Conservation des Lipides par les Techniques Utilis�es en Microscopie �lectronique

Idelman, S

doi:10.1007/bf00307888

Cited by 22 publications

(4 citation statements)

References 8 publications

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“…1 The possibility that PA-Schiff-reactive glycolipids are involved in the Golgi apparatus has to be considered, since there are indications that lipids occur in this organelle (59). However, there is good evidence that lipids are almost completely extracted from glutaraldehyde-fixed tissues by the absolute alcohol and propylene oxide used in processing (60,61). FIGURES 14-16 Glandular epithelia.…”

Section: Validity Of the Techniquementioning

confidence: 99%

Detection of Complex Carbohydrates in the Golgi Apparatus of Rat Cells

Rambourg

Hernandez

Leblond

1969

The Journal of Cell Biology

334

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Two methods used for the electron microscopic detection of glycoproteins were applied to a variety of cell types in the rat; one involved successive treatment of sections with periodic acid, chromic acid; and silver methenamine; and the other, a brief treatment with a chromic acid-phosphotungstic acid mixture. The results obtained with the two methods were identical and, whenever the comparison was possible, similar to those obtained with the periodic acid-Schiff technique of light microscopy. In secretory as well as in nonsecretory cells, parts of the Golgi apparatus are stained. The last saccule on one side of each Golgi stack is strongly reactive (mature face), and the last saccule on the other side shows little or no reactivity (immature face); a gradient of reactivity occurs in between these saccules. The more likely explanation of the increase in staining intensity is that carbohydrate is synthesized and accumulates in saccules as they migrate toward the mature face. In many secretory cells, the mature face is associated with strongly stained secretory granules. Other structures stained are: (1) small vesicles, dense and multivesicular bodies, at least some of which are presumed to be lysosomal in nature; (2) cell coat; and (3) basement membrane. The evidence suggests that the Golgi saccules provide glycoproteins not only for secretion, but also for the needs of the lysosomal system as well as for incorporation into the cell coat and perhaps basement membrane.

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Section: Validity Of the Techniquementioning

confidence: 99%

Detection of Complex Carbohydrates in the Golgi Apparatus of Rat Cells

Rambourg

Hernandez

Leblond

1969

The Journal of Cell Biology

334

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“…During the shortened ethanolic dehydration used in the Epon schedule (Idelman, 1965) 33.7% of the total label recovered was extracted from the tissue in ethanol even though the absolute alcohol stage was omitted. This confirms the unsuitability of ethanolic dehydration in cytochemical studies of lipids by EM.…”

Section: Discussionmentioning

confidence: 99%

“…Aldehydes, such as glutaraldehyde, are efficient cross-linking agents for proteins but seem to have little effect on most lipids (Carlier et al, 1968;Idelman, 1965). As much as 95% of total lipid can be lost during ethanolic dehydration following fixation with glutaraldehyde alone (Korn & Weisman, 1966).…”

Section: Introductionmentioning

confidence: 99%

“…None of the embedding procedures described has resulted in the total quantitative preservation of neutral lipid and special processing schedules have been suggested to overcome this problem. Procedures using ethanolic dehydration seem generally best avoided if lipids are to be examined although Idelman (1965) suggested the use of Epon embedding omitting both the 1000/, ethanol and propylene oxide stage. Stein & Stein (1968) reported a 78O/, retention of total lipid using this method.…”

Section: Introductionmentioning

confidence: 99%

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Lipid losses during processing of cardiac muscle for electron microscopy

Ward

Gloster

1976

Journal of Microscopy

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SUMMARY The lipid retaining properties of several methods of processing tissue for electron microscopy (EM) have been assessed quantitatively. Guinea‐pig hearts were perfused in vitro at 37°C with 3H‐oleic acid bound to albumin. The hearts were fixed by perfusion with 4% glutaraldehyde in 0.1 M cacodylate buffer. Pieces of left ventricle and interventricular septum were removed., weighed and processed for EM. The fluids used at each stage of processing were monitored for loss of radioactive lipid by scintillation counting. Lipids were extracted from the processed tissue immediately before the embedding stage using a mixture of chloroform: methanol (2:1 v/v). Counts from processed tissue were compared with counts from tissue extracted directly after perfusion fixation in order to monitor subsequent losses during processing. A modified version of Epon processing, omitting 100% ethanol, acetone or propylene oxide, gave a lipid retention of only 20.6%. The addition of para‐phenylenediamine to the procedure did not improve the retention although this has been shown to be a useful stain for intracellular lipid. Water soluble Durcupan which does not involve ethanol or acetone dehydration has an average retention of 63% with 100% recovery while the lesser known polymer GACH, a mixture of glutaraldehyde and carbohydrazide used both for dehydration and embedding showed a lipid retention of 82% of the counts recovered although recovery was only 69%. An attempt was made to determine which classes of lipids were present in the tissue after perfusion fixation using thin layer chromatography. It was found that the presence of any of the processing fluids affected the polarity of the lipids and their rates of migration on thin layer plates.

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The demonstration of phospholipids in certain cells in neurosecretory nuclei in the rat with Baker's acid haematein after fixation in glutaraldehyde-formol-calcium

Kroon

Goossens

1967

Z. Zellforsch.

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Conservation des Lipides par les Techniques Utilis�es en Microscopie �lectronique

Cited by 22 publications

References 8 publications

Detection of Complex Carbohydrates in the Golgi Apparatus of Rat Cells

Detection of Complex Carbohydrates in the Golgi Apparatus of Rat Cells

Lipid losses during processing of cardiac muscle for electron microscopy

The demonstration of phospholipids in certain cells in neurosecretory nuclei in the rat with Baker's acid haematein after fixation in glutaraldehyde-formol-calcium

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