Freeze Substitution and Freeze Drying for Stable, Long-Term Preservation of Cytologic Specimens for Immunostaining

Takahashi, Rei; Okuyama, T.; Matsuo, Shingo; Maeda, Sakan

doi:10.1159/000333843

Cited by 3 publications

(1 citation statement)

References 14 publications

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“…In immunohistochemical analysis, antibodies raised against the c‐ erb B‐2, p53, and PCNA proteins reacted with each protein in the lyophilized tissue sample. These results are consistent with previous reports that p53 immunostaining is well retained in freeze‐dried cytological smear samples 20 , 21 . We anticipated that freezing in and of itself would not significantly hinder the formation of immune complexes in tissue samples.…”

Section: Discussionsupporting

confidence: 94%

Preservation of pathological tissue specimens by freeze‐drying for immunohistochemical staining and various molecular biological analyses

Matsuo

Sugiyama

Okuyama

et al. 1999

Pathology International

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Conditions of preserving DNA, RNA and protein in pathological specimens are of great importance as degradation of such macromolecules would critically affect results of molecular biological analysis. The feasibility of freeze-drying as a means of preserving pathological tissue samples for molecular analysis has previously been shown. In the present study, further tests on long-term storage conditions and analyses of freeze-dried samples by polymerase chain reaction (PCR), reverse transcriptase (RT)-PCR, western blotting and immunohistochemistry are reported. Rat chromosomal DNA of freeze-dried samples stored for 4 years showed slight degradation while RNA degradation was more prominently seen at an earlier stage of storage. However, these 4 year DNA and RNA samples were still able to serve as a template for some PCR and RT-PCR analyses, respectively. Overexpression of c-erbB-2 and p53 protein was demonstrated by western blotting and immunohistochemical staining using freeze-dried human breast cancer tissues. Although macromolecules in freeze-dried samples degrade to some extent during the preservation period, they should still be of value for certain molecular biological analyses and morphological examination; hence, providing more convenient and inexpensive ways of pathological tissue storage.

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Section: Discussionsupporting

confidence: 94%