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Preparation and observation of fresh-frozen sections of the green fluorescent protein transgenic mouse head.

Tada M, Shinohara Y, Kato I, Hiraga K, Aizawa T, Demura M, Mori Y, Shinoda H, Mizuguchi M, Kawano K - Acta Histochem Cytochem (2006)

Bottom Line: This paper describes a method of preparating of frozen mouse head sections so as to clearly observe the nature of the constituent proteins.Frozen sections of various green fluorescent protein (GFP) transgenic mouse heads were prepared using the film method developed by Kawamoto and Shimizu.This new method is suitable for observing GFP marked cells, enabling us to follow the transplanted GFP marked cells within frozen head sections.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Toyama 930-0194, Japan.

ABSTRACT
Hard tissue decalcification can cause variation in the constituent protein characteristics. This paper describes a method of preparating of frozen mouse head sections so as to clearly observe the nature of the constituent proteins. Frozen sections of various green fluorescent protein (GFP) transgenic mouse heads were prepared using the film method developed by Kawamoto and Shimizu. This method made specimen dissection without decalcification possible, wherein GFP was clearly observed in an undamaged state. Conversely, using the same method with decalcification made GFP observation in the transgenic mouse head difficult. This new method is suitable for observing GFP marked cells, enabling us to follow the transplanted GFP marked cells within frozen head sections.

No MeSH data available.


Related in: MedlinePlus

GFP mouse head sections prepared using the film method. 20-µm-thick sections of 14-week-old GFP mouse, ×20. (A) Fluorescence in a GFP mouse head section prepared by the film method. (B) Light microscopy of a hematoxylin-eosin stained GFP mouse head section prepared using the film method.
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Figure 1: GFP mouse head sections prepared using the film method. 20-µm-thick sections of 14-week-old GFP mouse, ×20. (A) Fluorescence in a GFP mouse head section prepared by the film method. (B) Light microscopy of a hematoxylin-eosin stained GFP mouse head section prepared using the film method.

Mentions: To observe the GFP fluorescence of mouse head sections, the sections were prepared as described above excluding the staining step (Figs. 1A, 2B and 2D). The film containing sample was removed from the glass slide and mounted again on another new slide to make the sample side close to the slide with 30% glycerin. Finally, the film was covered with a glass coverslip using Entellan new (MERCK, Germany) to prevent the film from wrinkling. GFP fluorescence was observed under a fluorescence microscope (Olympus, Japan). The GFP excitation beam wavelengths were between 470 and 490 nm and an emission filter, which permits the passage of 510 nm wavelengths, were used for observation.


Preparation and observation of fresh-frozen sections of the green fluorescent protein transgenic mouse head.

Tada M, Shinohara Y, Kato I, Hiraga K, Aizawa T, Demura M, Mori Y, Shinoda H, Mizuguchi M, Kawano K - Acta Histochem Cytochem (2006)

GFP mouse head sections prepared using the film method. 20-µm-thick sections of 14-week-old GFP mouse, ×20. (A) Fluorescence in a GFP mouse head section prepared by the film method. (B) Light microscopy of a hematoxylin-eosin stained GFP mouse head section prepared using the film method.
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC1828081&req=5

Figure 1: GFP mouse head sections prepared using the film method. 20-µm-thick sections of 14-week-old GFP mouse, ×20. (A) Fluorescence in a GFP mouse head section prepared by the film method. (B) Light microscopy of a hematoxylin-eosin stained GFP mouse head section prepared using the film method.
Mentions: To observe the GFP fluorescence of mouse head sections, the sections were prepared as described above excluding the staining step (Figs. 1A, 2B and 2D). The film containing sample was removed from the glass slide and mounted again on another new slide to make the sample side close to the slide with 30% glycerin. Finally, the film was covered with a glass coverslip using Entellan new (MERCK, Germany) to prevent the film from wrinkling. GFP fluorescence was observed under a fluorescence microscope (Olympus, Japan). The GFP excitation beam wavelengths were between 470 and 490 nm and an emission filter, which permits the passage of 510 nm wavelengths, were used for observation.

Bottom Line: This paper describes a method of preparating of frozen mouse head sections so as to clearly observe the nature of the constituent proteins.Frozen sections of various green fluorescent protein (GFP) transgenic mouse heads were prepared using the film method developed by Kawamoto and Shimizu.This new method is suitable for observing GFP marked cells, enabling us to follow the transplanted GFP marked cells within frozen head sections.

View Article: PubMed Central - PubMed

Affiliation: Faculty of Pharmaceutical Sciences, Toyama Medical and Pharmaceutical University, Toyama 930-0194, Japan.

ABSTRACT
Hard tissue decalcification can cause variation in the constituent protein characteristics. This paper describes a method of preparating of frozen mouse head sections so as to clearly observe the nature of the constituent proteins. Frozen sections of various green fluorescent protein (GFP) transgenic mouse heads were prepared using the film method developed by Kawamoto and Shimizu. This method made specimen dissection without decalcification possible, wherein GFP was clearly observed in an undamaged state. Conversely, using the same method with decalcification made GFP observation in the transgenic mouse head difficult. This new method is suitable for observing GFP marked cells, enabling us to follow the transplanted GFP marked cells within frozen head sections.

No MeSH data available.


Related in: MedlinePlus