasurernent，andthisareawas superposedontheratioirnage.Theaveragevalueoverthis areawasusedasthevalueoff1uorescenceintensityratiofor eachpHi(Figure5).Here，asforone-timeexperirnent，the abovernentionedevaluationofintensityratiowascaITied outfrorntheselectedrnorethanthreareasinthreorfour celsperoneirnage;thatis，oneresultinthisresearchshows theaveragedvalueofmorethan9areasperoneexperirnen.t
Figure6showstherelationshipbetwenpH-valueand f1uorescenceintensityratio，whichwasobtainedinthisway. Here，eachplotshowstheaveragedvalueoftheresultsof thretirnesorrnoreexperirnents.BarisstandardelTor.The relationship betwe en fluorescence intensity ratio and p H showsaproxirnatelylinearone.Forreference，theresults of p H standard solution are also plot ted in Fig.6. The calibrationcurveofpHiagainsttheintensityratioth剖 this researchobtainedwasalsoshowninFig.6.
fsosbesticPoint
Figure4.Relationshipoffluorescenceintensityand excitationwavelength(BCECF-AM).
solution，Alliumcψα L.:1.5μMconcentrationsolution)，then thiscutspecirnenswereirnrnersedfor15rninutes剖 270C.
pHCalibrationCurve ThecalibrationbetwenpHiandfluorescenceintensity
ratiowasperfornedasbelow.Afteraspecirnenwasstained bytheabovef1uorescentreagents，itwasputinpHstandard solutions ofspecific pH (pH 5.5 to 8.5) with 1μM concentration ofNigericinatrornternperature(about
入ex=500nm
• BCECF-.JMμ，IliUtncepaL.) il.BCECF(pHSlandardSolulion)
.BCECF-AMrHelianthusluberos/ls)
λex:ExcitationWavelength
Figure5.Proceduretoobtaintheareaforcalculationof f1uorescenceintensityratio.
Figure6.Fluorescenceintensityratio圃 pHcalibrationcurve (BCECF-AM).
EXPERIMENTALRESULTSANDDISCUSSIONS WithvarγingtherangeofcolingratefromIOC/rninto 10 0oC/rnin，the tuber tis sues of Jerusalern artichoke were coled.pHi-valuewasrneasuredattheintervalof40C・白orn ro orn ternperature (about 220 C) up to the ternpera加 re when
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