Spectrophotometric absorbance of follicular fluid: A selection criterion

Abstract

From this study, it is concluded that blood contamination and dilution with culture medium influence the biochemical composition as well as the absorbance spectrum of follicular fluids. This procedure is advocated as a prerequisite before quantifying FF content.

The variable expression of lectin-like oxidized low-density lipoprotein receptor (LOX-1) and signs of autophagy and apoptosis in freshly harvested human granulosa cells depend on gonadotropin dose, age, and body weight

“…The remaining cell suspension was centrifuged and the pellet treated with protein extraction buffer (80 mmol/L Tris, 70 mmol/L sodium dodecyl sulfate [SDS], and 292 mmol/L saccharose, pH 6,8). The aliquots were stored at À20 C. Before further use, the pellets were screened for the erythrocyte-specific protein hemoglobin (31). The protein-induced absorbance peaked at 418, 540, and 575 nm.…”

Impact of Exogenous Gonadotropin Stimulation on Circulatory and Follicular Fluid Cytokine Profiles

“…Oocyte retrieval frequently results in disruption of the intraovarian vasculature such that blood (macroscopic or microscopic) contaminates the FF retrieved [ 66 ]. Furthermore, although the needle and tubing were primed with normal saline at the commencement of aspiration of each ovary, in between subsequent follicular aspirations, protein-free flush medium formulated with gentamicin (enhanced HTF culture medium with HEPES; Conception Technologies, San Diego, California, USA) was used, with potential dilution of the FF [ 67 ]. Such contamination/dilution was accounted for in the FF cytokine analysis.…”

The detection of blood contamination in human follicular fluid

“…The stimulation protocol and aspiration of hFF have been presented elsewhere (11). Follicular fluids supplemented with heparin or Earle's balanced salt solution medium (spectrophotometric absorbance peak at 562 nrh; Phillips Pye Unicam SP800 spectrophotometer), or fluids that presented with a spectrophotometric peak absorbance at 407--411 nm (cyst fluids), were excluded from this study (21 Once fluids were aspirated and scanned for oocytes, visual inspection was performed by the same skilled person throughout this study, followed by the determination of Hct levels (determined by Hawksley microhematocrit centrifuge, with nonheparinized capillary tubes; Medsurg, RSA).…”

“…Cyst-derived fluids should also be eliminated. Blood and medium contamination leads to altered hFF biochemical profiles with lower steroid concentrations, possibly resulting in observed variations with hFF experimentation (11). The pH of hFF has been shown to decrease significantly with an increasing degree of blood contamination (12).…”

“…Generally applied methods for detecting blood in hFF include visual inspection (14)(15)(16)(17), hematocrit (Hct) (18), and haemoglobin (Hb) determination (19,20), as well as spectrophotometric analysis (11,21). The aim of this study was to compare the reliability of the existing methods for the detection of hFF blood contamination (red blood cells as well as hemoglobin) and to investigate the possibility of using simple alternatives such as visual inspection and the Combur-9-test urine sticks.…”