Page 370 - konishipaper

24 工Ninagawaetロ1./Cryobiology73(201旬 20-29
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2.5.D祈 nitionofthedegre eofcel ldゆ rmation
Thedegreofceldeformation(e)indicateshowmuchacelhas beendeformedasaresultofintracelularicecrystals，andwas measuredbycalculatingtheareaofcelsbefore(Ab)andimmedi- atelyafter(Ac)intracelularfrezing.Theewasthencalculatedas:
Ac-Ab E二二一一ーで一一一
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2.6. Evaluationofthegrowingrateofintracel lularicecrystals
The growing rate of intracel lular ice crystals was evaluated
basedontheareavelocityofintracelularicecrystals，bymeasuring theareaofanintracelularicecrystalevery2msfromthetimeofits apearanceuntil10mshadelapsed.
2.7.Statisticalanalyses
Thedegreofsupercolingwaspresentedtheaveragevalue obtainedfrom10specimensateachcolingrate.IFbehaviorswere analyzedusingfourmovingimagesobtainedduringintracelular frezingateachcolingrate.Theseimageswereusedtoexamine thelocationsatwhichintracelularicecrystalsapearedandthe formationpaternsofintracelularicecrysta.lSinceintracelular frezinginthesameplanttisueocuredmorefrequentlywithan increaseincolingratedespitethesamecapturetime(2s)being used，thesamplesizealsoincreasedwithanincreaseincoling rate.
Toevaluatethedegre eofcel ldeformation(e)，thegrowingrate， andthegrainsizesattheedgesofintracelularicecrystals，atleast 10 cel ls were randomly sampled acros s the four moving images in
ElapseIctime企omthestartofintracelularfrezing.
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(C)U.7ゅ8吟8ゅ9吟 (D) 回 Formationpa託ern(A) 闇 Fonationpatern(B) ・Formationpatern(C)
10吟o.()ゅC吟。ゆ (B1Uゅ6ゅ e吟@ 司ゆ}令
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50
畠 40 ω
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色 10 O
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-10 -100 Coolingrate[OC/min]
Fig.5.SchematiciIlustrationoftheformationpaternsofintracelularicecrystalsintheepidermaltisuesofstrawberygeranium(S似 俳 句 日 ωslonifera仁urtis)leaves.(A)Schematic Formationpa仕ernA，whereintracelularicecrystalsformedaroundtheoutlineofthecel.(B)FormationpaternB，whereintracelularicecrystalsformedacrosthecenterofthe cel.l(C)FormationpaternC，whereintracelularicecrystalsexpandedfromthecenterofthecel.l(0)Proportionofcelsexhibitingeachtypeoformationpaternatcolingratesof -1oCjmin(n=15)，-10oCjmin(n=28)，and-10oC/min(n=45).(A-C)Smalarows:directionofprogresintheformationofintracelularicecrystals.
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