Embedding cells

Fig 5 Immunogold labelling on thin sections of low-temperature embedded cell walls from 9-day old tobacco cells with JIM 5, a monoclonal antibody that recognises a relatively unesterified pectic epitope. Cell walls of elongating cells label very weakly, but material that is being secreted into the culture medium labels strongly. The old part of the wall is labelled but new wall material is not. 

Crockett DK, Lin Z, Vaughn CP, et al. Identification of proteins from formalin-fixed paraffin-embedded cells by LC-MS/MS. Lab. Invest. 2005 85 1405-1415. 

Compound (Miles Laboratories, Elkhart, IN), snap-frozen, and cut into sections for comparison with paraffin-embedded cell sections (3) FFPE Cell Blocks Six cell pellets were fixed in 10% neutral buffered formalin immediately after harvest, at room temperature for 6,12,24h, 3,7, and 30 days, respectively. For further comparison with the cell model system, recently collected sample of human breast cancer tissues were processed by OCT-embedding and snap-freezing the corresponding routine FFPE block that was obtained from the Norris Cancer Hospital and Research Institute at the University of Southern California Keck School of Medicine (USC). This tissue block was processed routinely (formalin-fixed 24h and processed by automatic equipment). 

For the frozen cell/tissue samples, RNA extraction was carried out by using the TRIzol reagent kit. For the paraffin-embedded cell/tissue, RNA extraction was carried out by two methods heating and nonheating using enzyme digestion for comparison. RT-PCR was performed to compare the results. A... 

Krogerus L, Anderson L. A simple method for the preparation of paraffin-embedded cell blocks from fine needle aspirates, effusions and brushings. Acta Cytol. 1988 32 585-587. 

Fig. 1. Diagram of an EM immunogold assay localizing a protein on plastic sections. The primary antibody binds to an exposed surface epitope of the embedded cells. The antibody is then visualized by binding a second antibody coupled to a colloidal gold particle. The electron-dense gold particle visibly marks the position of the bound antibodies when visualized with the electron microscope. 

In a similar way we can prove that the embedded cell complex of the molecular (4,2)-torus link (see Figure 18) is topologically chiral. Also, by adding appropriate labels we can similarly prove the topological chirality of the oriented embedded cell complex of the molecular Hopf link (see Figure 19). 

Fartasch M, Goerdt S, Hornstein OP. Possibilities and limits of paraffin-embedded cell markers in diagnosis of primary cutaneous histiocytosis. Hautarzt. 1995 46 144-153. 

Soule N, Mui K, O Brien M, et al. A rapid automated system for creating paraffin-embedded cell blocks. Abstract 193. Cancer Cytopathol. 2006 108(Suppl 5) 444-445. 

Effusion samples Immunocytochemical stains should be performed on formalin-fixed, paraffin embedded cell block sections. 

The primary components of LCM technology are (1) visualization of the cells of interest through microscopy, (2) transfer of near-infrared laser energy pulses to a thermolabile polymer with formation of a polymer-cell composite, and (3) removal of the polymer from the tissue surface, which shears the embedded cells of interest away from the heterogeneous tissue section (18,19). Extraction buffers applied to the polymer film solubilize the cells, liberating the molecules of interest. The DNA, RNA, or protein from the microdissected cells may be analyzed by any method with appropriate sensitivity (20,21,22,23,24). Protein extracted from microdissected cells may be used for mass spectrometric analysis, applied to reverse phase protein microarrays, or used for western blot analysis (25,26). 

Insert or delete rows as appropriate anywhere except at the top or bottom rows of each block to avoid cancellation of embedded cell formnlas. 

Capsules provide viable enclaves for human cell proliferation, growth and differentiation. All skeletal cell populations remain viable at 91% (+/— 6%) within capsules at differing concentrations of calcium (5-25 mM) and phosphate ions (50-300 mM) used to vary shell thickness and hence diffusive and mechanical properties of the capsule. From histological observations proliferation is initially slow up until 21 days. Embedded cells remain rounded-up but retain their phenotype. Cell-cell interactions stimulate the formation of cell contacts and re-shaping of cell morphology. 

Figure 3 Double labelling of a- and p-tubulin components of microtubules labelled with 5 nm and 10 nm gold conjugates in a section of a PLT embedded cell. Whilst lines of >ld marker can be seen occasionally following the profile of a microtubule (arrow) this figure illustrates the point that although the microtubules can be seen in the section, only those antigens accessible at the section surface can be labelled. Magnification x 74000.

Embedding cells in agarose on microscope slides 427 Staining embedded DNA 428... 

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