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Pinocytotic vesicles and microfilaments observed by scanning electron microscopy
Abstract:
Pinocytotic vesicles and microfilaments were clearly demonstrated by high resolution scanning electron microscopy. Mesothelial cells covering the newt liver were excised and treated with a hypotonic phosphate buffer solution (150 mOsm) for 1-2 minutes. After fixation with 2.5% glutaraldehyde, some specimens were placed in 20% glycerin solution for 24 hours. After being postfixed with 1% osmic solution, they were treated with 2% tannic acid and 1% osmic solution for conductive staining. After dehydration in a graded series of ethanol, the deeper layers of the mesothelium were exposed by scratching or rubbing the specimen in absolute ethanol. When some part of the surface cell membrane was turned over, many pinocytotic vesicles could be successfully seen attached to the cytoplasmic side of the membrane. The vesicles were also observed on the basal cell membrane. In the deeper layer of the cell exposed by scratching or rubbing the specimen, intracellular fibers presumably microfilaments were spread in the whole cell body forming "cytoskeleton".
Insights
High-resolution scanning electron microscopy revealed pinocytotic vesicles and microfilaments in newt liver mesothelial cells. This study visualizes cellular structures, offering insights into cell membrane dynamics and cytoskeleton organization.
Area of Science:
- Cell Biology
- Microscopy Techniques
Background:
- Mesothelial cells form a protective layer on organs.
- Understanding cellular structures is key to cell function.
Purpose of the Study:
- To demonstrate pinocytotic vesicles and microfilaments in newt liver mesothelial cells.
- To visualize the cytoskeleton using advanced microscopy.
Main Methods:
- High-resolution scanning electron microscopy was employed.
- Newt liver mesothelial cells were treated with hypotonic buffer and glycerin.
- Specimens underwent fixation, conductive staining, and dehydration.
- Cellular layers were exposed by scratching/rubbing in ethanol.
Main Results:
- Pinocytotic vesicles were observed on both the surface and basal cell membranes.
- Intracellular fibers, presumed microfilaments, formed a cytoskeleton throughout the cell.
- The technique allowed visualization of vesicles attached to the cytoplasmic side of the membrane.
Conclusions:
- High-resolution scanning electron microscopy effectively visualizes cellular structures like pinocytotic vesicles and microfilaments.
- The study provides detailed morphological evidence of the cytoskeleton and membrane dynamics in mesothelial cells.