Influence of storage temperature and high-temperature antigen retrieval buffers on results of immunohistochemical staining in sections stored for long periods
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Influence of storage temperature and high-temperature antigen retrieval buffers on results of immunohistochemical staining in sections stored for long periods. / Grabau, Dorthe A.; Nielsen, Ole; Hansen, Steinbjørn; Nielsen, Mette M.; Lænkholm, Anne Vibeke; Knoop, Ann; Pfeiffer, Per.
In: Applied Immunohistochemistry, Vol. 6, No. 4, 1998, p. 209-213.Research output: Contribution to journal › Journal article › Research › peer-review
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TY - JOUR
T1 - Influence of storage temperature and high-temperature antigen retrieval buffers on results of immunohistochemical staining in sections stored for long periods
AU - Grabau, Dorthe A.
AU - Nielsen, Ole
AU - Hansen, Steinbjørn
AU - Nielsen, Mette M.
AU - Lænkholm, Anne Vibeke
AU - Knoop, Ann
AU - Pfeiffer, Per
PY - 1998
Y1 - 1998
N2 - The aim of this study was to evaluate the importance of storage temperature in paraffin-embedded sections stored for a fixed period of 3 years. Sections were stained with immunohistochemically stained and treated with high-temperature antigen retrieval (AR). If staining results were impaired, an alternative and more efficient AR protocol was compared with the first to show possible restoration of the immunohistochemical staining. The material represented tumor tissue from two lung carcinomas and four breast carcinomas prepared in a multitissue sandwich block. Sections were cut, mounted on glass slides, and stored for 3 years at -80, 4, or 20°C, or unmounted at 4°C. Additional new sections were cut from the original block the day before staining. The immunohistochemical antigens and clones included ER(1D5), Ki67(MIB1), p53(DO7), C-erbB2(poly), RB1(G3.245), bcl2(124), E- cadherin(HECD-1), and EGFR(113). The first set of AR protocols compared citrate buffer and heating for 25 minutes with TEG buffer and heating for 25 minutes. The second set compared heating for 25 versus 35 minutes after TEG buffer. Positive area was quantitated with the Image Analysis CAS 200 system. The results showed that the positive area decreased with increasing storage temperature for all antigens except C-erbB2, regardless of the AR protocol. Storage at 4°C gave some decrease in positive area, but the antigen expression could be restored, except for EGFR, by the use of an alternative and more efficient AR protocol.
AB - The aim of this study was to evaluate the importance of storage temperature in paraffin-embedded sections stored for a fixed period of 3 years. Sections were stained with immunohistochemically stained and treated with high-temperature antigen retrieval (AR). If staining results were impaired, an alternative and more efficient AR protocol was compared with the first to show possible restoration of the immunohistochemical staining. The material represented tumor tissue from two lung carcinomas and four breast carcinomas prepared in a multitissue sandwich block. Sections were cut, mounted on glass slides, and stored for 3 years at -80, 4, or 20°C, or unmounted at 4°C. Additional new sections were cut from the original block the day before staining. The immunohistochemical antigens and clones included ER(1D5), Ki67(MIB1), p53(DO7), C-erbB2(poly), RB1(G3.245), bcl2(124), E- cadherin(HECD-1), and EGFR(113). The first set of AR protocols compared citrate buffer and heating for 25 minutes with TEG buffer and heating for 25 minutes. The second set compared heating for 25 versus 35 minutes after TEG buffer. Positive area was quantitated with the Image Analysis CAS 200 system. The results showed that the positive area decreased with increasing storage temperature for all antigens except C-erbB2, regardless of the AR protocol. Storage at 4°C gave some decrease in positive area, but the antigen expression could be restored, except for EGFR, by the use of an alternative and more efficient AR protocol.
KW - High-temperature antigen retrieval
KW - Immunohistochemistry
KW - Paraffin embedding
KW - Storage time
UR - http://www.scopus.com/inward/record.url?scp=0031786330&partnerID=8YFLogxK
U2 - 10.1097/00022744-199812000-00006
DO - 10.1097/00022744-199812000-00006
M3 - Journal article
AN - SCOPUS:0031786330
VL - 6
SP - 209
EP - 213
JO - Applied Immunohistochemistry
JF - Applied Immunohistochemistry
SN - 1062-3345
IS - 4
ER -
ID: 260459622