A simplified and rapid procedure for in situ hybridization on human, flash-frozen, post-mortem brain and its combination with immunohistochemistry

1996 ◽  
Vol 69 (2) ◽  
pp. 213-227 ◽  
Author(s):  
Ann E. Kingsbury ◽  
E. Louise Bray ◽  
Oliver J.F. Foster
Keyword(s):  
In Situ Hybridization ◽  
Post Mortem ◽  
Rapid Procedure ◽  
Post Mortem Brain

A rapid procedure for the isolation of C0t-1 DNA from plants

Genome ◽  
1997 ◽  
Vol 40 (1) ◽  
pp. 138-142 ◽  
Author(s):  
Michael S. Zwick ◽  
Robert E. Hanson ◽  
M. Nurul Islam-Faridi ◽  
David M. Stelly ◽  
Rod A. Wing ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Repetitive Dna ◽  
Copy Number ◽  
Genomic Dna ◽  
Mammalian Species ◽  
Repetitive Dnas ◽  
Rapid Procedure ◽  
Dna Elements ◽  
Repetitive Dna Elements

In situ hybridization (ISH) for the detection of single- or low-copy sequences, particularly large DNA fragments cloned into YAC or BAC vectors, generally requires the suppression or "blocking" of highly-repetitive DNAs. C0t-1 DNA is enriched for repetitive DNA elements, high or moderate in copy number, and can therefore be used more effectively than total genomic DNA to prehybridize and competitively hybridize repetitive elements that would otherwise cause nonspecific hybridization. C0t-1 DNAs from several mammalian species are commercially available, however, none is currently available for plants to the best of our knowledge. We have developed a simple 1-day procedure to generate C0t-1 DNA without the use of specialized equipment.Key words: C0t-1 DNA, in situ hybridization, BACs, plants.

scholarly journals MO134COVID-19-ASSOCIATED KIDNEY INJURY IS CHARACTERIZED BY ACUTE TUBULAR NECROSIS AND CAPILLARY CONGESTION WITH EVIDENCE FOR SARS-COV-2 IN THE NEPHRON

2021 ◽  
Vol 36 (Supplement_1) ◽  
Author(s):  
Antoine Bouquegneau ◽  
Pauline Erpicum ◽  
Stéphanie Grosch ◽  
Lionel Habran ◽  
Olivier Hougrand ◽  
...  
Keyword(s):  
Electron Microscopy ◽  
Acute Kidney Injury ◽  
In Situ Hybridization ◽  
Acute Tubular Necrosis ◽  
Kidney Injury ◽  
Post Mortem ◽  
Rt Pcr ◽  
N Protein ◽  
Tubular Necrosis

Abstract Background and Aims Kidney damage has been reported in COVID-19 patients. Despite numerous reports about COVID-19-associated nephropathy, the factual presence of the SARS-CoV-2 in the renal parenchyma remains controversial. Method We consecutively performed 16 immediate (≤3h) post-mortem renal biopsies in patients diagnosed with COVID-19. Kidney samples from 5 patients who died from sepsis and were free from COVID-19 were used as controls. Samples were methodically evaluated by 3 pathologists. Virus detection in the renal parenchyma was performed in all samples by bulk RNA RT-PCR (E and N1/N2 genes), immunostaining (nCoV2019 N-Protein), fluorescent in situ hybridization (nCoV2019-S) and electron microscopy. Results The mean age of our COVID-19 cohort was 68.2±12.8 years, most of whom were males (68.7%). Proteinuria was observed in 53.3% of cases, while acute kidney injury occurred in 60% of cases. Acute tubular necrosis of variable severity was found in all cases, with no tubular or interstitial inflammation. There was no difference in acute tubular necrosis severity between the patients with COVID-19 versus control samples. Congestion in glomerular and peri-tubular capillaries was respectively observed in 56.3 and 87.5% of patients with COVID-19 compared to 20% of controls, with no evidence of thrombi. The nCoV2019 N-Protein was detected in proximal tubules and also at the basolateral pole of scattered cells of the distal tubules in 9/16 cases. In situ hybridization confirmed these findings. RT-PCR of kidney total RNA detected SARS-CoV-2 N gene in one case. Electron microscopy did not show typical viral inclusions. Conclusion Our immediate post-mortem kidney samples from patients with COVID-19 highlight a congestive pattern of acute kidney injury, with no significant glomerular or interstitial inflammation. Immunostaining and in situ hybridization suggest that SARS-CoV-2 is present in various segments of the nephron.

scholarly journals SARS-CoV-2 Infection in the Central Nervous System of a 1-Year-Old Infant Submitted to Complete Autopsy

2020 ◽  
Author(s):  
Ismael Carlos Gomes ◽  
Karina Karmirian ◽  
Julia Oliveira ◽  
Carolina Pedrosa ◽  
Fernando Colonna Rosman ◽  
...  
Keyword(s):  
Respiratory Illness ◽  
Organ Damage ◽  
Multiple Organ ◽  
Post Mortem ◽  
Cerebral Damage ◽  
Virus Particles ◽  
Brain Infection ◽  
Post Mortem Brain ◽  
The Central Nervous System

Coronavirus disease 2019 (COVID-19) was initially characterized as a respiratory illness. Neurological manifestations were reported mostly in severely affected patients. Routes for brain infection and the presence of virus particles in situ have not been well described, raising controversy about how the virus causes neurological symptoms. Here, we report the autopsy findings of a 1-year old infant with COVID-19. In addition to pneumonitis, meningitis and multiple organ damage related to thrombosis, a previous encephalopathy may have contributed to additional cerebral damage. SARS-CoV-2 infected the choroid plexus, ventricles, and cerebral cortex. This is the first evidence of SARS-CoV-2 detection in an infant post-mortem brain.

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

scholarly journals Post mortem brain temperature and its influence on quantitative MRI of the brain

2021 ◽  
Author(s):  
Celine Berger ◽  
Melanie Bauer ◽  
Holger Wittig ◽  
Eva Scheurer ◽  
Claudia Lenz
Keyword(s):  
Linear Models ◽  
Brain Temperature ◽  
Brain Mri ◽  
Mean Diffusivity ◽  
Quantitative Mri ◽  
Positive Temperature ◽  
Post Mortem ◽  
Post Mortem Brain

Abstract Objective MRI temperature sensitivity presents a major issue in in situ post mortem MRI (PMMRI), as the tissue temperatures differ from living persons due to passive cooling of the deceased. This study aims at computing brain temperature effects on the MRI parameters to correct for temperature in PMMRI, laying the foundation for future projects on post mortem validation of in vivo MRI techniques. Materials and methods Brain MRI parameters were assessed in vivo and in situ post mortem using a 3 T MRI scanner. Post mortem brain temperature was measured in situ transethmoidally. The temperature effect was computed by fitting a linear model to the MRI parameters and the corresponding brain temperature. Results Linear positive temperature correlations were observed for T1, T2* and mean diffusivity in all tissue types. A significant negative correlation was observed for T2 in white matter. Fractional anisotropy revealed significant correlations in all gray matter regions except for the thalamus. Discussion The linear models will allow to correct for temperature in post mortem MRI. Comparing in vivo to post mortem conditions, the mean diffusivity, in contrast to T1 and T2, revealed additional effects besides temperature, such as cessation of perfusion and active diffusion.

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