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Briefly, fixed samples were washed in 0.1 M cacodylate solution, post-fixed with osmium tetroxide, treated with tannic acid, dehydrated with ethanol and then dried in a CPD 030 critical point dryer.
One hour later, the lung was collected and then briefly fixed in 3.7% formaldehyde for 30 min on ice, and then embedded into the Tissue-Tek O.C.T compound followed by cryosections.
Here we show both qualitatively and quantitatively that compared to air-dried and briefly fixed parasites, longer fixation in suspension leads to improved conservation of nuclear structure and lower intra-population variation of nuclear shape as well as area after FISH development.
Cells were briefly fixed in 4% paraformaldehyde and then permeabilized with 0.5% Triton-X in PBS.
For X-Gal staining embryos were collected at desired stages (9.5 10.5 dpc) and briefly fixed in 4% PFA.
Briefly, fixed tissue samples were rinsed in running tap water overnight, dehydrated in a graded series of ethanols, cleared, embedded in Paraplast (Leica Microsystems, Wetzlar, Germany), sectioned at 4 µm and stained with hematoxylin and eosin following standard histological techniques.
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GUS staining was performed by briefly fixing tissue in 90% acetone and then incubating tissue overnight in a 50 mM phosphate buffer containing X-Gluc and a mixture of potassium ferricyanide and ferrocyanide at 37°C.
Briefly, a fixed amount of the radiolabeled antibody was incubated with an increasing number of SKRC38 cells expressing CAIX.
Briefly, formalin fixed paraffin-embedded kidney sections (4 μm) were cut and mounted on gelatinized slides.
Briefly, the fixed tissue fragments were dehydrated, cleared, and embedded in paraffin.
Briefly, formaldehyde fixed, paraffin-embedded tissue sections (5 μm) were deparaffinized and rehydrated in xylene and degradation alcohol.
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