For both compression and suspension, the weakest fixation was attained with cancellous fixation devices.
Approaches include vaginal, e.g. uterosacral ligament suspension, sacrospinous ligament fixation, open procedures and more recently laparoscopic, e.g. sacrocolpopexy and uterosacral plication [2, 5].
Adherent cells have to be detached into a single cell suspension prior to fixation.
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.
The bovine sperm cells were prepared by the institute "Lazzaro Spallanzani" after fixation in suspension of the seminal material with 0.2% glutaraldehyde solution in phosphate buffered saline (PBS) without calcium and magnesium (1 3 v/v).
Phage infections were performed as described above for the colony protocol, but after fixation cell suspensions were kept at 4°C until use, no washing step was necessary before injection of the samples in the flow cytometer.
Cells were trypsinized and hypotonized in 38 mM KCl for 5 min. After fixation, cell suspensions were stored overnight at −20°C.
Other procedures performed simultaneously included sacral colpopexy, sacrospinous suspension, rectopubic urethropexy, and abdominal fixation of the vagina to the uterosacral ligaments.
After membrane staining, erythrocyte lysis, and leukocytes fixation, the cell suspension was permeabilized with PBS-0.05% BSA buffer supplemented with 0.5% saponin.
