Related Experiment Video
Updated: Jul 29, 2026
11:53
Immunodetection of Outer Membrane Proteins by Flow Cytometry of Isolated Mitochondria
Published on: September 18, 2014
15.5K
An algorithm-based technique for counting mitochondria in cells using immunohistochemical staining of formalin-fixed
Mai Sakashita1,2, Noriko Motoi2,3, Gaku Yamamoto4
1Division of Biomarker Discovery, Exploratory Oncology Research and Clinical Trial Center, National Cancer Center, Chiba, Japan.
Journal of Cancer Research and Clinical Oncology
|April 2, 2024
Summary
We developed an objective Mito-score system to quantify mitochondria in cancer cells using immunohistochemistry. This novel method aids in understanding cancer biology and metabolic status from pathological specimens.
Area of Science:
- * Cancer Biology
- * Cellular Metabolism
- * Immunohistochemistry
Background:
- * Visualizing mitochondria in human cancer cells is crucial for understanding cancer biology.
- * Current methods for evaluating mitochondria via immunohistochemistry are challenging due to ubiquitous mitochondrial presence and variable numbers per cell.
- * Objective quantification of mitochondrial status is needed for pathological specimens.
Purpose of the Study:
- * To establish an objective system, the Mito-score, for evaluating mitochondria in cancer cells.
- * To enable machine-based processing of hue, saturation, and value color spaces for mitochondrial analysis.
- * To provide insights into the metabolic status of cancer through mitochondrial evaluation.
Main Methods:
- * The Mito-score was defined as the ratio of COX4-positive pixels (mitochondrial marker) to the number of nuclei per cell.
- * The system was validated using lung cancer cell lines, normal tissues, and 199 lung cancer tissue specimens.
- * Machine-based processing of color spaces (hue, saturation, value) was employed for quantification.
Main Results:
- * The Mito-score demonstrated strong correlations with MitoTracker staining (R² = 0.68) and electron microscopy counts (R² = 0.79).
- * Significant differences in Mito-score were observed among lung cancer subtypes: adenocarcinoma (147.5) > squamous cell carcinoma (120.6) > large cell neuroendocrine carcinoma (111.8) > small cell carcinoma (57.25).
- * The Mito-score effectively quantifies mitochondrial content in human formalin-fixed paraffin-embedded specimens.
Conclusions:
- * The Mito-score provides a reliable and objective method for analyzing mitochondrial status in pathological cancer specimens.
- * This approach facilitates the study of mitochondrial function and its role in cancer metabolism.
- * The Mito-score has the potential to enhance our understanding of cancer biology and inform clinical insights.
Related Concept Videos
01:23Flow Cytometry
The development of flow cytometry techniques began in 1934 with initial attempts by Andrew Moldavan, a bacteriologist who counted the cells in a flowing capillary system. Moldavan pumped cells through a capillary tube focused under a microscope for visualization. The invention of photometry allowed the measurement of differentially-stained cells, and Louis Kamentsky developed the first multiparameter flow cytometer in 1965 to identify and count the cancer cells in cervical tissue specimens.
In...
In...
01:03Fixation and Sectioning
Two basic types of preparation are used to visualize specimens with a light microscope: wet mounts and fixed specimens.
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
The simplest type of preparation is the wet mount, in which the specimen is placed in a drop of liquid on the slide. A liquid specimen can be directly deposited on the slide using a dropper. Solid specimens, such as skin scraping, can be placed on the slide before adding a drop of liquid to prepare the wet mount. Sometimes the liquid is simply water, but stains are often added...
01:20Immunogold Electron Microscopy
Immunoelectron microscopy utilizes immunogold labeling of endogenous proteins with specific antibodies to detect and localize these proteins in cells and tissues. The procedure provides insights into the distribution and quantification of protein under different stimulation conditions offering clues about their functions. Conjugating highly electron-dense gold particles with primary or secondary antibodies allow antigen detection on and within cells, with high resolution and specificity.
01:13Special Staining Techniques
Specialized staining techniques play a vital role in microbiology by enabling the visualization of specific bacterial structures that remain undetectable with standard microscopy methods. These techniques not only enhance the structural visualization of bacterial cells but also provide critical insights into their pathogenicity and classification. Additionally, they support diagnostic and research endeavors in microbiology by identifying key bacterial features.Capsule Staining for Virulence...