Samples are prepared by a similar method to transmission electron microscopy, typically by fixing the sample with aldehyde, staining with heavy metals such as osmium and uranium then embedding in an epoxy resin.
Cells or spheres were prepared for scanning electron microscopy (SEM) by fixing with 4% glutaraldehyde in 0.15 m cacodylate buffer, containing 4.7 m NaCl, 80 m m MgSO4 × 7H2O, post-fixing with 1% osmium tetroxide, critical point-drying, and sputter-coating with approximately 2 nm Pt.
Following coculture, free or loosely attached T cells were washed off with 10 mM EDTA, MDMs were cultured, and productive infection was analyzed at day 3 post-coculture by fixing and confocal microscopy.
The homogeneity and the crystal sizes were controlled after matrix application by microscopy before fixing the sample on the sample holder of the imaging source.
Cells used for immunofluorescence microscopy were fixed by 4% formaldehyde and 1 ml of cell suspension were treated by zymolyase T100 (100 μg), glucuronidase (30 μl) and 10 mM DTT for 30 min at 30°C.
The total cell and differential cell counts were determined by microscopy of fixed BAL fluid samples.
TMV-positive RNA and proteins were identified in situ by confocal microscopy in cells fixed by a protocol intended to retain native cell size and shape.
One day after transfection, B16-F1 cells were seeded onto coverslips coated with 25 µg/ml laminin (Sigma-Aldrich) and either examined by video microscopy or fixed and processed for immunolabeling or electron microscopy.
The resulting segmentations are in agreement with data obtained by confocal microscopy in fixed samples using nuclear staining (Ronneberger et al., 2012).
