Easy handling of cell suspensions for electron microscopy

Pihakaski‐Maunsbac, Kaarina; Soitamo, Arto; Suoranta, Ulla‐Maija

doi:10.1002/jemt.1060150412

Cited by 4 publications

(1 citation statement)

References 2 publications

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“…To study the ultrastructure of free or in-suspension cells by means of TEM it is necessary either to centrifuge the samples previously to the fixation, in order to obtain a cellular pellet (Gonzalez Santander, 1970;Krogenaes et al, 1994;Quinn et al, 1969) or to immerse the samples in viscous gels. Among the latter, the most used methods are those in which the samples are preembedded in agarose (Nogues et al, 1994;Watanabe et al, 1988) or those that use the agar encapsulating technique (Pihakaski et al, 1990).…”

Section: Introductionmentioning

confidence: 99%

New method for the treatment of sperm samples for ultrastructural study based on the use of animal tissues as biological containers

Junquera

Colás

Martínez-Ciriano

et al. 2007

Microscopy Res & Technique

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The study of the ultrastructure of spematozoa by means of transmission electron microscopy (TEM) often presents with problems of interpretation according to the method employed, depending on whether samples are either centrifuged previously to the fixation or immersed in viscous gels. The major problems of interpretation are changes in the location of vesicles originated during the maturation process and modifications in the adsorption of seminal plasma proteins to the sperm membrane surface. The aim of our study is to communicate an original new method for the treatment of spermatozoa for ultrastructural study. Our method is based on the use of animal tissues as biological containers, inside which the spermatic suspensions are included. We developed this method using fresh sperm samples taken from mature Rasa Aragonesa rams. As biological container, we used 2.5-cm long segments of the intestine of 1-week-old chickens (Gallus gallus) (diameter around 4 mm). In order to avoid any influence of digestive enzymes of the mucosa on the sperm surface, we put each intestine fragment inside out by means of microdissection forceps under a bifocal optical microscope and cold light. One of the edges was tied with thin suture silk. The sperm suspension was injected in the optimal experimental condition and amount. Finally, the still-open edge of the intestine segment was tied with silk in the same way as the other segment edge. By using this technique, we can perform a suitable morphological study at an ultrastructural level. In addition, the functional relationship of the ultrastructural components of the target cells is correctly preserved.

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Section: Introductionmentioning

confidence: 99%

New method for the treatment of sperm samples for ultrastructural study based on the use of animal tissues as biological containers

Junquera

Colás

Martínez-Ciriano

et al. 2007

Microscopy Res & Technique

View full text Add to dashboard Cite

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A New Method for the Treatment of Sperm Samples for Ultrastructural Study Based on the Use of Animal Tissues as Biological Containers

Junquera

Colás

Martínez-Ciriano

et al. 2007

Microscopy Res & Technique

View full text Add to dashboard Cite

The study of the ultrastructure of spematozoa by means of transmission electron microscopy often presents with problems of interpretation according to the method employed, depending on whether samples are either centrifuged previously to the fixation or immersed in viscous gels. The major problems of interpretation are: changes in the location of vesicles originated during the maturation process and modifications in the adsorption of seminal plasma proteins to the sperm membrane surface. The aim of our study is to communicate an original new method for the treatment of spermatozoa for ultrastructural study. Our method is based on the use of animal tissues as biological containers, inside which the spermatic suspensions are included. We developed this method using fresh sperm samples taken from mature Rasa aragonesa rams. As biological container, we used 2.5-cm long segments of the intestine of 1-week-old chickens (Gallus gallus) (diameter around 4 mm). To avoid any influence of digestive enzymes of the mucosa on the sperm surface, we put each intestine fragment inside out by means of microdissection forceps under bifocal optical microscope and cold light. One of the edges was tied with thin suture silk. The sperm suspension was injected in the optimal experimental condition and amount. Finally, the still open edge of the intestine segment was tied with silk in the same way as the other segment edge. By using this technique, we can perform a suitable morphological study at an ultrastructural level. In addition, the functional relationship of the ultrastructural components of the target cells is correctly preserved.

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Dehydration and Embedding

Maunsbach

Afzelius

1999

Biomedical Electron Microscopy

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Easy handling of cell suspensions for electron microscopy

Abstract: We present here a new, simple agar encapsulating technique, which is helpful when preparing a small quantity of isolated fragile cells, e.g. protoplasts, for electron microscopy.

Cited by 4 publications

References 2 publications

New method for the treatment of sperm samples for ultrastructural study based on the use of animal tissues as biological containers

New method for the treatment of sperm samples for ultrastructural study based on the use of animal tissues as biological containers

A New Method for the Treatment of Sperm Samples for Ultrastructural Study Based on the Use of Animal Tissues as Biological Containers

Dehydration and Embedding

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