IIF CHARACTERISTICS OF OYSTER EMBRYOS AND EGGS DETERMINED BY A FEEDBACK CONTROLLED DIRECTIONAL CRYOMICROSCOPE Ta-Te Lin, Ken Lung Department of Agricultural.
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IIF CHARACTERISTICS OF OYSTER EMBRYOS AND IIF CHARACTERISTICS OF OYSTER EMBRYOS AND
EGGS DETERMINED BY A FEEDBACK EGGS DETERMINED BY A FEEDBACK
CONTROLLED DIRECTIONAL CRYOMICROSCOPECONTROLLED DIRECTIONAL CRYOMICROSCOPE
Ta-Te Lin, Ken Lung
Department of Agricultural Machinery Engineering,
National Taiwan University,
Taipei, Taiwan, R.O.C.
OBJECTIVES
MATERIALS AND METHODS
RESULT AND ANALYSIS
CONCLUSIONS
OBJECTIVESOBJECTIVES
To design and construct a feedback controlled directional cryomicroscope system.
To determine the IIF characteristics of oyster embryos and eggs at subfreezing temperatures.
To compare and analyze the IIF characteristics of oyster embryos and eggs with a stochastic model.
MATERIALS AND METHODSMATERIALS AND METHODS
THE DIRECTIONAL CRYOMICROSCOPE SYSTEM PREPARATION OF OYSTER EGGS AND EMBRYOS EXPERIMENTS
TESTS ON THE CRYOMICROSCOPE SYSTEM PERFORMANCE
OBSERVATION OF IIF PROCESS DURING FREEZING
MATERIALS AND METHODSMATERIALS AND METHODS
THE DIRECTIONAL CRYOMICROSCOPE SYSTEM PREPARATION OF OYSTER EGGS AND EMBRYOS EXPERIMENTS
TESTS ON THE CRYOMICROSCOPE SYSTEM PERFORMANCE
OBSERVATION OF IIF PROCESS DURING FREEZING
MATERIALS AND METHODSMATERIALS AND METHODS
THE DIRECTIONAL CRYOMICROSCOPE SYSTEM PREPARATION OF OYSTER EGGS AND EMBRYOS EXPERIMENTS
TESTS ON THE CRYOMICROSCOPE SYSTEM PERFORMANCE
OBSERVATION OF IIF PROCESS DURING FREEZING
THE DIRECTIONAL CRYOMICROSCOPE SYSTEM PREPARATION OF OYSTER EGGS AND EMBRYOS EXPERIMENTS
TEST ON CRYOMICROSCOPE PERFORMANCE OBSERVATION OF IIF PROCESS DURING FREEZING
MATERIALS AND METHODSMATERIALS AND METHODS
RESULTSRESULTS
TIME AND TEMPERATURE DEPENDENCE OF IIF THE STOCHASTIC MODEL FOR IIF PHENOMENON SIMULATION AND ANALYSES
OC BELOW FREEZING
12 14 16 18 20 22 24 26 28 30
PR
OB
AB
ILIT
Y O
F II
F
0
10
20
30
40
50
60
70
80
90
100
EGGS
EMBRYOS
Comparison on the probability of IIF for oyster eggs and embryos at subfreezing temperatures
oC BELOW FREEZING
16 18 20 22 24 26 28 30
PR
OB
AB
ILIT
Y O
F II
F (
%)
0
10
20
30
40
50
60
70
80
90
100
TOTAL IIF
INSTANTANEOUS IIF
TIME-DEPENDENT IIF
Simulation on the relative probability of instantaneous IIF and time-dependent IIF mechanisms for oyster embryos.
oC BELOW FREEZING
18 20 22 24 26 28 30
PR
OB
AB
ILIT
Y O
F II
F (
%)
0
10
20
30
40
50
60
70
80
90
100
TOTAL IIF
INSTANTANEOUS IIF
TIME-DEPENDENT IIF
Simulation on the relative probability of instantaneous IIF and time-dependent IIF mechanisms for oyster eggs.
TIME (MIN)
0 1 2 3 4
TE
MP
ER
AT
UR
E (
o C)
-40
-35
-30
-25
-20
-15
-10
-5
0
5
10
-30oC
-20oC
-10oC
Test on the isothermal temperature control of the directional cryomicroscope.
TIME (MIN)
0 1 2 3 4 5 6
TE
MP
ER
AT
UR
E (
o C)
-40
-35
-30
-25
-20
-15
-10
-5
0
5
10
-40oC/min
-20oC/min
-10oC/min
Set point temperature and the experimental temperature measured with the thermocouple under linear cooling rate controlled protocol.
Comparison on the IIF experimental results of oyster embryos and the fitted curves of the modified stochastic model
TIME (min)0 1 2 3 4 5 6 7 8
PR
OB
AB
ILIT
Y O
F II
F (
%)
0
10
20
30
40
50
60
70
80
90
100
-20oC
-24oC
-25oC-30oC
-23oC
-22oC
-15oC
Comparison on the IIF experimental results of oyster eggs and the fitted curves of the modified stochastic model
TIME (min)0 1 2 3 4 5 6 7 8
PR
OB
AB
ILIT
Y O
F II
F (
%)
0
10
20
30
40
50
60
70
80
90
100
-20oC
-24oC
-22oC
-25oC-30oC
TIME (min)0 1 2 3 4 5 6 7 8
PR
OB
AB
ILIT
Y O
F II
F (
%)
0
10
20
30
40
50
60
70
80
90
100
-20oC
-24oC
-25oC-30oC
-23oC
-22oC
-15oC
Comparison on the IIF experimental results of oyster embryos and the fitted curves of the stochastic model
TIME (min)0 1 2 3 4 5 6 7 8
PR
OB
AB
ILIT
Y O
F II
F (
%)
0
10
20
30
40
50
60
70
80
90
100
-20oC
-24oC
-22oC
-25oC-30oC
Comparison on the IIF experimental results of oyster eggs and the fitted curves of the stochastic model
TIME (MIN)0 1 2 3 4 5 6 7 8
PR
OB
AB
ILIT
Y O
F II
F (
%)
0
10
20
30
40
50
60
70
80
90
100
-20oC
-24oC
-22oC
-25oC-30oC
Time and temperature dependence of IIF process for oyster eggs.
TIME (MIN)0 1 2 3 4 5 6 7 8
PR
OB
AB
ILIT
Y O
F II
F (
%)
0
10
20
30
40
50
60
70
80
90
100
-20oC
-24oC
-25oC-30oC
-23oC
-22oC
-15oC
Time and temperature dependence of IIF process for oyster embryos.
THE STOCHASTIC MODEL
P{ IIF } = { 1 - exp[ - ( T / T0 )] }
+ { 1 - [1 - exp( -(T / T0))]}‧[1 - exp(-t / t0)]
= 1 - exp [ - ( T / T0 )- (t / t0)]
1 / t0 = (T) = ( T / t0 )
(Pitt et al., 1991) INSTANTANEOUS IIFTIME-DEPENDENT IIF
(Modified model)
P{ IIF } = { 1 - exp[ - ( T / T0 )] }
+ { exp[ - ( T / Ta )] - exp[ - ( T / T0 )
] }
‧[ 1 - exp( -t / t0 ) ]
1 / t0 = (T) = ( T / t0 )
THE STOCHASTIC MODEL
THE DETERMINED KINETIC PARAMETERS
PARAMETERS OYSTEREMBRYOS
OYSTEREGGS
DROSOPHILA
EMBRYOS*
T0 24.4C 37.2C 20.4 4.20 3.20 2.94
0 25.6 25.0 65.5 3.24 8.67 2.59
*IN BD.20 SOLUTION (Pitt et al., 1991)
THE DETERMINED KINETIC PARAMETERS
(Modified model)
PARAMETERS OYSTEREMBRYOS
OYSTEREGGS
T0 24.4C 37.2C 4.20 3.20
0 16.0 25.3 1.04 4.93Ta 19.9C 20.8C 7.10 15.8
THANK YOU
謝 謝
Driven bystepping motor
Fixed horizontally Samples
SCHEMATIC DIAGRAM OF THE DIRECTIONAL CRYOMICROSCOPE
CONCLUSIONSCONCLUSIONS
An integrated, feedback controlled directional cryomicroscope system was developed.
IIF temperatures of oyster eggs and embryos were determined using the cryomicroscope system. The IIF process of oyster embryos and eggs was found to be both time and temperature dependent.
The median IIF temperature of embryos and eggs were determined to be -22.5C and -21.0C, respectively. The transition temperature zone of IIF appeared to be broader for embryos than that of eggs.
The IIF data were further analyzed with a stochastic model and the kinetic parameters were determined.
LN2
STEPPINGMOTOR
HEAT SINK HEAT SOURCE
GLASS SLIDE
COLD STAGE BASE
LIMITSWITCH
COLD STAGE
SIGNALPROCESSOR
DRIVER
WORM GEAR
TEMPERATURESIGNAL
TEMPERATURESIGNAL
HEAT SINKCONTROLLER
HEAT SOURCECONTROLLER
RS232
HEATING CURRENT
HEATING CURRENT
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