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Immunostaining fixation

Figure 16.7 Immunohistochemical staining intensity as a function of the duration of formahn fixation. Immunostaining intensity was measured in triphcate.The coefficients of variation (CVs) were almost aU less than 10% and, for purposes of clarity, are not shown. Staining intensity is measured as mean pixel intensity units on a 1-256 scale. ER(1D5), ER(2-123), PR(636), PR(1294), and Ki-67(MIB-1), DakoCytomation, Carpinteria, CA PR(1A6) and HER2(CB11), Vision Biosystems, NorweU, MA. ER, estrogen receptor PR, progesterone receptor. Reproduced with permission from Reference 15, 2006 American Society for Clinical Pathology. Figure 16.7 Immunohistochemical staining intensity as a function of the duration of formahn fixation. Immunostaining intensity was measured in triphcate.The coefficients of variation (CVs) were almost aU less than 10% and, for purposes of clarity, are not shown. Staining intensity is measured as mean pixel intensity units on a 1-256 scale. ER(1D5), ER(2-123), PR(636), PR(1294), and Ki-67(MIB-1), DakoCytomation, Carpinteria, CA PR(1A6) and HER2(CB11), Vision Biosystems, NorweU, MA. ER, estrogen receptor PR, progesterone receptor. Reproduced with permission from Reference 15, 2006 American Society for Clinical Pathology.
Below are four fixation/immunostaining protocols. Each of these protocols proved to be suitable for the immunolocalization of one or more proteins along mitotic chromosomes. [Pg.38]

Shidham VB, Chang C-C, Rao RN, et al. Immunostaining of cytology smears a comparative study to identify the most suitable method of smear preparation and fixation with reference to commonly used immunomarkers. Diagn. Cytopathol. 2003 29 217-221. [Pg.41]

Standardization of IHC/ICC has been a critical issue for more than three decades, especially with the advances in targeted therapy such as the development of trastuzumab (Herceptin) for advanced breast cancer.51 Nevertheless, standardization is a difficult issue because numerous factors may influence the consistency and reliability of immunostaining results, including fixatives, fixation time, AR, antibody clones, detection system, and interpretation (see Part II). In cytopathology, the situation is even worse due to its variable cell sample preparation techniques. Cytopreparation is. .. the foundation of cytomorphology. 52 We believe it is also the foundation of ICC. Therefore, standardization of ICC needs to start with uniform and reliable cytopreparation. [Pg.228]

The far left column of Figure 16.1 shows a scanned image of the duplicate immunostained peptide spots before formalin fixation. The fact that the spots are stained demonstrates that each of the peptides (p53, FIER2, ER, PR) binds... [Pg.289]

Figure 16.5 Immunostained peptide arrays after various treatments of fixation, protein cross-linking, and antigen retrieval, as indicated at the top. Each row has a different peptide that is immunoreactive for the antibody denoted to the left. Column A represents the baseline condition, without any treatment whatsoever. Column B shows immunoreactivity of each peptide after overnight formalin fixation. Column C shows the immunoreactivity after first coating the array with an irrelevant protein (casein) followed by overnight formalin fixation. Column D illustrates the immunoreactivity of the peptides after the treatment of column C, and then antigen retrieval. Reproduced with permission from Reference 15, 2006 American Society for Clinical Pathology. Figure 16.5 Immunostained peptide arrays after various treatments of fixation, protein cross-linking, and antigen retrieval, as indicated at the top. Each row has a different peptide that is immunoreactive for the antibody denoted to the left. Column A represents the baseline condition, without any treatment whatsoever. Column B shows immunoreactivity of each peptide after overnight formalin fixation. Column C shows the immunoreactivity after first coating the array with an irrelevant protein (casein) followed by overnight formalin fixation. Column D illustrates the immunoreactivity of the peptides after the treatment of column C, and then antigen retrieval. Reproduced with permission from Reference 15, 2006 American Society for Clinical Pathology.
Figure 6. Cytoskeletal proteins in nucleus. HeLa cells were immunostained with anti-keratin 8 or anti-keratin 18 antibodies, (a) Paraformaldehyde-fixed cells, (b) Cells treated with detergent before the fixation. (Scale bars 10pm)... Figure 6. Cytoskeletal proteins in nucleus. HeLa cells were immunostained with anti-keratin 8 or anti-keratin 18 antibodies, (a) Paraformaldehyde-fixed cells, (b) Cells treated with detergent before the fixation. (Scale bars 10pm)...
Battifora, H. and Kopinski, M. (1986) The influence of protease digestion and duration of fixation on the immunostaining of keratins. A comparison of formalin and ethanol fixation. J. Histochem. Cytochem. 34,1095-1100. [Pg.92]

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