PMC

Examination of protein degradation from 20 μm brain tissue sections at defined time points from various storage conditions. Extracted samples were resolved on a denaturing gel and stained by silver method. A, One-day-old samples. B, One-week-old samples. C, One-month-old samples. D, Six-month-old sample. S = Protein calibration standard. 1 = Fresh brain tissue section. 2 = Frozen brain tissue section (without heat). 3 = Ambient brain tissue section (without heat). 4 = Desiccated brain tissue section (without heat). 5 = Frozen brain tissue section (with heat). 6 = Ambient brain tissue section (with heat). 7 = Desiccated brain tissue section (with heat).

Demonstration of epitope detection of target protein extracted from 20 μm brain tissue sections at defined time points from various storage conditions. Samples were resolved on a denaturing gel and Western blot performed for tyrosine hydroxylase (TH). GAPDH served as internal loading control, 1-month samples shown is representative. A, One-day-old sample. B, One-week-old sample. C, One-month-old sample. D, Six-month-old sample. 1 = Fresh brain tissue section. 2 = Frozen brain tissue section (without heat). 3 = Ambient brain tissue section (without heat). 4 = Desiccated brain tissue section (without heat). 5 = Frozen brain tissue section (with heat). 6 = Ambient brain tissue section (with heat). 7 = Desiccated brain tissue section (with heat). GAPDH indicates glyceraldehyde-3-phosphate dehydrogenase.

Comparison of normalized protein concentrations (μg/mL) extracted from 20 μm murine brain tissue sections stored at various conditions over a 6-month period. Frozen refers to tissue sections stored at −80°C, ambient refers to sections stored at room temperature (RT, ~22°C), and desiccated refers to sections stored at RT with low RH. Slides that were not placed on a slide warmer are noted as “wo/h” and those that were are noted as “w/h”. Error bars represent ±SEM. Statistically significantly difference, P<0.05 (Student t test), represented by *. RH indicates relative humidity; w/h, with heat; wo/h, without heat.

Photomicrographs of brain tissue sections with hematoxylin and eosin (H&E), immunoperoxidase, or immunofluorescences stains. Using a rat stereotaxtic atlas as a guide, all samples were taken at approximately bregma 0.72 mm (A←P), at a junction between striatum and cortex. A, C, and E, Fresh brain tissue section dual stain against TH and MAP5B, with nuclear counter stain. B, D, and F, Tissue section from 6-month-old desiccated samples with identical stains. G, Fresh brain GFAP stain. H, GFAP stain of sample after stored in desiccated condition for 6 months. I, Fresh brain H&E stain. J, H&E stain of sample after stored in desiccated condition for 6 months. K, Fresh brain immunoperoxidase stain for TH. L, immunoperoxidase stain for TH of sample after stored in desiccated condition for 6 months. TH = tyrosine hydroxylase, MAP5B = neuron, GFAP = glia (non-neuronal), DAPI = nuclear stain. Photomicrographs were taken at 2 different magnifications, unless otherwise noted, one at 10 × and another at 40 × (shown as inset).