This is a preprint, a preliminary version of a manuscript that has not completed peer review at a journal. Research Square does not conduct peer review prior to posting preprints. The posting of a preprint on this server should not be interpreted as an endorsement of its validity or suitability for dissemination as established information or for guiding clinical practice.
Preparation of cultured cells for scanning electron microscope
Carol Heckman, Sucheta Kanagasundaram, Marilyn Cayer, Jong Paige
Table 1. Major protrusions are affected by temperature difference between sample and fixative. THP1 monocytic cells were adhered on fibrinogen matrix and primed with 100 ng/mL of GM-CSF overnight at 37 degrees C. Further information on the THP1 cells can be found elsewhere (Kanagasundaram, submitted for publication). The cells were fixed with 3% formaldehyde made fresh from paraformaldehyde in PBS. Sample A was equilibrated to room temperature and exposed to fixative solution that was also at room temperature. Sample B was also exposed to fixative solution at room temperature but the cells were at 37 degrees C just before fixation. In sample C, both the sample and fixative were at 37 degrees C. Actin staining was done by introducing TRITC-conjugated phalloidin. The cells were imaged by confocal microscopy. The major protrusions were measured as described in reference 3.