Easy transfer of selected mitoses from light to electron microscopy

Messier, Paul-Emil; Jean, Paul D. De; Richer, Claude‐Lise

doi:10.1159/000132322

Cited by 31 publications

(17 citation statements)

References 4 publications

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“…One cell with a configuration assessed as showing triple synapsis (Fig. 2a) was stained with silver nitrate and transferred to a grid for viewing by electron microscopy (Messier et al, 1986). The trivalent was indeed triply synapsed along its length (Fig.…”

Section: Alpha-satellite Probe L126 (Chromosomes 21 and 13 Centromeres)mentioning

confidence: 99%

Meiotic chromosome pairing in fetal oocytes of trisomy 21 human females

Barlow

Tease

Hultén

2002

Cytogenet Genome Res

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The influence of trisomy on meiotic chromosome association and synapsis was studied in oocytes of two trisomy 21 fetuses. The patterns of association of the three chromosomes 21 were determined by analysis of late zygotene to early diplotene fetal oocytes after immunofluorescent staining of synaptonemal complexes. The identity of chromosome 21 was confirmed using FISH with either a whole chromosome 21 paint or an α-satellite DNA repeat probe. In both fetuses, a wide variety of configurations was present at pachytene. The most common configurations were a trivalent (35.5% and 51.6% of analyzable cells) and a bivalent plus univalent (62.9% and 45.2%). These different frequencies between the fetuses were not significant. Trivalents showed either triple synapsis or double synapsis with pairing-partner switches. The extent of triple synapsis varied from a short segment, either terminal or interstitial, to the whole chromosome length. Through use of immunofluorescent staining of the centromeres, we identified novel types of abnormal chromosome behavior in trisomy 21 fetal oocytes. Thus, we found that 6/41 trivalents had one of the chromosomes associated “out of register,” i.e., in a nonhomologous fashion, with its two homologs. Likewise, we found three cells with bivalent plus univalent configurations, in which the univalent showed self-synapsis. The presence of three copies of chromosome 21 therefore results not only in the formation of complex and highly variable synaptic associations but also causes a significant increase in the occurrence of nonhomologous synapsis in human fetal oocytes.

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Section: Alpha-satellite Probe L126 (Chromosomes 21 and 13 Centromeres)mentioning

confidence: 99%

Meiotic chromosome pairing in fetal oocytes of trisomy 21 human females

Barlow

Tease

Hultén

2002

Cytogenet Genome Res

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“…The transfer of the metaphases to grids was similar to the procedure described by Messier et al (1986) in that the metaphases were first stained or labeled on microscope slides; then the slides were coated, and the metaphases removed with the film were floated off on 0.4% hydrofluoric acid. Instead of Formvar, we used 0.5% polystyrene dissolved in chloroform for coating.…”

Section: Methodsmentioning

confidence: 99%

“…They have been used to investigate G bands , prematurely condensed chromosomes (Schwarzacher et al 1976), silver staining of nucleolus organizer regions (Schwarzacher et al 1978), and silver staining of synaptonemal complexes (Dresser and Moses 1979). The techniques have been modified over the years (Dresser and Moses 1979;Messier et al 1986) and used with different types of EM (Taniguchi and Takayama 1987).…”

Section: Introductionmentioning

confidence: 99%

Demonstration of chromosome replication by BrdU antibody technique and electron microscopy

1990

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We describe the use of different reporter groups in visualizing replication patterns on metaphase chromosomes after BrdU incorporation by the BrdU antibody technique for the electron microscope. There is an inverse correlation between the density of the label and the size of the reporter particles. This observation alludes to stereo problems interfering with the access of the labeled antibodies into the chromatin. The use of silver enhancement enables easy detection of 1-nm and 5-nm gold particles, which make replication patterns visible in the electron microscope as does the diaminobenzidine/H2O2 reaction. Possible consequences for the demonstration of replication patterns and for nonradioactive DNA in situ hybridization are discussed.

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“…1983;Li et al. 1986).Having recently developed a method that permits the easy transfer of any selected mitosis from slide to grid (Messier et al, 1986), we now describe a chromosomal gold-labeling procedure that produces banding specific for EM. …”

mentioning

confidence: 99%

“…The specimens were fixed in 2.5% glutaraldehyde for 30 min. Well-spread mitoses devoid of cytoplasm that were in prometaphase or prophase were selected and transferred from glass slides to grids (Messier et al, 1986). They were photographed, without staining, in a Siemens Elmiskop IA electron microscope operated at 60 kV.…”

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confidence: 99%

Human chromosome banding specific for electron microscopy

Drouin¹,

Messier²,

Richer³

1988

Cytogenet Genome Res

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Electron microscopy (EM) provides much higher resolution than that obtained with light microscopy (LM). Until now, however, no chromosome banding procedure specifically adapted for EM was available. To produce an easy and reproducible banding method that would allow accurate chromosome identification, we investigated the applicability of an immunochemical method. BrdU-substituted chromosomal regions can be accurately visualized by applying a monoclonal antibody against BrdU, followed by a gold-tagged secondary antibody. Since BrdU is incorporated during the last part of the S-phase, regions of darkly stained G- and C-bands are substituted. A characteristic C-banding pattern is revealed, and the G-banding obtained is sharp and allows discrimination between subbands. Its similarity with the classical G-banding observed by LM makes it easy to interpret and facilitates karyotyping.

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Easy transfer of selected mitoses from light to electron microscopy

Abstract: A method is described whereby any given chromosome spread selected by light microscopy can be transferred to a grid and studied by electron microscopy.

Cited by 31 publications

References 4 publications

Meiotic chromosome pairing in fetal oocytes of trisomy 21 human females

Meiotic chromosome pairing in fetal oocytes of trisomy 21 human females

Demonstration of chromosome replication by BrdU antibody technique and electron microscopy

Human chromosome banding specific for electron microscopy

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