Next Article in Journal
Emerging Endorobotic and AI Technologies in Colorectal Cancer Screening: A Review of Design, Validation, and Translational Pathways
Next Article in Special Issue
Fatal Choking in Patients with Psychiatric Disorders: A Systematic Review of Forensic Evidence
Previous Article in Journal
BanglaOCT2025: A Population-Specific Fovea-Centric OCT Dataset with Self-Supervised Volumetric Restoration Using Flip-Flop Swin Transformers
Previous Article in Special Issue
Postmortem Microbiology in Forensic Diagnostics: Interpretation of Infectious Causes of Death and Emerging Applications
 
 
Font Type:
Arial Georgia Verdana
Font Size:
Aa Aa Aa
Line Spacing:
Column Width:
Background:
Article

The Shrinking Blind Spot: How Freeze–Thaw Obscures Microscopic Evidence of Ante-Mortem Ecchymosis

Department of Surgical Pathology, Medical, Molecular, and Critical Area, Institute of Legal Medicine, University of Pisa, 56126 Pisa, Italy
*
Author to whom correspondence should be addressed.
Diagnostics 2026, 16(3), 419; https://doi.org/10.3390/diagnostics16030419
Submission received: 23 December 2025 / Revised: 19 January 2026 / Accepted: 21 January 2026 / Published: 1 February 2026
(This article belongs to the Special Issue Diagnostic Methods in Forensic Pathology, Third Edition)

Abstract

Background/Objectives: Histological examination constitutes a fundamental methodology for establishing the vitality of a lesion. In cases where the corpse is preserved for an extended duration of time prior to the post-mortem evaluation, particularly if the body has undergone freezing and thawing cycles, post-mortem changes may obscure or alter evidence of traumatic injuries. Consequently, the reliability of hematoxylin and eosin (H&E) staining for the reliable detection of intralesional erythrocytes in suspected traumatic fatalities is potentially severely compromised. The primary objective of this study is to rigorously underscore the detrimental influence of freeze–thaw processes on histologic examination and to advocate the indispensable incorporation of immunohistochemical analysis, specifically employing anti-human glycophorin A antibodies, to ascertain the presence of red blood cells. Methods: Skin samples from 10 autopsy cases were subjected to serial freeze–thaw cycles and analyzed using anti-human Glycophorin A (GPA) immunohistochemistry staining to evaluate skin lesion vitality in freeze–thawed tissues compared to fresh controls. Results: Results indicated that while H&E reliability was limited to fresh tissue, anti-GPA staining remained stable across all freeze–thaw cycles. Conclusions: Forensic pathologists must remain acutely cognizant of the potential artifacts produced by freeze–thaw cycles. In these cases, anti-GPA staining proved to be a reliable asset for evaluating the vitality of a lesion.
Keywords: glycophorin; immunohistochemistry; red blood cells; freeze–thaw cycles; autopsy artifacts glycophorin; immunohistochemistry; red blood cells; freeze–thaw cycles; autopsy artifacts

Share and Cite

MDPI and ACS Style

Iacoponi, N.; Giacomelli, S.; Turillazzi, E.; Di Paolo, M. The Shrinking Blind Spot: How Freeze–Thaw Obscures Microscopic Evidence of Ante-Mortem Ecchymosis. Diagnostics 2026, 16, 419. https://doi.org/10.3390/diagnostics16030419

AMA Style

Iacoponi N, Giacomelli S, Turillazzi E, Di Paolo M. The Shrinking Blind Spot: How Freeze–Thaw Obscures Microscopic Evidence of Ante-Mortem Ecchymosis. Diagnostics. 2026; 16(3):419. https://doi.org/10.3390/diagnostics16030419

Chicago/Turabian Style

Iacoponi, Naomi, Sara Giacomelli, Emanuela Turillazzi, and Marco Di Paolo. 2026. "The Shrinking Blind Spot: How Freeze–Thaw Obscures Microscopic Evidence of Ante-Mortem Ecchymosis" Diagnostics 16, no. 3: 419. https://doi.org/10.3390/diagnostics16030419

APA Style

Iacoponi, N., Giacomelli, S., Turillazzi, E., & Di Paolo, M. (2026). The Shrinking Blind Spot: How Freeze–Thaw Obscures Microscopic Evidence of Ante-Mortem Ecchymosis. Diagnostics, 16(3), 419. https://doi.org/10.3390/diagnostics16030419

Note that from the first issue of 2016, this journal uses article numbers instead of page numbers. See further details here.

Article Metrics

Back to TopTop