Related Experiment Video
Updated: Aug 13, 2026
09:11
High-resolution Volume Imaging of Neurons by the Use of Fluorescence eXclusion Method and Dedicated Microfluidic Devices
Published on: March 26, 2018
Noninvasive measurement of cell volume changes by negative staining
Miriam S Droste1, Stefan S Biel, Lara Terstegen
1Beiersdorf AG, Research Microscopy, Hamburg, Germany and FB Naturwissenschaftliche Technik, Hochschule für Angewandte Wissenschaften, Hamburg, Germany.
Journal of Biomedical Optics
|January 18, 2006
Summary
This study introduces a novel fluorescence microscopy method using fluorescein to precisely measure cell volume changes during osmotic challenges. This technique is osmotically inert, allowing real-time monitoring of cell volume regulation and viability.
Area of Science:
- Cell Biology
- Biophysics
- Physiology
Background:
- Cells regulate internal ion and molecule concentrations against osmotic challenges using sophisticated transport systems.
- Accurate measurement of cell volume changes during osmotic stress requires osmotically inert and online monitoring techniques.
Purpose of the Study:
- To develop a simple, fluorescence microscopy-based method for monitoring cell volume changes during osmotic challenges.
- To assess cell viability concurrently with volume measurements.
Main Methods:
- Utilized fluorescein as a negative stain in a fluorescence microscopy setup.
- Applied hypo- and hyperosmotic challenges to Madin-Darby canine kidney (MDCK) cells.
- Measured cell volume changes and observed fluorescein entry into the cytoplasm.
Main Results:
- The fluorescein-based method is osmotically inert and does not affect cell membrane functionality or osmolarity.
- Precise and reproducible quantitative data on reversible cell volume changes were obtained.
- Cell viability was directly assessed by cytoplasmic stain appearance, particularly in fixed cells undergoing osmotic shock.
Conclusions:
- This fluorescence microscopy approach offers a sensitive and reliable tool for studying cell volume regulatory processes.
- The method allows for simultaneous assessment of cell volume dynamics and viability.
- It is ideal for investigating water and ion flux across cell membranes.
Related Concept Videos
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...
01:23Flow 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...