Post-mortem analysis of bone marrow osteoclasts using tartrate-resistant acid phosphatase staining: does histochemistry work and correlate with time since death?

2012 ◽  
Vol 65 (11) ◽  
pp. 1013-1018 ◽  
Author(s):  
Joachim Boehm ◽  
Ulrike Schmidt ◽  
Michéle Porsche ◽  
Juergen Veeck ◽  
Hans-Eckart Schaefer
Keyword(s):  
Bone Marrow ◽  
Acid Phosphatase ◽  
Staining Intensity ◽  
Tartrate Resistant Acid Phosphatase ◽  
Time Of Death ◽  
Post Mortem ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Trap Staining ◽  
Formalin Fixed

AimsIn bone marrow (BM) biopsies, tartrate-resistant acid phosphatase (TRAP) staining represents the gold standard for the characterisation of osteoclasts. TRAP is one of the few enzymes that is histochemically detectable on formalin-fixed paraffin-embedded tissue. This study investigated whether TRAP is also able to visualise BM osteoclasts in autopsy tissue. It was hypothesised that, due to a progressive loss of enzymatic activity in osteoclasts post-mortem, TRAP staining could allow the time of death of a patient to be determined.MethodsTRAP-stained BM slides of 96 cases including 51 pathology and 23 forensic autopsies and 22 biopsies were histologically evaluated and their staining intensity (SI) semi-quantitatively graded. In the autopsy cases, the results were correlated with the post-mortem interval (PMI, time span in days between death and autopsy).ResultsTRAP staining intensities (TRAP-SIs) did not differ between men and women and showed a steady decrease with age. TRAP-SIs were significantly stronger in biopsies than in autopsy cases. Among the autopsies, TRAP-SIs were highly variable and not dependent on PMI, except for three forensic cases with PMI ≥7 days which showed a complete loss of TRAP stainability. On the whole, the TRAP-SIs of pathology and forensic cases did not differ significantly.ConclusionsThis study clearly shows that BM osteoclasts stay TRAP-positive for 7 days post-mortem, although with markedly reduced TRAP-SIs compared with biopsies. Since TRAP-SIs were not correlated with the duration of PMI, TRAP staining of BM osteoclasts cannot serve as a tool to determine the time of death of a patient.

scholarly journals BRAFV600E-Positive Precursors As Molecular Markers of Bone Marrow Involvement in Pediatric Langerhans Cell Histiocytosis

Blood ◽  
2019 ◽  
Vol 134 (Supplement_1) ◽  
pp. 3588-3588
Author(s):  
Ko Kudo ◽  
Rika Kanezaki ◽  
Akie Kobayashi ◽  
Tomohiko Sato ◽  
Takuya Kamio ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Somatic Mutations ◽  
Bone Marrow Involvement ◽  
Braf V600e ◽  
Sensitive Assay ◽  
System Disease ◽  
Multiple Organs ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

Introduction: The BRAF mutation V600E, the most common somatic mutation in Langerhans cell histiocytosis (LCH), has been reported in approximately 50% of LCH patients and is associated with certain high-risk clinical features. Precursors harboring this mutation can differentiate into Langerhans cells resulting in infiltrates in multiple organs under inflammatory conditions. However, BRAF status in the bone marrow of pediatric LCH patients is unclear. The present study examined somatic mutations in paired tumor and bone marrow samples, using a highly sensitive assay involving next-generation targeted sequencing and droplet digital polymerase chain reaction (PCR) for pediatric LCH patients. Methods: Between 1996 and 2019, in total of 17 Japanese pediatric patients with LCH were enrolled. The male/female ratio was 7/11. Ages of onset of LCH were median 13 months (range 5-193 months). At diagnosis of LCH, 2 patients were positive for risk organ involvement, 15 were negative. We retrospectively performed mutational analyses of 17 LCH cases using formalin-fixed paraffin-embedded LCH tumor specimens to provide templates for PCR-based targeted amplicon sequencing with customized primers to detect mutations in exons 12 and 15 in BRAF, and exons 2 and 3 in MAP2K1. Thereafter, we identified somatic mutations in the 17 paired bone marrow samples via droplet digital allele-specific PCR, targeting BRAF V600E and BRAF exon 12 in-frame deletion 496-500 (Ex12 in-del). Results: We detected BRAF V600E in 11 of 17 tumor samples (65%) and the BRAF Ex 12 in-del in 3 of 17 tumors (18%). We identified BRAF V600E in bone marrow samples in 10 of the 11 cases (90%) with the mutation in the tumor at low variant allele frequency (median 0.25%, range 0.14-7.0%). BRAF Ex 12 in-del was not detected in the bone marrow. Cases with detectable bone marrow involvement included eight patients with multi-system disease affecting multiple organs, one patient with multi-focal bone disease, and one patient with single-system disease. Clinical phenotypes including relapse did not correlate with BRAF V600E upon detection in the bone marrow. Conclusion: We established the sensitive assay based on PCR-based targeted NGS for detecting somatic mutations in LCH even accessible for formalin-fixed, paraffin-embedded clinical specimens. Bone marrow involvement is frequently detectable at the molecular level in pediatric LCH with the BRAF V600E mutation. A prospective study is warranted to evaluate the clinical impact of mutational burden in bone marrow. Disclosures Kudo: Unum Therapeutics: Patents & Royalties. Imai:Juno Therapeutics: Patents & Royalties.

scholarly journals SARS-CoV-2 genome sequencing from post-mortem formalin-fixed, paraffin-embedded lung tissues

2021 ◽  
Author(s):  
Claude Van Campenhout ◽  
Ricardo De Mendonça ◽  
Barbara Alexiou ◽  
Sarah De Clercq ◽  
Marie-Lucie Racu ◽  
...  
Keyword(s):  
Genome Sequencing ◽  
Post Mortem ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

A quantitative in situ hybridization protocol for formalin-fixed paraffin-embedded archival post-mortem human brain tissue

Methods ◽  
2010 ◽  
Vol 52 (4) ◽  
pp. 359-366 ◽  
Author(s):  
Chun-Qing Liu ◽  
Ling Shan ◽  
Rawien Balesar ◽  
Sabina Luchetti ◽  
Joop J. Van Heerikhuize ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Human Brain ◽  
Brain Tissue ◽  
Post Mortem ◽  
Human Brain Tissue ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Hybridization Protocol ◽  
Formalin Fixed

The Post-Mortem Detection of Early Myocardial Infarction by a Simple Fluorescent Method

1976 ◽  
Vol 16 (1) ◽  
pp. 17-20 ◽  
Author(s):  
V. B. Sahai ◽  
Rnard Knight
Keyword(s):  
Myocardial Infarction ◽  
Hypertensive Heart Disease ◽  
Secondary Emission ◽  
Post Mortem ◽  
Simple Method ◽  
Fluorescent Method ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Artery Disease ◽  
Formalin Fixed

A rapid and simple method is described for demonstrating early ischaemic/anoxic damage to the myocardium due either to coronary artery disease or hypertensive heart disease. Using formalin-fixed paraffin-embedded standard histological sections, staining for only 3 seconds in an aqueous solution of acridine-orange was sufficient to demonstrate a green shift of the secondary emission produced when the section was viewed in ultraviolet light. The results correlated well with sensitive enzyme studies which, however, were technically more difficult. The ease of retaining tissues and the rapidity of the technique, coupled with the reliability of the method even in the presence of moderate post-mortem autolysis, makes this a useful method for routine use.

scholarly journals Mass Spectrometry Amyloid Typing Is Reproducible across Multiple Organ Sites

2019 ◽  
Vol 2019 ◽  
pp. 1-9 ◽  
Author(s):  
Dusan Holub ◽  
Pavla Flodrova ◽  
Tomas Pika ◽  
Patrik Flodr ◽  
Marian Hajduch ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Staining Intensity ◽  
Multiple Organ ◽  
Ffpe Tissue ◽  
Al Amyloidosis ◽  
Tissue Samples ◽  
Amyloid A ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Formalin Fixed

We have determined patient’s amyloid subtype through immunohistochemical and proteomic analyses of formalin-fixed, paraffin-embedded (FFPE) tissue samples from two affected organs per patient. Amyloid typing, via immunohistochemistry (IHC) and laser microdissection followed by the combination of liquid chromatography with mass spectrometry (LMD-LC-MS), was performed using tissue samples of the human heart, liver, kidney, tongue, and small intestine from 11 patients, and the results were compared with clinical data. LMD-LC-MS correctly typed AL amyloidosis in all 22 FFPE tissue samples despite tissue origin. In contrast, IHC was successful only in the analysis of eight FFPE tissue samples with differences between the examined organs. In the majority of LMD-LC-MS typed samples, the level of IHC staining intensity for transthyretin and serum amyloid A was the same as that for Ig κ and Ig λ antibodies, suggesting low Ig κ or Ig λ antibodies reactivity and the additional antibody clones were essential for correct typing. Both methods used in the study were found to be suitable for amyloid typing, although LMD-LC-MS yielded more promising results than IHC.

Impact of Preanalytical Factors on the Measurement of Tumor Tissue Biomarkers Using Immunohistochemistry

2021 ◽  
pp. 002215542199560
Author(s):  
Aditi Bagchi ◽  
Zachary Madaj ◽  
Kelly B. Engel ◽  
Ping Guan ◽  
Daniel C. Rohrer ◽  
...  
Keyword(s):  
Tumor Tissue ◽  
Cancer Biomarkers ◽  
Staining Intensity ◽  
Ffpe Tissue ◽  
Kidney Tumor ◽  
Formalin Fixed Paraffin ◽  
Formalin Fixed Paraffin Embedded ◽  
Tumor Types ◽  
Formalin Fixed ◽  
Tissue Biomarkers

Analysis of formalin-fixed paraffin-embedded (FFPE) tissue by immunohistochemistry (IHC) is commonplace in clinical and research laboratories. However, reports suggest that IHC results can be compromised by biospecimen preanalytical factors. The National Cancer Institute’s Biospecimen Preanalytical Variables Program conducted a systematic study to examine the potential effects of delay to fixation (DTF) and time in fixative (TIF) on IHC using 24 cancer biomarkers. Differences in IHC staining, relative to controls with a DTF of 1 hr, were observed in FFPE kidney tumor specimens after a DTF of ≥2 hr. Reductions in H-score and/or staining intensity were observed for c-MET, p53, PAX2, PAX8, pAKT, and survivin, whereas increases were observed for RCC1, EGFR, and CD10. Prolonged TIF of 72 hr resulted in significantly reduced H-scores of CD44 and c-Met in kidney tumor specimens, compared with controls with 12-hr TIF. An elevated probability of altered staining intensity due to DTF was observed for nine antigens, whereas for prolonged TIF an elevated probability was observed for one antigen. Results reported here and elsewhere across tumor types and antigens support limiting DTF to ≤1 hr when possible and fixing tissues in formalin for 12–24 hr to avoid confounding effects of these preanalytical factors on IHC.

Sign in / Sign up

Export Citation Format

Share Document