the insulating containers (see Figs. 1 and 2), and wait for about
10 min to cool it down, depending on the size of the sample
(see Note 13).
7. Check that the nitrogen gas pressure is the required one and
the temperature is down to À140
C, and refill the liquid
nitrogen of the dewar.
8. Repeat the steps 4–7 of Subheading 3.1. Before the CT acquisition of each sample, wait until the temperature is cooled
down to À140
C, wait for 10 min to cool it down, check the
pressure of the nitrogen gas, and refill the liquid nitrogen of the
dewar.
9. Once all the samples have been imaged, close the valve of
nitrogen gas, and then turn off the Picolog software.
10. Repeat the steps 8–12 of Subheading 3.1, and obtain the
calibration curve for the samples imaged below À140
C (see
Fig. 5 as an example).
3.3 CT Acquisition of
the Biological Samples
After Cryopreservation
1. Proceed with your routine or any other cryopreservation protocol of the samples you wish to analyze under the CT. Store
the cryopreserved samples in liquid nitrogen or any other usual
storage.
2. Place the cooling system on the CT device, as can be seen in
Fig. 1. Check that the insulating container and the holder you
use are suitable for the container of your samples (see Fig. 2).
3. Repeat the steps 2–5 of Subheading 3.2.
4. When the temperature has cooled down to À140
C, place the
first sample to image inside the insulating container. Make sure
you do this operation very quickly in order to avoid any devitrification of the sample.
5. Check that the nitrogen gas pressure is the required one and
the temperature is down to À140
C, and refill the liquid
nitrogen of the dewar.
6. Repeat the steps 4–7 of Subheading 3.1. Before the CT acquisition of each sample, wait until the temperature is cooled
down to À140
C, check the pressure of the nitrogen gas,
and refill the liquid nitrogen of the dewar.
7. Once all the samples have been imaged, close the valve of
nitrogen gas, and turn off the Picolog software.
3.4 CT Acquisition of
the Biological Samples
After Rewarming
1. Place the cooling system on the CT device, as can be seen in
Fig. 1.
2. Proceed with your routine or any other warming protocol of
the cryopreserved samples you wish to analyze under the
X-Ray Computed Tomography
325
10 min to cool it down, depending on the size of the sample
(see Note 13).
7. Check that the nitrogen gas pressure is the required one and
the temperature is down to À140
C, and refill the liquid
nitrogen of the dewar.
8. Repeat the steps 4–7 of Subheading 3.1. Before the CT acquisition of each sample, wait until the temperature is cooled
down to À140
C, wait for 10 min to cool it down, check the
pressure of the nitrogen gas, and refill the liquid nitrogen of the
dewar.
9. Once all the samples have been imaged, close the valve of
nitrogen gas, and then turn off the Picolog software.
10. Repeat the steps 8–12 of Subheading 3.1, and obtain the
calibration curve for the samples imaged below À140
C (see
Fig. 5 as an example).
3.3 CT Acquisition of
the Biological Samples
After Cryopreservation
1. Proceed with your routine or any other cryopreservation protocol of the samples you wish to analyze under the CT. Store
the cryopreserved samples in liquid nitrogen or any other usual
storage.
2. Place the cooling system on the CT device, as can be seen in
Fig. 1. Check that the insulating container and the holder you
use are suitable for the container of your samples (see Fig. 2).
3. Repeat the steps 2–5 of Subheading 3.2.
4. When the temperature has cooled down to À140
C, place the
first sample to image inside the insulating container. Make sure
you do this operation very quickly in order to avoid any devitrification of the sample.
5. Check that the nitrogen gas pressure is the required one and
the temperature is down to À140
C, and refill the liquid
nitrogen of the dewar.
6. Repeat the steps 4–7 of Subheading 3.1. Before the CT acquisition of each sample, wait until the temperature is cooled
down to À140
C, check the pressure of the nitrogen gas,
and refill the liquid nitrogen of the dewar.
7. Once all the samples have been imaged, close the valve of
nitrogen gas, and turn off the Picolog software.
3.4 CT Acquisition of
the Biological Samples
After Rewarming
1. Place the cooling system on the CT device, as can be seen in
Fig. 1.
2. Proceed with your routine or any other warming protocol of
the cryopreserved samples you wish to analyze under the
X-Ray Computed Tomography
325