3. Software for image acquisition: Nucline (Mediso, Hungary).
4. Software for 3D image reconstruction: IVS image processing
(Invicro, USA).
5. Software for image analysis: PMOD 3.7 (PMOD Technologies
LLC, Switzerland).
2.2 Cooling
Equipment for CT
Measurement of
Cryopreserved
Samples
1. A liquid nitrogen dewar (about 2 L of volume) containing a
copper coil of about 10 mm inner diameter and the maximum
length and width possible to fit inside the dewar (see Note 2).
2. A compressed nitrogen gas tank with a pressure-reducing valve
to control the nitrogen gas flow.
3. A polypropylene tube of 20 mm inner diameter from the
compressed nitrogen gas outlet to the copper coil described
above (room temperature gas) and a polystyrene tube of
20 mm inner diameter and 20 mm of thickness from the copper
coil to the insulating container described below (cooled nitrogen gas), with dimensions according to the carbon fiber CT
bed (see Note 3).
4. A cryochamber for holding the sample to image, made of a
cylindrical insulating container of polystyrene (60 mm outer
diameter maximum), with several tubes of 1.5 mm inner diameter connected at the end of the tube system described above,
through which the cooled nitrogen gas passes (see Figs. 1
and 2).
5. A type K thermocouple, located inside the insulating container
described above and connected to a data logger (TC-08 Temperature Data Logger, Picolog).
6. A small computer (e.g., notebook) to install the Picolog software to monitor the temperature during the CT
measurements.
2.3 Solutions and
Containers
1. Isotonic phosphate-buffered solution (PBS) as solvent,
prepared from deionized water (NaCl 8 g/L, KCl 0.2 g/L,
Na 2 HPO 4 1.44 g/L, KH 2 PO 4 0.24 g/L) and adjusted to
pH 7.4.
2. Dimethyl sulfoxide as cryoprotective agent in concentrations
ranging from 5% to 70% (v/v).
3. There are two conditions that are going to influence the selection of the material and the dimensions of the sample containers. The material used for containers has to be transparent to Xrays, in order to avoid interferences in the images (see Note 4).
The dimensions of the containers are limited by the size of the
CT bed, and also depend on the size of the samples to be
analyzed (see Note 5).
X-Ray Computed Tomography
319
4. Software for 3D image reconstruction: IVS image processing
(Invicro, USA).
5. Software for image analysis: PMOD 3.7 (PMOD Technologies
LLC, Switzerland).
2.2 Cooling
Equipment for CT
Measurement of
Cryopreserved
Samples
1. A liquid nitrogen dewar (about 2 L of volume) containing a
copper coil of about 10 mm inner diameter and the maximum
length and width possible to fit inside the dewar (see Note 2).
2. A compressed nitrogen gas tank with a pressure-reducing valve
to control the nitrogen gas flow.
3. A polypropylene tube of 20 mm inner diameter from the
compressed nitrogen gas outlet to the copper coil described
above (room temperature gas) and a polystyrene tube of
20 mm inner diameter and 20 mm of thickness from the copper
coil to the insulating container described below (cooled nitrogen gas), with dimensions according to the carbon fiber CT
bed (see Note 3).
4. A cryochamber for holding the sample to image, made of a
cylindrical insulating container of polystyrene (60 mm outer
diameter maximum), with several tubes of 1.5 mm inner diameter connected at the end of the tube system described above,
through which the cooled nitrogen gas passes (see Figs. 1
and 2).
5. A type K thermocouple, located inside the insulating container
described above and connected to a data logger (TC-08 Temperature Data Logger, Picolog).
6. A small computer (e.g., notebook) to install the Picolog software to monitor the temperature during the CT
measurements.
2.3 Solutions and
Containers
1. Isotonic phosphate-buffered solution (PBS) as solvent,
prepared from deionized water (NaCl 8 g/L, KCl 0.2 g/L,
Na 2 HPO 4 1.44 g/L, KH 2 PO 4 0.24 g/L) and adjusted to
pH 7.4.
2. Dimethyl sulfoxide as cryoprotective agent in concentrations
ranging from 5% to 70% (v/v).
3. There are two conditions that are going to influence the selection of the material and the dimensions of the sample containers. The material used for containers has to be transparent to Xrays, in order to avoid interferences in the images (see Note 4).
The dimensions of the containers are limited by the size of the
CT bed, and also depend on the size of the samples to be
analyzed (see Note 5).
X-Ray Computed Tomography
319