Combining Multiplex Fluorescence in situ Hybridization with Fluorescent Immunohistochemistry on Fresh Frozen or Fixed Mouse Brain Sections

10.3791/61709 ◽  
2021 ◽  
Author(s):  
Ayse S. Dereli ◽  
Evan J. Bailey ◽  
Natasha N. Kumar
Keyword(s):  
Mouse Brain ◽  
Fresh Frozen ◽  
Fluorescent Immunohistochemistry

A simplified method for combined immunohistochemistry and in-situ hybridization in fresh-frozen, cryocut mouse brain sections

2002 ◽  
Vol 9 (3) ◽  
pp. 214-219 ◽  
Author(s):  
Sathyanesan Samuel Newton ◽  
Antonia Dow ◽  
Rose Terwilliger ◽  
Ronald Duman
Keyword(s):  
In Situ Hybridization ◽  
Mouse Brain ◽  
Simplified Method ◽  
Fresh Frozen

scholarly journals Optimization and evaluation of fluorescence in situ hybridization chain reaction in cleared fresh-frozen brain tissues

2021 ◽  
Author(s):  
Vivek Kumar ◽  
David M. Krolewski ◽  
Elaine K. Hebda-Bauer ◽  
Aram Parsegian ◽  
Brian Martin ◽  
...  
Keyword(s):  
In Situ Hybridization ◽  
Fluorescence In Situ Hybridization ◽  
Hybridization Chain Reaction ◽  
Chain Reaction ◽  
Fresh Frozen ◽  
Brain Tissues

In situ hybridization study of the distribution of choline acetyltransferase mRNA and its splice variants in the mouse brain and spinal cord

Neuroscience ◽  
2009 ◽  
Vol 159 (1) ◽  
pp. 344-357 ◽  
Author(s):  
S. Trifonov ◽  
T. Houtani ◽  
S. Hamada ◽  
M. Kase ◽  
M. Maruyama ◽  
...  
Keyword(s):  
Spinal Cord ◽  
In Situ Hybridization ◽  
Mouse Brain ◽  
Choline Acetyltransferase ◽  
Splice Variants ◽  
Hybridization Study ◽  
Brain And Spinal Cord

scholarly journals Imaging Sterols and Oxysterols in Mouse Brain Reveals Distinct Spatial Cholesterol Metabolism

2018 ◽  
Author(s):  
Eylan Yutuc ◽  
Roberto Angelini ◽  
Mark Baumert ◽  
Natalia Mast ◽  
Irina Pikuleva ◽  
...  
Keyword(s):  
Mass Spectrometry ◽  
Mouse Brain ◽  
Brain Function ◽  
Mass Spectrometry Imaging ◽  
Cholesterol Metabolism ◽  
Biologically Active ◽  
Wild Type ◽  
Tissue Slices ◽  
The Brain

AbstractDysregulated cholesterol metabolism is implicated in a number of neurological disorders. Many sterols, including cholesterol and its precursors and metabolites, are biologically active and important for proper brain function. However, spatial cholesterol metabolism in brain and the resulting sterol distributions are poorly defined. To better understand cholesterol metabolism in situ across the complex functional regions of brain, we have developed on-tissue enzyme-assisted derivatisation in combination with micro-liquid-extraction for surface analysis and liquid chromatography - mass spectrometry to image sterols in tissue slices (10 µm) of mouse brain. The method provides sterolomic analysis at 400 µm spot diameter with a limit of quantification of 0.01 ng/mm2. It overcomes the limitations of previous mass spectrometry imaging techniques in analysis of low abundance and difficult to ionise sterol molecules, allowing isomer differentiation and structure identification. Here we demonstrate the spatial distribution and quantification of multiple sterols involved in cholesterol metabolic pathways in wild type and cholesterol 24S-hydroxylase knock-out mouse brain. The technology described provides a powerful tool for future studies of spatial cholesterol metabolism in healthy and diseased tissues.SignificanceThe brain is a remarkably complex organ and cholesterol homeostasis underpins brain function. It is known that cholesterol is not evenly distributed across different brain regions, however, the precise map of cholesterol metabolism in the brain remains unclear. If cholesterol metabolism is to be correlated with brain function it is essential to generate such a map. Here we describe an advanced mass spectrometry imaging platform to reveal spatial cholesterol metabolism in situ at 400 µm resolution on 10 µm tissue slices from mouse brain. We mapped, not only cholesterol, but also other biologically active sterols arising from cholesterol turnover in both wild type and mice lacking cholesterol 24-hydroxylase (Cyp46a1), the major cholesterol metabolising enzyme.

scholarly journals Evidence of Angiotensin Converting Enzyme 2 in Mouse Brain: in situ Hybridization and Mass Spectrometry Studies

2006 ◽  
Vol 20 (4) ◽  
Author(s):  
Yanfang Chen ◽  
Tatiana Cunha ◽  
Khalid Elased ◽  
Mariana Morris
Keyword(s):  
Mass Spectrometry ◽  
In Situ Hybridization ◽  
Angiotensin Converting Enzyme ◽  
Mouse Brain ◽  
Converting Enzyme ◽  
Angiotensin Converting Enzyme 2

scholarly journals Fluorescent In Situ Hybridization (FISH - RNAscope) in mouse brain sections v2 (protocols.io.btvenn3e)

protocols.io ◽  
2021 ◽  
Author(s):  
Yu Lin ◽  
Oriol Pavon ◽  
Lucille Duquenoy ◽  
Tiago Branco
Keyword(s):  
In Situ Hybridization ◽  
Mouse Brain ◽  
Fluorescent In Situ Hybridization

scholarly journals Fresh Frozen Mouse Brain Preparation (for Single Nuclei Sequencing) v1 (protocols.io.bcbrism6)

protocols.io ◽  
2020 ◽  
Author(s):  
Chuck Vanderburg ◽  
Carly Martin ◽  
Velina Kozareva ◽  
Naeem Nadaf ◽  
Nehal Patel ◽  
...  
Keyword(s):  
Mouse Brain ◽  
Fresh Frozen

Rapid β-oxidation of eicosapentaenoic acid in mouse brain: An in situ study

2009 ◽  
Vol 80 (2-3) ◽  
pp. 157-163 ◽  
Author(s):  
Chuck T. Chen ◽  
Zhen Liu ◽  
Melissa Ouellet ◽  
Frédéric Calon ◽  
Richard P. Bazinet
Keyword(s):  
Eicosapentaenoic Acid ◽  
Mouse Brain ◽  
In Situ Study

scholarly journals Comparing Biomechanical Properties, Repair Times, and Value of Common Core Flexor Tendon Repairs

Hand ◽  
2017 ◽  
Vol 13 (3) ◽  
pp. 313-318 ◽  
Author(s):  
Aakash Chauhan ◽  
Patrick Schimoler ◽  
Mark C. Miller ◽  
Alexander Kharlamov ◽  
Gregory A. Merrell ◽  
...  
Keyword(s):  
Flexor Tendon ◽  
Maximum Load ◽  
Biomechanical Properties ◽  
Barbed Suture ◽  
Load To Failure ◽  
Fresh Frozen ◽  
Post Hoc ◽  
Biomechanical Strength ◽  
Determining Factor

Background: The aim of the study was to compare biomechanical strength, repair times, and repair values for zone II core flexor tendon repairs. Methods: A total of 75 fresh-frozen human cadaveric flexor tendons were harvested from the index through small finger and randomized into one of 5 repair groups: 4-stranded cross-stitch cruciate (4-0 polyester and 4-0 braided suture), 4-stranded double Pennington (2-0 knotless barbed suture), 4-stranded Pennington (4-0 double-stranded braided suture), and 6-stranded modified Lim-Tsai (4-0 looped braided suture). Repairs were measured in situ and their repair times were measured. Tendons were linearly loaded to failure and multiple biomechanical values were measured. The repair value was calculated based on operating room costs, repair times, and suture costs. Analysis of variance (ANOVA) and Tukey post hoc statistical analysis were used to compare repair data. Results: The braided cruciate was the strongest repair ( P > .05) but the slowest ( P > .05), and the 4-stranded Pennington using double-stranded suture was the fastest ( P > .05) to perform. The total repair value was the highest for braided cruciate ( P > .05) compared with all other repairs. Barbed suture did not outperform any repairs in any categories. Conclusions: The braided cruciate was the strongest of the tested flexor tendon repairs. The 2-mm gapping and maximum load to failure for this repair approached similar historical strength of other 6- and 8-stranded repairs. In this study, suture cost was negligible in the overall repair cost and should be not a determining factor in choosing a repair.

The effect of in-situ decompression on ulnar nerve stability: a cadaveric study

2017 ◽  
Vol 42 (7) ◽  
pp. 715-719 ◽  
Author(s):  
B. Butler ◽  
J. Peelman ◽  
L.-Q. Zhang ◽  
M. Kwasny ◽  
D. Nagle
Keyword(s):  
Ulnar Nerve ◽  
Medial Epicondyle ◽  
Cadaveric Study ◽  
Flexion Angle ◽  
Anterior Translation ◽  
Nerve Decompression ◽  
Flexor Carpi Ulnaris ◽  
Before And After ◽  
Fresh Frozen

Ten fresh frozen right cadaver arms were placed in a motorized jig and an in-situ ulnar nerve decompression was performed in 5 mm increments distally to the flexor carpi ulnaris (FCU) aponeurosis then proximally to the intermuscular septum. The elbows were ranged 0–135° after each incremental decompression and the ulnar nerve to medial epicodyle distance was measured to assess for nerve translation/subluxation compared with baseline (prerelease) values. None of the specimens had ulnar nerve subluxation (defined as anterior translation past the medial epicondyle) even after full decompression. Furthermore, there were no statistically significant ulnar nerve translations (defined as any difference in distance from ulnar nerve to medial epicondyle before and after each decompression) for any flexion angle or extent of decompression. This study provides biomechanical evidence that in situ ulnar nerve decompression from the FCU aponeurosis to the intermuscular septum does not result in significant ulnar nerve translation or subluxation.

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