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https://www.selleckchem.com/products/vx-561.html Ovarian follicle growth and oocyte maturation depend on the viability of granulosa cells (GC). We quantified GC in whole mouse follicles. Single follicles were isolated from adult mouse ovaries and stained with DAPI or Live-Dead stain before fixation. An objective image analysis protocol for counting fluorescent labeled GC was developed that used Image J software to measure GC cytoplasmic and nuclear areas. These data were compared to the number of GC obtained by disaggregating 96 follicles with enzymes to produce a suspension of GC, which then was stained with trypan blue and assessed using a hemocytometer. We found a linear relation between GC/follicle and follicle diameter. Viability of GC/follicle ranged from 40 ± 11 to 72 ± 7%. The coefficient of variation for image analysis of DAPI stained GC by different assessors was 4%, but the number of GC obtained from image analysis was approximately 50% less than from disaggregated follicles. The number of GC in intact mouse follicles was greater than the number reported earlier for fixed ovarian sections. We found that the number of GC was less in fluorescence labeled follicles; it is possible that the three-dimensional structure of the intact follicles obscured the fluorescent signal. Direct quantification of viable GC isolated from follicles appears to be the most accurate method.BACKGROUND Chronic pain after moderate-to-severe traumatic brain injury (TBI) is associated with notable sensory alterations. Although the incidence of TBI is rapidly growing in older populations, elderly individuals have been largely excluded from sensory testing studies, thus limiting evidence regarding the influence of age on pain-related sensory alterations after TBI. This study aimed to investigate the effect of age on the sensory profiles of patients with and without chronic pain after moderate-to-severe TBI. METHODS Thermal and mechanical quantitative sensory testing were performed on t

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