Microperfusion fixation of embryos for ultrastructural studies

Abrunhosa, Rui

doi:10.1016/s0022-5320(72)90045-7

Cited by 27 publications

(5 citation statements)

References 21 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…The choice of fixation protocol will need to strike a balance between the degree of neuro-architectural preservation and practical considerations such as technical skill required and speed. In general, nervous tissue is preserved best by perfusion fixation, and the protocols for this procedure can be utilized readily in older embryos and neonates (McKerlie, Newbigging, and Wood 2015) and with practice in embryos (Abrunhosa 1972). Common fixatives for perfusion techniques are aldehydes, such as 10% neutral buffered formalin (NBF, which in commercial solutions typically contains methanol as a stabilizer to inhibit the oxidation of formaldehyde to formic acid); 4% paraformaldehyde (PFA 1 ) and, if electron microscopy is of interest, modified Karnovsky’s fixative (2.5% glutaraldehyde with 2% PFA, or similar mixtures).…”

Section: Methodsmentioning

confidence: 99%

Histology Atlas of the Developing Prenatal and Postnatal Mouse Central Nervous System, with Emphasis on Prenatal Days E7.5 to E18.5

et al. 2017

View full text Add to dashboard Cite

Evaluation of the central nervous system (CNS) in the developing mouse presents unique challenges given the complexity of ontogenesis, marked structural reorganization over very short distances in three dimensions each hour, and numerous developmental events susceptible to genetic and environmental influences. Developmental defects affecting the brain and spinal cord arise frequently both in utero and perinatally as spontaneous events, following teratogen exposure, and as sequelae to induced mutations, and thus are a common factor in embryonic and perinatal lethality in many mouse models. Knowledge of normal organ and cellular architecture and differentiation throughout the mouse’s lifespan is crucial to identify and characterize neurodevelopmental lesions. By providing a well-illustrated overview summarizing major events of normal in utero and perinatal mouse CNS development with examples of common developmental abnormalities, this annotated, color atlas can be used to identify normal structure and histology when phenotyping genetically engineered mice (GEM) and will enhance efforts to describe and interpret brain and spinal cord malformations as causes of mouse embryonic and perinatal lethal phenotypes. The schematics and images in this atlas illustrate major developmental events during gestation from embryonic day (E) 7.5 to E18.5 and after birth from postnatal day (P)1 to P21.

show abstract

Section: Methodsmentioning

confidence: 99%

Histology Atlas of the Developing Prenatal and Postnatal Mouse Central Nervous System, with Emphasis on Prenatal Days E7.5 to E18.5

et al. 2017

View full text Add to dashboard Cite

show abstract

“…The fetuses were distributed as follows: day 9, three; day 10, three; day 11, four; day 11%, one; day 12, nine; day 12% three; day 13, nine; day 13%, four; day 14, eight; day 15, eight; day 16, six; day 17, four; day 18, three; and day 19, three. The fetuses were fixed in situ by intracardiac perfusion (Abrunhosa, 1972) with 2.5% glutaraldehyde in 0.1 M cacodylate buffer, postfixed in 1% buffered Os04, and then embedded in Epon 812.…”

Section: Methodsmentioning

confidence: 99%

Ephemeral, rudimentary glomerular structures in the mesonephros of the mouse

Zamboni

Upadhyay

1981

Anat. Rec.

View full text Add to dashboard Cite

The fine morphology of glomeruli present in the mesonephroi of mouse embryos from days 12 to 14 of gestational age is described. The glomeruli are rudimentary, ephemeral structures which probably result from temporary juxtaposition of a blood capillary to the ventral extremity of a mesonephrictubule. Yet, they might possess a certain degree of functionality.While the studies of Leeson (1960) in the rabbit, Davies and Davies (1950) and Zamboni et al. (1981) in the sheep, and De Martino and Zamboni (1966) in the human have shown that the mesonephros in these species has a complex structural organization, the transient kidney of rodents has been said to consist only of tubules and to lack identifiable glomeruli; as such, it is considered to be rudimentary and nonfunctional (Bremer, 1916;Brambell, 1927;Brown, 1931;Torrey, 1943).In a more recent investigation, Vetter and Gibley (1966) demonstrated instead that the mouse mesonephros possesses two types of rudimentary glomeruli external, seen as clusters of epithelial cells protruding into the coelomic cavity from the ventral surface of the organ, and internal, more complex structures in association with mesonephric tubules and blood vessels deriving from the aorta. These authors, thus, concluded that the mesonephros of the mouse is provided with a certain degree of functionality. However, due to the fact that their study was conducted either on paraffin sections stained with hematoxylin and eosin or by histochemical procedures, the report of Vetter and Gibley (1966) failed to elucidate ade quately the structural organization of the glomeruli, an important point if the function of the mouse mesonephros is to be assessed with a reasonable degree of accuracy.In the course of a study on the role played by the mesonephros in the development of the mouse gonads, we were able to observe the occurrence of glomeruli and to study their structural organization to a degree of resolution higher than that with which these structures were described in previous reports. MATERIALS AND METHODSThe mesonephroi of 68 mouse fetuses collected at close intervals of development, from day 9 after vaginal plug detection to term (day 19), were studied. The fetuses were distributed as follows: day 9, three; day 10, three; day 11, four; day 11%, one; day 12, nine; day 12% three; day 13, nine; day 13%, four; day 14, eight; day 15, eight; day 16, six; day 17, four; day 18, three; and day 19, three. The fetuses were fixed in situ by intracardiac perfusion (Abrunhosa, 1972) with 2.5% glutaraldehyde in 0.1 M cacodylate buffer, postfixed in 1% buffered Os04, and then embedded in Epon 812.The tissue was serially sectioned for light microscopy at i-pm thickness, and the sections were stained in 1% aqueous Toluidine blue. Upon reaching areas of interest, the blocks were trimmed and thin-sectioned for electron microscopy . RESULTSIn the mouse, mesonephric glomeruli are rare-occurring structures which are present in extremely limited numbers; they were visualized only in five fetuses-four at day 1 2 a...

show abstract

“…After clearing the vascular bed with isotonic NaCl solution (0.9 %), the organs were perfused with fixative. The fixative was a mixture of formaldehyde-glutaraldehyde (KARNOVSKI, 1965) supplemented with 2 % of polyvinylpyrrolidone (PVP) in 0.1 M cacodylate buffer (pH 7.4) at 5-10°C (ABRUNHOSA, 1972). The perfused ovaries were kept in the fixative at 4°C at least overnight.…”

Section: Methodsmentioning

confidence: 99%

Fine Structure of Corpora Lutea in Superovulated Heifers

Maciel

Rodriguez‐Martinez

Gustafsson

1992

Journal of Veterinary Medicine Series A

View full text Add to dashboard Cite

Summary The aim of the present study was to determine the degree of development and structural status of perfusion‐fixed day‐7 corpora luteae (CL) (using qualitative and quantitative histology at light and electron microscopy levels) in FSH‐induced superovulated (SO) and untreated heifers. Blood samples were collected daily to monitor the plasma progesterone levels during the FSH treatment up to slaughter 7 days after oestrus. Meanwhile the ovarian activity was followed by ultrasonography and rectal palpation. At slaughter, the ovaries were fixed by vascular perfusion to avoid distortion of the structures and conventionally processed for electron microscopy. The volume density of the luteal tissue was calculated on the examined sections by point‐counting. The histology of the corpora lutea, exception made of a higher incidence of degenerated luteal and endothelial cells in the superovulated animals, did not differ from that of the untreated animals, confirming their normal development. The mean weight and volume of the CL in SO animals was 50% smaller than that of the untreated heifers. The progesterone concentration (nmol/l) at day 7 was significantly positively correlated with the number of CLs/heifer (r = 0.93, P < 0.01), with the weight of Cls/heifer (r = 0.97, P < 0.005) and with the volume of CLs/heifer (r = 0.97, P < 0.005). The results indicated that there were no morphological differences, in terms of histological structure and volume density, between the luteal tissue of SO heifers and the luteal tissue of non‐SO heifers on day 7 of the oestrous cycle which could interfere with its endocrine function.

show abstract

Microperfusion fixation of embryos for ultrastructural studies

Cited by 27 publications

References 21 publications

Histology Atlas of the Developing Prenatal and Postnatal Mouse Central Nervous System, with Emphasis on Prenatal Days E7.5 to E18.5

Histology Atlas of the Developing Prenatal and Postnatal Mouse Central Nervous System, with Emphasis on Prenatal Days E7.5 to E18.5

Ephemeral, rudimentary glomerular structures in the mesonephros of the mouse

Fine Structure of Corpora Lutea in Superovulated Heifers

Contact Info

Product

Resources

About