Die Diffusion des 59Fe-markierten H�moglobins, ein Artefakt der Glutaraldehyd-Fixierung

Thiessen, G.; Thiessen, H. J.; Dowidat, H. J.; Luciano, L.; Reale, E.

doi:10.1007/bf00305849

Cited by 7 publications

(2 citation statements)

References 2 publications

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“…In addition, we have demonstrated that there is no other source of peroxidatic activity in the plasma, except for a negligible amount of hemoglobin which leaks from erythrocytes, as a result of a slight degree of hemolysis induced by the fixative . The finding is in agreement with observations made by using 59 Fe-labeled hemoglobin (65) . abnormal permeability (vascular leakage) in muscle capillaries .…”

Section: Mb As Probe Moleculesupporting

confidence: 93%

Permeability of Muscle Capillaries to Exogenous Myoglobin

Simiónescu

Simionescu

Palade

1973

The Journal of Cell Biology

185

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Whale skeletal muscle myoglobin (mol wt 17,800 ; molecular dimensions 25 X 34 X 42A) was used as a probe molecule for the pore systems of muscle capillaries . Diaphragms of WistarFurth rats were fixed in situ at intervals up to 4 h after the intravenous injection of the tracer, and myoglobin was localized in the tissue by a peroxidase reaction . Gel filtration of plasma samples proved that myoglobin molecules remained in circulation in native monomeric form . At 30-35 s postinjection, the tracer marked '75% of the plasmalemmal vesicles on the blood front of the endothelium, 157 of those located inside and none of those on the tissue front . At 45 s, the labeling of vesicles in the inner group reached 6070 but remained nil for those on the tissue front . Marked vesicles appeared on the latter past 45 s and their frequency increased to -80% by 60-75 s, concomitantly with the appearance of myoglobin in the pericapillary spaces. Significant regional heterogeneity in initial labeling was found in the different segments of the endothelium (i .e., perinuclear cytoplasm, organelle region, cell periphery, and parajunctional zone) . Up to 60 s, the intercellular junctions and spaces of the endothelium were free of myoglobin reaction product ; thereafter, the latter was detected in the distal part of the intercellular spaces in concentration generally equal to or lower than that prevailing in the adjacent pericapillary space . The findings indicate that myoglobin molecules cross the endothelium of muscle capillaries primarily via plasmalemmal vesicles . Since a molecule of this size is supposed to exit through both pore systems, our results confirm the earlier conclusion that the plasmalemmal vesicles represent the large pore system ; in addition, they suggest that the same structures are, at least in part, the structural equivalent of the small pore system of this type of capillaries . late tracer (1), but the structural identity of the small pore system in the same type of vessels is still in doubt . Using horseradish peroxidase (HRP) (mol wt e-40,000, and -40-50 A) as a histochemically detectable tracer, Karnovsky (2) and Cctran and Karnovsky (3) arrived at the conclusion that it reaches the pericapillary spaces mainly through the intercellular junctions of the 4 2 4

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Section: Mb As Probe Moleculesupporting

confidence: 93%

Permeability of Muscle Capillaries to Exogenous Myoglobin

Simiónescu

Simionescu

Palade

1973

The Journal of Cell Biology

185

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“…This effect is exacerbated when fixing thick tissue pieces because glutaraldehyde diffuses into tissue more slowly than formaldehyde (Reale and Luciano, 1970; Thiessen et al, 1970). This results in a shell of over-fixed tissue on the exterior of the tissue block, and a core of poorly fixed tissue at the center (much like formaldehyde), although various methods for improving antibody penetration after fixation are available (e.g., whole mount electroimmunofluorescence (Liu and Kao, 2009); buffered-ethanol extraction; cryoprotection and subsequent freeze-thaw (Eldred et al, 1983; Voigt and Wässle, 1987).…”

Section: Glutaraldehydementioning

confidence: 99%

Fixation strategies for retinal immunohistochemistry

Stradleigh

Ishida

2015

Progress in Retinal and Eye Research

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Immunohistochemical and ex vivo anatomical studies have provided many glimpses of the variety, distribution, and signaling components of vertebrate retinal neurons. The beauty of numerous images published to date, and the qualitative and quantitative information they provide, indicate that these approaches are fundamentally useful. However, obtaining these images entailed tissue handling and exposure to chemical solutions that differ from normal extracellular fluid in composition, temperature, and osmolarity. Because the differences are large enough to alter intercellular and intracellular signaling in neurons, and because retinae are susceptible to crush, shear, and fray, it is natural to wonder if immunohistochemical and anatomical methods disturb or damage the cells they are designed to examine. Tissue fixation is typically incorporated to guard against this damage and is therefore critically important to the quality and significance of the harvested data. Here, we describe mechanisms of fixation; advantages and disadvantages of using formaldehyde and glutaraldehyde as fixatives during immunohistochemistry; and modifications of widely used protocols that have recently been found to improve cell shape preservation and immunostaining patterns, especially in proximal retinal neurons.

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Theoretical and practical aspects of glutaraldehyde fixation

Hopwood¹

1973

Fixation in Histochemistry

213

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Die Diffusion des 59Fe-markierten H�moglobins, ein Artefakt der Glutaraldehyd-Fixierung

Cited by 7 publications

References 2 publications

Permeability of Muscle Capillaries to Exogenous Myoglobin

Permeability of Muscle Capillaries to Exogenous Myoglobin

Fixation strategies for retinal immunohistochemistry

Theoretical and practical aspects of glutaraldehyde fixation

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