Improvements in the technique of vascular perfusion-fixation employing a fluorocarbon-containing perfusate and a peristaltic pump controlled by pressure feedback
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Improvements in the technique of vascular perfusion-fixation employing a fluorocarbon-containing perfusate and a peristaltic pump controlled by pressure feedback. / Rostgaard, J; Qvortrup, Klaus; Poulsen, Steen Seier.
In: Journal of Microscopy, Vol. 172, No. Pt 2, 11.1993, p. 137-51.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Improvements in the technique of vascular perfusion-fixation employing a fluorocarbon-containing perfusate and a peristaltic pump controlled by pressure feedback
AU - Rostgaard, J
AU - Qvortrup, Klaus
AU - Poulsen, Steen Seier
PY - 1993/11
Y1 - 1993/11
N2 - A new improved technique for whole-body perfusion-fixation of rats and other small animals is described. The driving force is a peristaltic pump which is feedback regulated by a pressure transducer that monitors the blood-perfusion pressure in the left ventricle of the heart. The primary perfusate-fixative is composed of a blood substitute--13.3% oxygenated fluorocarbon FC-75--in 0.05 M cacodylate buffer (pH 7.4) with a 2% glutaraldehyde. The secondary perfusate-fixative is composed of 2% glutaraldehyde in 0.05 M cacodylate buffer (pH 7.4) with 20 mM CaCl2. A double-barrelled, self-holding cannula is used to cannulate the heart; the outer and inner barrels of the cannula are connected to the peristaltic pump and to the pressure transducer, respectively. The tissue oxygen tension in the rat is monitored by a subcutaneous oxygen electrode. Measurements showed that tissue hypoxia/anoxia did not develop before or during the perfusion-fixation. Thus, the technique permits study of specimens which do not exhibit fixation gradients and do not contain cells fixed in a state of asphyxia. This is substantiated by electron micrographs of cells from different organs, revealing new fine structural elements. By adding oxygenated fluorocarbon to glutaraldehyde perfusate-fixatives, enough oxygen is made accessible for cellular respiration as well as for the oxygen-consuming chemical reactions of glutaraldehyde with the tissue. Data on anaesthesia, operative manoeuvres, mechanical components of the system, preparation of fixatives and flow of the perfusate-fixatives are furnished and discussed.
AB - A new improved technique for whole-body perfusion-fixation of rats and other small animals is described. The driving force is a peristaltic pump which is feedback regulated by a pressure transducer that monitors the blood-perfusion pressure in the left ventricle of the heart. The primary perfusate-fixative is composed of a blood substitute--13.3% oxygenated fluorocarbon FC-75--in 0.05 M cacodylate buffer (pH 7.4) with a 2% glutaraldehyde. The secondary perfusate-fixative is composed of 2% glutaraldehyde in 0.05 M cacodylate buffer (pH 7.4) with 20 mM CaCl2. A double-barrelled, self-holding cannula is used to cannulate the heart; the outer and inner barrels of the cannula are connected to the peristaltic pump and to the pressure transducer, respectively. The tissue oxygen tension in the rat is monitored by a subcutaneous oxygen electrode. Measurements showed that tissue hypoxia/anoxia did not develop before or during the perfusion-fixation. Thus, the technique permits study of specimens which do not exhibit fixation gradients and do not contain cells fixed in a state of asphyxia. This is substantiated by electron micrographs of cells from different organs, revealing new fine structural elements. By adding oxygenated fluorocarbon to glutaraldehyde perfusate-fixatives, enough oxygen is made accessible for cellular respiration as well as for the oxygen-consuming chemical reactions of glutaraldehyde with the tissue. Data on anaesthesia, operative manoeuvres, mechanical components of the system, preparation of fixatives and flow of the perfusate-fixatives are furnished and discussed.
KW - Animals
KW - Blood Circulation
KW - Blood Pressure
KW - Blood Substitutes
KW - Equipment Design
KW - Fluorocarbon Polymers
KW - Fluorocarbons
KW - Glutaral
KW - Guinea Pigs
KW - Male
KW - Mice
KW - Perfusion
KW - Poloxalene
KW - Rabbits
KW - Rats
KW - Rats, Wistar
KW - Tissue Fixation
M3 - Journal article
C2 - 8289232
VL - 172
SP - 137
EP - 151
JO - Journal of Microscopy
JF - Journal of Microscopy
SN - 0022-2720
IS - Pt 2
ER -
ID: 47487398