scholarly journals Cryosectioning, block face imaging and Nissl staining fluorescently labeled pig heart

2021 ◽  
Author(s):  
Shaina Robbins ◽  
Rajanikanth Vadigepalli ◽  
James Schwaber
Keyword(s):  
Molecular Data ◽  
Heart Tissue ◽  
Single Neurons ◽  
Nissl Staining ◽  
Microscopic Imaging ◽  
Tissue Sections ◽  
Face Images ◽  
Fresh Frozen ◽  
Block Face ◽  
Laser Capture

Abstract The present protocol describes the process of cryosectioning OCT-embedded fresh frozen pig heart tissue, along with the acquisition of block face images, followed by Nissl staining of the tissue sections and microscopic imaging of the sections for localizing the fluorescently-labeled neurons in the tissue. This tissue section preparation workflow was optimized for use in laser capture microdissection of single neurons from the pig heart tissue, which are used in downstream transcriptomic analysis and mapping of the neuronal location and molecular data to a 3D reconstructed contour stack of block face images.

scholarly journals Single-Cell Mapping and 3D Tissue Reconstruction using Cryosection-derived Images and Tissue Mapper software

2021 ◽  
Author(s):  
Shanina Robbins ◽  
Rajanikanth Vadigepalli ◽  
James Schwaber
Keyword(s):  
3D Visualization ◽  
Molecular Data ◽  
Single Neurons ◽  
Cell Mapping ◽  
Face Images ◽  
Tissue Reconstruction ◽  
Block Face ◽  
The Individual ◽  
Anatomical Information ◽  
Laser Capture

Abstract The present protocol describes the process of using the MBF Bioscience Tissue Mapper software to create a 3D reconstructed stack from contouring the block face images of serial cryosections and mark the individual immunolabeled cells onto the contoured images. The stack of aligned, contoured and annotated images can be visualized as a 3D reconstruction that depicts the tissue shape as it was embedded and cryosectioned, relative positions of various tissue features, and locations of individually marked neurons within the tissue, yielding a 3D anatomical map of the distribution of cells of interest with a tissue. This protocol can be used in conjunction with other protocols for sampling spatially-tracked single neurons using laser capture micro-dissection and assessed for molecular profiles using transcriptomics approaches. Such integration extends the present protocol to enable 3D visualization of molecular data in conjunction with the anatomical information for spatial transcriptomic analysis.

Detection of Cancer-Related Proteins in Fresh-Frozen Ovarian Cancer Samples Using Laser Capture Microdissection

2008 ◽  
pp. 35-45
Author(s):  
Dan-Arin Silasi ◽  
Ayesha B. Alvero ◽  
Jechiel Mor ◽  
Rui Chen ◽  
Han-Hsuan Fu ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Laser Capture Microdissection ◽  
Fresh Frozen ◽  
Related Proteins ◽  
Laser Capture

scholarly journals Fast and Simple Protocols for Mass Spectrometry-Based Proteomics of Small Fresh Frozen Uterine Tissue Sections

2017 ◽  
Vol 89 (20) ◽  
pp. 10769-10775 ◽  
Author(s):  
Irena Dapic ◽  
Naomi Uwugiaren ◽  
Petra J. Jansen ◽  
Garry L. Corthals
Keyword(s):  
Mass Spectrometry ◽  
Uterine Tissue ◽  
Tissue Sections ◽  
Fresh Frozen

Detection of Cancer-Related Proteins in Fresh-Frozen Ovarian Cancer Samples Using Laser Capture Microdissection

2008 ◽  
pp. 35-46
Author(s):  
Dan-Arin Silasi ◽  
Ayesha B. Alvero ◽  
Jechiel Mor ◽  
Rui Chen ◽  
Han-Hsuan Fu ◽  
...  
Keyword(s):  
Ovarian Cancer ◽  
Laser Capture Microdissection ◽  
Fresh Frozen ◽  
Related Proteins ◽  
Laser Capture

A bioartificial rat heart tissue: Perfusion decellularization and characterization

2019 ◽  
Vol 42 (12) ◽  
pp. 757-764 ◽  
Author(s):  
Busra Ozlu ◽  
Mert Ergin ◽  
Sevcan Budak ◽  
Selcuk Tunali ◽  
Nuh Yildirim ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Three Dimensional ◽  
Tissue Perfusion ◽  
Heart Tissue ◽  
Cell Nuclei ◽  
Tissue Sections ◽  
The Past ◽  
Significant Difference ◽  
Structural Preservation

Despite remarkable advancement in the past decades, heart-related defects are still prone to progress irreversibly and can eventually lead to heart failure. A personalized extracellular matrix–based bioartificial heart created by allografts/xenografts emerges as an alternative as it can retain the original three-dimensional architecture combined with a preserved natural heart extracellular matrix. This study aimed at developing a procedure for decellularizing heart tissue harvested from rats and evaluating decellularization efficiency in terms of residual nuclear content and structural properties. Tissue sections showed no or little visible cell nuclei in decellularized heart, whereas the native heart showed dense cellularity. In addition, there was no significant variation in the alignment of muscle fibers upon decellularization. Furthermore, no significant difference was detected between native and decellularized hearts in terms of fiber diameter. Our findings demonstrate that fiber alignment and diameter can serve as additional parameters in the characterization of biological heart scaffolds as these provide valuable input for evaluating structural preservation of decellularized heart. The bioartificial scaffold formed here can be functionalized with patient’s own material and utilized in regenerative engineering.

scholarly journals Brief Report: Simplified Myeloperoxidase Stain Using Benzidine Dihydrochloride

Blood ◽  
1965 ◽  
Vol 26 (2) ◽  
pp. 215-219 ◽  
Author(s):  
LEONARD S. KAPLOW ◽  
Charlotte Ladd
Keyword(s):  
Peroxidase Activity ◽  
Incubation Mixture ◽  
Tissue Sections ◽  
Blood Smears ◽  
Highly Sensitive ◽  
Frozen Tissue ◽  
Fresh Frozen

Abstract A method is described for demonstrating leukocyte peroxidase activity in which benzidine dihydrochloride is used as the indicator compound instead of the more commonly used but potentially more hazardous benzidine base. The method is highly sensitive and rapid and permits the use of fixed blood smears and organ imprints. The incubation mixture, which incorporates safranin as a counterstain, may be used over and over again, for as long as 6 months. The method is also applicable to fresh frozen tissue sections.

scholarly journals Tissue preparation for laser capture microdissection and RNA extraction from fresh frozen breast tissue

BioTechniques ◽  
2007 ◽  
Vol 43 (1) ◽  
pp. 41-48 ◽  
Author(s):  
Mary Morrogh ◽  
Narciso Olvera ◽  
Faina Bogomolniy ◽  
Patrick I. Borgen ◽  
Tari A. King
Keyword(s):  
Laser Capture Microdissection ◽  
Breast Tissue ◽  
Rna Extraction ◽  
Tissue Preparation ◽  
Fresh Frozen ◽  
Laser Capture
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