Ultra-rapid microwave-stimulated tissue processing with a modified protocol incorporating microwave fixation

Rapid tissue processing for transmission electron microscopy is a desired goal in a clinical EM laboratory. Reducing the time from tissue submission to examination in the electron microscope shortens turnaround time for diagnosis. Microwave enhanced tissue processing will accomplish these goals.Microwave technology reduces the time for tissue fixation and processing and may improve overall results. Fixation times can be reduced from minutes to seconds and resin polymerization from hours to minutes. Dehydration and infiltration steps can also be reduced, so that microwave rapid tissue processing can yield ultrathin sections from unfixed tissue in three hours.Three microwave protocols to process skeletal muscle biopsies are compared to standard processing. The protocols test the efficacy of microwave fixation, dehydration, and resin infiltration and polymerization. Protocol #1 uses microwave enhanced 4:1 formaldehyde/glutaraldehyde primary fixation followed by routine processing for remaining steps. Protocol #2 uses microwave primary fixation and 2% OsO4 post-fixation followed by routine processing. Protocol #3 uses microwave enhanced processing for entire procedure. Protocol #4 uses the standard processing technique.

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Opening images of synaptic vesicles and calcium localization by means of microwave fixation method

Proceedings, annual meeting, Electron Microscopy Society of America ◽

10.1017/s0424820100134508 ◽

1990 ◽

Vol 48 (2) ◽

pp. 182-183

Author(s):

Vinci Mizuhira ◽

Hiroshi Hasegawa

Keyword(s):

Synaptic Vesicles ◽

Room Temperature ◽

Osmium Tetroxide ◽

Sampling Procedure ◽

Fixation Method ◽

Potassium Oxalate ◽

X Ray ◽

Cacodylate Buffer ◽

Ultrathin Sections ◽

Microwave Fixation

Microwave irradiation (MWI) was applied to 0.3 to 1 cm3 blocks of rat central nervous system at 2.45 GHz/500W for about 20 sec in a fixative, at room temperature. Fixative composed of 2% paraformaldehyde, 0.5% glutaraldehyde in 0.1 M cacodylate buffer at pH 7.4, also contained 2 mM of CaCl2 , 1 mM of MgCl2, and 0.1% of tannic acid for conventional observation; and fuether 30-90 mM of potassium oxalate containing fixative was applied for the detection of calcium ion localization in cells. Tissue blocks were left in the same fixative for 30 to 180 min after MWI at room temperature, then proceeded to the sampling procedure, after postfixed with osmium tetroxide, embedded in Epon. Ultrathin sections were double stained with an useal manner. Oxalate treated sections were devided in two, stained and unstained one. The later oxalate treated unstained sections were analyzed with electron probe X-ray microanalyzer, the EDAX-PU-9800, at 40 KV accelerating voltage for 100 to 200 sec with point or selected area analyzing methods.

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Faculty Opinions recommendation of Microdissection testicular sperm extraction: preoperative patient optimization, surgical technique, and tissue processing.

Faculty Opinions – Post-Publication Peer Review of the Biomedical Literature ◽

10.3410/f.735241290.793564483 ◽

2019 ◽

Author(s):

Jay Sandlow

Keyword(s):

Surgical Technique ◽

Testicular Sperm Extraction ◽

Testicular Sperm ◽

Tissue Processing ◽

Patient Optimization ◽

Preoperative Patient

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Intramyocardial Transplantation of Autologous Myoblasts : Can Tissue Processing Be Optimized?

Circulation ◽

10.1161/01.cir.102.suppl_3.iii-210 ◽

2000 ◽

Vol 102 (Supplement 3) ◽

pp. III-210-III-215 ◽

Cited By ~ 46

Author(s):

B. Pouzet ◽

J.-T. Vilquin ◽

A. A. Hagege ◽

M. Scorsin ◽

E. Messas ◽

...

Keyword(s):

Tissue Processing ◽

Autologous Myoblasts

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A review of rapid microwave fixation technology: Its expanding niche in morphologic studies

Scanning ◽

10.1002/sca.4950150202 ◽

1993 ◽

Vol 15 (2) ◽

pp. 58-66 ◽

Cited By ~ 25

Author(s):

Gary R. Login ◽

Ann M. Dvorak

Keyword(s):

Microwave Fixation

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Letter to Editor: We Need Standardized Guidelines for Laboratory Tissue Processing After Microsurgical Testicular Sperm Extraction

Reproductive Sciences ◽

10.1007/s43032-021-00585-4 ◽

2021 ◽

Author(s):

Nader Salama

Keyword(s):

Testicular Sperm Extraction ◽

Testicular Sperm ◽

Tissue Processing

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Microfluidic platform accelerates tissue processing into single cells for molecular analysis and primary culture models

Nature Communications ◽

10.1038/s41467-021-23238-1 ◽

2021 ◽

Vol 12 (1) ◽

Author(s):

Jeremy A. Lombardo ◽

Marzieh Aliaghaei ◽

Quy H. Nguyen ◽

Kai Kessenbrock ◽

Jered B. Haun

Keyword(s):

Single Cell ◽

Single Cell Analysis ◽

Single Cells ◽

Cell Analysis ◽

Processing Technologies ◽

Microfluidic Platform ◽

Tissue Processing ◽

Single Cell Rna Sequencing ◽

Culture Models ◽

Tissue Digestion

AbstractTissues are complex mixtures of different cell subtypes, and this diversity is increasingly characterized using high-throughput single cell analysis methods. However, these efforts are hindered, as tissues must first be dissociated into single cell suspensions using methods that are often inefficient, labor-intensive, highly variable, and potentially biased towards certain cell subtypes. Here, we present a microfluidic platform consisting of three tissue processing technologies that combine tissue digestion, disaggregation, and filtration. The platform is evaluated using a diverse array of tissues. For kidney and mammary tumor, microfluidic processing produces 2.5-fold more single cells. Single cell RNA sequencing further reveals that endothelial cells, fibroblasts, and basal epithelium are enriched without affecting stress response. For liver and heart, processing time is dramatically reduced. We also demonstrate that recovery of cells from the system at periodic intervals during processing increases hepatocyte and cardiomyocyte numbers, as well as increases reproducibility from batch-to-batch for all tissues.

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