Immunohistochemical Evaluation of the Effects of Paraffin Section Storage on Biomarker Stability
by J. A. Ramos-Vara, J. D. Webster, D. DuSold, and M. A. Miller
Veterinary Pathology, 51(1): 102-109
Abstract:
Environmental stresses can alter immunoreactivity of biomarkers in stored tissue sections. The effect of temperature and lighting
on 49 cellular or microbial antigens was evaluated in 4 serial paraffin sections, cut 12 months, 10 months, 8 months, 5 months, 3
months, 1 month, 3 days, and 1 day before immunohistochemistry. Slides were stored at room temperature (RT) in the dark, at
4C in the dark, at RT under fluorescent light, or at RT with windowpane exposure to sunlight. Immunohistochemistry was
performed simultaneously in an automated immunostainer. Immunoreactivity was compared with that in the corresponding
1-day-old section and scored as 4 (<10% reduction), 3 (10%–25% reduction), 2 (26%–60% reduction), 1(>60% reduction), or 0 (no
reactivity). Any loss of immunoreactivity was proportional to the tissue section age and was least in sections stored in the dark.
Immunoreactivity was only completely lost in light-exposed sections and as early as 1 month for CD45. Other markers with complete
loss of immunoreactivity were bovine viral diarrhea virus, CD18 (only with fluorescent light), CD31, CD68, canine
parvovirus, chromogranins, and thyroid transcription factor-1. Markers with complete loss after light exposure also had reduced
immunoreactivity when stored in the dark, as early as day 3. Eight markers (Bartonella spp, CD11d, high molecular weight cytokeratins,
feline coronavirus, GATA-4, insulin, p63, progesterone receptor) had minimal decrease in immunoreactivity, regardless
of treatment. In conclusion, light-induced antigen decay (tissue section aging) is antigen dependent and could explain unexpectedly
weak or negative immunohistochemical reactions in stored paraffin sections.
by J. A. Ramos-Vara, J. D. Webster, D. DuSold, and M. A. Miller
Veterinary Pathology, 51(1): 102-109
Abstract:
Environmental stresses can alter immunoreactivity of biomarkers in stored tissue sections. The effect of temperature and lighting
on 49 cellular or microbial antigens was evaluated in 4 serial paraffin sections, cut 12 months, 10 months, 8 months, 5 months, 3
months, 1 month, 3 days, and 1 day before immunohistochemistry. Slides were stored at room temperature (RT) in the dark, at
4C in the dark, at RT under fluorescent light, or at RT with windowpane exposure to sunlight. Immunohistochemistry was
performed simultaneously in an automated immunostainer. Immunoreactivity was compared with that in the corresponding
1-day-old section and scored as 4 (<10% reduction), 3 (10%–25% reduction), 2 (26%–60% reduction), 1(>60% reduction), or 0 (no
reactivity). Any loss of immunoreactivity was proportional to the tissue section age and was least in sections stored in the dark.
Immunoreactivity was only completely lost in light-exposed sections and as early as 1 month for CD45. Other markers with complete
loss of immunoreactivity were bovine viral diarrhea virus, CD18 (only with fluorescent light), CD31, CD68, canine
parvovirus, chromogranins, and thyroid transcription factor-1. Markers with complete loss after light exposure also had reduced
immunoreactivity when stored in the dark, as early as day 3. Eight markers (Bartonella spp, CD11d, high molecular weight cytokeratins,
feline coronavirus, GATA-4, insulin, p63, progesterone receptor) had minimal decrease in immunoreactivity, regardless
of treatment. In conclusion, light-induced antigen decay (tissue section aging) is antigen dependent and could explain unexpectedly
weak or negative immunohistochemical reactions in stored paraffin sections.