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Impact of formalin fixation on mismatch repair protein evaluation by immunohistochemistry.

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Title: Impact of formalin fixation on mismatch repair protein evaluation by immunohistochemistry.
Authors: Grillo, Federica, Ali, Murad, Paudice, Michele, Pigozzi, Simona, Anselmi, Giorgia, Scabini, Stefano, Sciallero, Stefania, Piol, Nataniele, Mastracci, Luca
Source: Virchows Archiv: European Journal of Pathology; Nov2023, Vol. 483 Issue 5, p677-685, 9p
Abstract: Mismatch repair/microsatellite instability (MMR/MSI) status in colorectal cancer (CRC) has become fundamental as a diagnostic, prognostic, and predictive factor. MMR immunohistochemistry (IHC) is considered a simple and reliable approach; however, its effectiveness depends on pre-analytic factors. Aim of this study was to investigate the impact of different fixation times/protocols on MMR protein IHC quality. Left over tissue from surgically resected CRC samples (cold ischemia time < 30 min) where fixed as follows: standard formalin fixation (24–48 h); hypo-fixation (<20 h); hyper-fixation (>90 h); cold (4°C) fixation (24–48 h); standard fixation for small sample size (0.5×0.5 cm). Samples for each group were collected from 30 resected CRC and the following parameters were evaluated on 600 immunohistochemical stains: intensity of expression; patchiness of staining; presence of central artefact. Forty-six immunoreactions were inadequate (score 0 intensity), the majority regarding MLH1 or PMS2 in the hypo-fixation group (47.8%), followed by the hyper-fixation group (28.1%); cold formalin fixation showed the least inadequate cases. Patchiness and central artefact were more frequent in hypo-fixation and standard fixation group compared to the others. MLH1 (closely followed by PMS2) performed worse with regard to immunostaining intensity (p=0.0002) in the standard and in the hypo-fixation group (p< 0.00001). Using a small sample size improved patchiness/central artefacts. This is the first study specifically created to evaluate the impact of fixation on MMR protein IHC, showing that both formalin hypo- and hyper-fixation can cause problems; 24-h formalin fixation as well as cold (4°C) formalin fixation are recommended for successful IHC MMR evaluation. [ABSTRACT FROM AUTHOR]
Copyright of Virchows Archiv: European Journal of Pathology is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder's express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract. (Copyright applies to all Abstracts.)
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  Data: Impact of formalin fixation on mismatch repair protein evaluation by immunohistochemistry.
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  Data: Virchows Archiv: European Journal of Pathology; Nov2023, Vol. 483 Issue 5, p677-685, 9p
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  Label: Abstract
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  Data: Mismatch repair/microsatellite instability (MMR/MSI) status in colorectal cancer (CRC) has become fundamental as a diagnostic, prognostic, and predictive factor. MMR immunohistochemistry (IHC) is considered a simple and reliable approach; however, its effectiveness depends on pre-analytic factors. Aim of this study was to investigate the impact of different fixation times/protocols on MMR protein IHC quality. Left over tissue from surgically resected CRC samples (cold ischemia time &lt; 30 min) where fixed as follows: standard formalin fixation (24–48 h); hypo-fixation (&lt;20 h); hyper-fixation (&gt;90 h); cold (4&#176;C) fixation (24–48 h); standard fixation for small sample size (0.5&#215;0.5 cm). Samples for each group were collected from 30 resected CRC and the following parameters were evaluated on 600 immunohistochemical stains: intensity of expression; patchiness of staining; presence of central artefact. Forty-six immunoreactions were inadequate (score 0 intensity), the majority regarding MLH1 or PMS2 in the hypo-fixation group (47.8%), followed by the hyper-fixation group (28.1%); cold formalin fixation showed the least inadequate cases. Patchiness and central artefact were more frequent in hypo-fixation and standard fixation group compared to the others. MLH1 (closely followed by PMS2) performed worse with regard to immunostaining intensity (p=0.0002) in the standard and in the hypo-fixation group (p&lt; 0.00001). Using a small sample size improved patchiness/central artefacts. This is the first study specifically created to evaluate the impact of fixation on MMR protein IHC, showing that both formalin hypo- and hyper-fixation can cause problems; 24-h formalin fixation as well as cold (4&#176;C) formalin fixation are recommended for successful IHC MMR evaluation. [ABSTRACT FROM AUTHOR]
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  Label:
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  Data: &lt;i&gt;Copyright of Virchows Archiv: European Journal of Pathology is the property of Springer Nature and its content may not be copied or emailed to multiple sites or posted to a listserv without the copyright holder&#39;s express written permission. However, users may print, download, or email articles for individual use. This abstract may be abridged. No warranty is given about the accuracy of the copy. Users should refer to the original published version of the material for the full abstract.&lt;/i&gt; (Copyright applies to all Abstracts.)
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        Value: 10.1007/s00428-023-03661-z
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        Text: English
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            – D: 01
              M: 11
              Text: Nov2023
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