Improved section adhesion for immunocytochemistry using high molecular weight polymers of l-lysine as a slide coating

Huang, Wei Min; Gibson, Susan E.; Facer, P.; Gu, Jiang; Polak, Julia M.

doi:10.1007/bf00506570

Cited by 329 publications

(86 citation statements)

References 6 publications

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“…Precleaned glass slides were coated with an aqueous solution of 0.05% (w/v) poly-L-lysine (approximate Mr 300000; Huang et al, 1983). Drops of protoplast suspension were applied to the coated glass slides, incubated for 5 min and protoplasts were fixed by immersion of the slides in 96% ethanol for 20 min.…”

Section: Methodsmentioning

confidence: 99%

Tubular structures involved in movement of cowpea mosaic virus are also formed in infected cowpea protoplasts

Lent¹,

Storms²,

Meer³

et al. 1991

Journal of General Virology

167

104

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In cowpea plant cells infected with cowpea mosaic virus, tubular structures containing virus particles are formed in the plasmodesmata between adjacent cells; these structures are supposedly involved in cell-to-cell spread of the virus. Here we show that similar tubular structures are also formed in cowpea protoplasts, from which the cell wall and plasmodesmata are absent. Between 12 and 21 h post-inoculation, tubule formation starts in the periphery of the protoplast at the level of the plasma membrane. Upon assembly, the viruscontaining tubule is enveloped by the plasma membrane and extends into the culture medium. This suggests that the tubule has functional polarity and makes it likely that a tubule 'grows' into a neighbouring cell in vivo. On average, 75 % of infected protoplasts were shown to possess tubular structures extending from their surface. The tubule wall was 3 to 4 nm thick and they were up to 20 gm in length, as shown by fluorescent light microscopy and negative staining electron microscopy. By analogy to infected plant cells, both the viral 58K/48K movement and capsid proteins were located in these tubules, as determined by immunofluorescent staining and immunogold labelling using specific antisera against these proteins. These results demonstrate that the formation of tubules is not necessarily dependent on the presence of plasmodesmata or the cell wall, and that they are composed, at least in part, of virus-encoded components.

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Section: Methodsmentioning

confidence: 99%

Tubular structures involved in movement of cowpea mosaic virus are also formed in infected cowpea protoplasts

Lent¹,

Storms²,

Meer³

et al. 1991

Journal of General Virology

167

104

View full text Add to dashboard Cite

show abstract

“…For IGSS, the sections were fixed overnight (2.5% glutaraldehyde in PBS), washed in PBS, dehydrated in ethanol, incubated in xylol and paraplast, cut to 6-m sections, and placed on poly-L-lysine-coated glass coverslips. Thereafter the paraplast was removed, the specimens were rehydrated, rinsed in distilled water and PBS (32), incubated in diluted (1:10) normal rabbit serum, then placed for 1 h in the primary polyclonal rabbit anti-ICln-antibody-containing solution (14), washed and incubated with the labeled (10-nm gold) secondary anti-rabbit IgG antibody. Then the samples were extensively washed, incubated in the dark for 5-10 min in a 1:1 mixture of hydrochinon and silver acetate (British Bio Cell International), washed extensively again in distilled water, subjected to iron hematoxilin staining and evaluated on a confocal laser-scanning microscope (LSM 410, Zeiss).…”

Section: Immunohistochemistry/immunogold Silver Staining (Igss)mentioning

confidence: 99%

Cell Swelling Stimulates Cytosol to Membrane Transposition of ICln

Ritter

Ravasio

Jakab

et al. 2003

Journal of Biological Chemistry

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ICln is a multifunctional protein that is essential for cell volume regulation. It can be found in the cytosol and is associated with the cell membrane. Besides its role in the splicing process, ICln is critically involved in the generation of ion currents activated during regulatory volume decrease after cell swelling (RVDC). If reconstituted in artificial bilayers, ICln can form ion channels with biophysical properties related to RVDC. We investigated (

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“…Tumour blocks were sectioned (5yu) on a microtome and sections layered onto clean, sterile, poly-L-lysine coated glass slides (Huang et al, 1983). These were baked at 37°C overnight prior to storage at room temperature in a dry, dust-free box.…”

Section: In Situ Hybridisationmentioning

confidence: 99%

In situ hybridisation and S1 mapping show that the presence of infiltrating plasma cells is associated with poor prognosis in breast cancer

Parkes

Collis²,

Baildam³

et al. 1988

Br J Cancer

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Summary In order to identify potential markers of prognosis in breast cancer, representative cDNA libraries were constructed using RNA isolated from primary breast tumour tissue associated with good and poor prognosis. Cross-screening of these libraries repeatedly identified cloned mRNA species associated with the immune system, in particular B-cells, in libraries derived from tumours of poor prognosis. We have used one of these, a KIV light chain cDNA probe, in two complementary studies to investigate the relationship between immunoglobin gene expression and prognosis. The results obtained using a combination of S, mapping, RNA blotting and in situ hybridisation demonstrate that the presence of plasma cells, as defined by infiltrating cells which express high levels of immunoglobulin K-chain mRNA, is associated with a poor prognosis.

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Improved section adhesion for immunocytochemistry using high molecular weight polymers of l-lysine as a slide coating

Cited by 329 publications

References 6 publications

Tubular structures involved in movement of cowpea mosaic virus are also formed in infected cowpea protoplasts

Tubular structures involved in movement of cowpea mosaic virus are also formed in infected cowpea protoplasts

Cell Swelling Stimulates Cytosol to Membrane Transposition of ICln

In situ hybridisation and S1 mapping show that the presence of infiltrating plasma cells is associated with poor prognosis in breast cancer

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