Capillary electrophoretic measurement of tissue metabolites

1998 ◽  
Vol 274 (3) ◽  
pp. C840-C845 ◽  
Author(s):  
Patrick F. Dillon ◽  
Patrick R. Sears
Keyword(s):  
Enzymatic Analysis ◽  
Similar Time ◽  
Tissue Samples ◽  
Phosphorus Nmr ◽  
Rapid Measurement ◽  
Capillary Electrophoretic ◽  
Multiple Samples ◽  
Rabbit Urinary Bladder ◽  
Electrophoretic Measurement ◽  
Tissue Metabolites

A method for the measurement of tissue metabolites from rabbit urinary bladder using capillary electrophoresis (CE) has been developed. The method generates a reproducible electropherogram containing >20 peaks, including NAD, NADH, lactate, UDP-glucose, phosphocreatine, creatine, ATP, ADP, GTP, and UTP, from <20 nl of extract solution generated from 1.1 nl (or ∼1.2 μg) of tissue in <40 min. Multiple samples from the same bladder produce SE comparable with enzymatic or nuclear magnetic resonance (NMR) measurements of metabolites: phosphorus-NMR measurement requires 106 more tissue than CE; individual enzymatic measurements using 100 μl/sample require 2,000 μl, a 105 greater volume than required by CE for the same number of metabolites. CE detects about three times more peaks than phosphorus-NMR on a similar time scale. Comparable measurements using enzymatic analysis would require ∼10 times longer. The combination of minimal tissue volume requirements, rapid measurement, and reproducibility makes CE a valuable tool in the investigation of simultaneous changes in multiple metabolites from minute tissue samples.

scholarly journals High-Throughput Strategy for Profiling Sequential Section With Multiplex Staining of Mouse Brain

2021 ◽  
Vol 15 ◽  
Author(s):  
Siqi Chen ◽  
Zhixiang Liu ◽  
Anan Li ◽  
Hui Gong ◽  
Ben Long ◽  
...  
Keyword(s):  
Mouse Brain ◽  
Acute Stress ◽  
Brain Slices ◽  
Serial Sections ◽  
Whole Brain ◽  
Tissue Samples ◽  
Brain Functions ◽  
Multiple Samples ◽  
The Brain ◽  
Serial Brain

The brain modulates specific functions in its various regions. Understanding the organization of different cells in the whole brain is crucial for investigating brain functions. Previous studies have focused on several regions and have had difficulty analyzing serial tissue samples. In this study, we introduced a pipeline to acquire anatomical and histological information quickly and efficiently from serial sections. First, we developed a serial brain-slice-staining method to stain serial sections and obtained more than 98.5% of slices with high integrity. Subsequently, using the self-developed analysis software, we registered and quantified the signals of imaged sections to the Allen Mouse Brain Common Coordinate Framework, which is compatible with multimodal images and slant section planes. Finally, we validated the pipeline with immunostaining by analyzing the activity variance in the whole brain during acute stress in aging and young mice. By removing the problems resulting from repeated manual operations, this pipeline is widely applicable to serial brain slices from multiple samples in a rapid and convenient manner, which benefits to facilitate research in life sciences.

scholarly journals Is it necessary to change instruments between sampling sites when taking multiple tissue specimens in musculoskeletal infections?

2018 ◽  
Vol 100 (7) ◽  
pp. 563-565 ◽  
Author(s):  
D Makki ◽  
S Abdalla ◽  
TA El Gamal ◽  
D Harvey ◽  
G Jackson ◽  
...  
Keyword(s):  
Sampling Site ◽  
Surgical Debridement ◽  
Cross Contamination ◽  
Tissue Samples ◽  
Musculoskeletal Infection ◽  
Exact Test ◽  
Biopsy Site ◽  
Orthopaedic Infections ◽  
Tissue Specimens ◽  
Multiple Samples

Introduction Surgical debridement of orthopaedic infections allows biopsy for microbiology and facilitates successful treatment. It is recommended that biopsy instruments are changed when taking multiple samples. This study compared assessed cross-contamination between biopsy sites when using same instruments to take tissue samples from multiple sites. Materials and methods During the surgical debridement, we defined five sampling sites and marked them with diathermy. Two sampling techniques were performed on same patient to minimise any potential bias arising from the type of host and the severity of infection. First, fresh instruments were used for each biopsy site. Titleond, the instruments used in the first sampling site were reused to take samples from the remaining sites. By comparing the microbiology results of the samples taken by each technique for each site we determined cross-contamination with microorganisms. Results Fifteen patients with foot and ankle infections (mean age 56 years) were included. Ten patients were diabetic and five had neuropathies. Cross-contamination between sampling sites occurred in eight cases when the same instruments were used to take biopsies (P = 0.002, Fisher’s exact test). One or more microorganisms were involved in cross-contamination and the latter always occurred between two consecutive sites rather than sites that were further apart. Conclusion It is important to use fresh instruments for each biopsy site when taking multiple samples in musculoskeletal infection as cross-contamination might occur otherwise and affect microbiological studies.

scholarly journals Digital Staining of High-Definition Fourier Transform Infrared (FT-IR) Images Using Deep Learning

2019 ◽  
Vol 73 (5) ◽  
pp. 556-564 ◽  
Author(s):  
Mahsa Lotfollahi ◽  
Sebastian Berisha ◽  
Davar Daeinejad ◽  
David Mayerich
Keyword(s):  
Fourier Transform ◽  
Fourier Transform Infrared ◽  
Cellular Level ◽  
Label Free ◽  
High Definition ◽  
Tissue Samples ◽  
Ft Ir ◽  
Downstream Analysis ◽  
Ir Spectroscopic ◽  
Multiple Samples

Histological stains, such as hematoxylin and eosin (H&E), are routinely used in clinical diagnosis and research. While these labels offer a high degree of specificity, throughput is limited by the need for multiple samples. Traditional histology stains, such as immunohistochemical labels, also rely only on protein expression and cannot quantify small molecules and metabolites that may aid in diagnosis. Finally, chemical stains and dyes permanently alter the tissue, making downstream analysis impossible. Fourier transform infrared (FT-IR) spectroscopic imaging has shown promise for label-free characterization of important tissue phenotypes and can bypass the need for many chemical labels. Fourier transform infrared classification commonly leverages supervised learning, requiring human annotation that is tedious and prone to errors. One alternative is digital staining, which leverages machine learning to map IR spectra to a corresponding chemical stain. This replaces human annotation with computer-aided alignment. Previous work relies on alignment of adjacent serial tissue sections. Since the tissue samples are not identical at the cellular level, this technique cannot be applied to high-definition FT-IR images. In this paper, we demonstrate that cellular-level mapping can be accomplished using identical samples for both FT-IR and chemical labels. In addition, higher-resolution results can be achieved using a deep convolutional neural network that integrates spatial and spectral features.

scholarly journals A Modified Fluorimetric Method for Determination of Hydrogen Peroxide Using Homovanillic Acid Oxidation Principle

2014 ◽  
Vol 2014 ◽  
pp. 1-8 ◽  
Author(s):  
Biswaranjan Paital
Keyword(s):  
Hydrogen Peroxide ◽  
Biological Samples ◽  
Homovanillic Acid ◽  
Low Cost ◽  
Internal Standard ◽  
Acid Oxidation ◽  
Tissue Samples ◽  
The Stability ◽  
Multiple Samples

Hydrogen peroxide (H2O2) level in biological samples is used as an important index in various studies. Quantification of H2O2level in tissue fractions in presence of H2O2metabolizing enzymes may always provide an incorrect result. A modification is proposed for the spectrofluorimetric determination of H2O2in homovanillic acid (HVA) oxidation method. The modification was included to precipitate biological samples with cold trichloroacetic acid (TCA, 5% w/v) followed by its neutralization with K2HPO4before the fluorimetric estimation of H2O2is performed. TCA was used to precipitate the protein portions contained in the tissue fractions. After employing the above modification, it was observed that H2O2content in tissue samples was ≥2 fold higher than the content observed in unmodified method. Minimum 2 h incubation of samples in reaction mixture was required for completion of the reaction. The stability of the HVA dimer as reaction product was found to be >12 h. The method was validated by using known concentrations of H2O2and catalase enzyme that quenches H2O2as substrate. This method can be used efficiently to determine more accurate tissue H2O2level without using internal standard and multiple samples can be processed at a time with additional low cost reagents such as TCA and K2HPO4.

Manual Determination of Intra-Erythrocytic 2,3-Diphosphoglycerate in Whole Blood by Enzymatic Analysis, and Comparison with Ion-Exchange Chromatography

1975 ◽  
Vol 21 (3) ◽  
pp. 376-380 ◽  
Author(s):  
James C Detter ◽  
Donald F Gibson ◽  
Stuart F MacMillan ◽  
Thomas H Oas
Keyword(s):  
Ion Exchange ◽  
Colorimetric Assay ◽  
Ion Exchange Chromatography ◽  
Exchange Chromatography ◽  
Single Subject ◽  
Enzymatic Analysis ◽  
Automated Method ◽  
Enzyme Method ◽  
Multiple Samples

Abstract A manual modification of an automated method [Atkinson, K. F., Clin. Chem. 18, 1001 (1972)] for enzymatic assay of 2,3-diphosphoglycerate is described, which is suitable for small laboratories. Samples are easily prepared for analysis, and preparations are stable for several months. Virtually all of the color generated in the colorimetric assay is produced by the diphosphoglycerate-catalyzed reaction. The coefficient of variation for multiple samples from a single subject was 2.4%. Delayed preparation of samples, particularly samples from some acidotic subjects, is shown to result in altered results. The enzyme method is compared with analysis by ion-exchange chromatography.

In situ microprobe quantitation of inorganic particle burden in paraffin sections of lung tissue

1988 ◽  
Vol 46 ◽  
pp. 990-991
Author(s):  
Jerrold L. Abraham
Keyword(s):  
Lung Tissue ◽  
Quantitative Information ◽  
Particulate Material ◽  
Paraffin Sections ◽  
Tissue Samples ◽  
Qualitative And Quantitative ◽  
The Past ◽  
Quantitative Analyses ◽  
Data Result

Inorganic particulate material of diverse types is present in the ambient and occupational environment, and exposure to such materials is a well recognized cause of some lung disease. To investigate the interaction of inhaled inorganic particulates with the lung it is necessary to obtain quantitative information on the particulate burden of lung tissue in a wide variety of situations. The vast majority of diagnostic and experimental tissue samples (biopsies and autopsies) are fixed with formaldehyde solutions, dehydrated with organic solvents and embedded in paraffin wax. Over the past 16 years, I have attempted to obtain maximal analytical use of such tissue with minimal preparative steps. Unique diagnostic and research data result from both qualitative and quantitative analyses of sections. Most of the data has been related to inhaled inorganic particulates in lungs, but the basic methods are applicable to any tissues. The preparations are primarily designed for SEM use, but they are stable for storage and transport to other laboratories and several other instruments (e.g., for SIMS techniques).

Oligodendrocytes in Developing Hameter Cerebral Cortex

1976 ◽  
Vol 34 ◽  
pp. 126-127
Author(s):  
MB. Tank Buschmann
Keyword(s):  
Cerebral Cortex ◽  
Optic Nerve ◽  
Frontal Cortex ◽  
Osmium Tetroxide ◽  
Sodium Cacodylate Buffer ◽  
Sodium Cacodylate ◽  
Tissue Samples ◽  
Cacodylate Buffer ◽  
Layer V ◽  
Adult Hamster

Development of oligodendrocytes in rat corpus callosum was described as a sequential change in cytoplasmic density which progressed from light to medium to dark (1). In rat optic nerve, changes in cytoplasmic density were not observed, but significant changes in morphology occurred just prior to and during myelination (2). In our study, the ultrastructural development of oligodendrocytes was studied in newborn, 5-, 10-, 15-, 20-day and adult frontal cortex of the golden hamster (Mesocricetus auratus).Young and adult hamster brains were perfused with paraformaldehyde-glutaraldehyde in sodium cacodylate buffer at pH 7.3 according to the method of Peters (3). Tissue samples of layer V of the frontal cortex were post-fixed in 2% osmium tetroxide, dehydrated in acetone and embedded in Epon-Araldite resin.

Confocal laser microscopy of impregnated central nervous system tissue

1988 ◽  
Vol 46 ◽  
pp. 94-95
Author(s):  
J.N. Turner ◽  
M. Siemens ◽  
D. Szarowski ◽  
D.N. Collins
Keyword(s):  
Electron Microscopy ◽  
Central Nervous System ◽  
Nervous System ◽  
Three Dimensional ◽  
Dimensional Structure ◽  
Confocal Laser ◽  
High Contrast ◽  
Central Nervous System Tissue ◽  
Tissue Samples ◽  
Reflected Light

A classic preparation of central nervous system tissue (CNS) is the Golgi procedure popularized by Cajal. The method is partially specific as only a few cells are impregnated with silver chromate usualy after osmium post fixation. Samples are observable by light (LM) or electron microscopy (EM). However, the impregnation is often so dense that structures are masked in EM, and the osmium background may be undesirable in LM. Gold toning is used for a subtle but high contrast EM preparation, and osmium can be omitted for LM. We are investigating these preparations as part of a study to develop correlative LM and EM (particularly HVEM) methodologies in neurobiology. Confocal light microscopy is particularly useful as the impregnated cells have extensive three-dimensional structure in tissue samples from one to several hundred micrometers thick. Boyde has observed similar preparations in the tandem scanning reflected light microscope (TSRLM).

An Electron Microscopic Study of the Acute Effects of Hypothalamic Releasing Factors on the Anterior Pituitary Gland

1968 ◽  
Vol 26 ◽  
pp. 232-233
Author(s):  
P.W. Coates ◽  
E.A. Ashby ◽  
L. Krulich ◽  
A. Dhariwal ◽  
S. McCann
Keyword(s):  
Anterior Pituitary ◽  
Microscopic Investigation ◽  
Uranyl Acetate ◽  
Electron Microscopic Investigation ◽  
Electron Microscopic ◽  
Tissue Samples ◽  
Thin Sections ◽  
Dense Membrane ◽  
Membrane Bound ◽  
Releasing Factors

The morphologic effects on somatotrophs of crude sheep hypothalamic extract prepared from stalk-median eminence were studied by electron microscopy in conjunction with concurrently run bioassays performed on the same tissue samples taken from young adult male Sherman rats.Groups were divided into uninjected controls and injected experimentals sacrificed at 5', 15', and 30' after injection. Half of each anterior pituitary was prepared for electron microscopic investigation, the other half for bioassay. Fixation using collidine buffered osmium tetroxide was followed by dehydration and embedment in Maraglas. Uranyl acetate and lead citrate were used as stains. Thin sections were examined in a Philips EM 200.Somatotrophs from uninjected controls appeared as described in the literature (Fig. 1). In addition to other components, these cells contained moderate numbers of spherical, electron-dense, membrane-bound granules approximately 350 millicrons in diameter.

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