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Comparison of Alcian Blue, Trypan Blue, and Toluidine Blue for Visualization of the Primo Vascular System Floating in Lymph Ducts.

Kim DU, Han JW, Jung SJ, Lee SH, Cha R, Chang BS, Soh KS - Evid Based Complement Alternat Med (2015)

Bottom Line: We found that Trypan blue and toluidine blue did not visualize the PVS.Trypan blue was cleared by the natural washing.These completely different behaviors of the three dyes were found for the first time in the current work and provide valuable information to elucidate the mechanism through which some special dyes stained the PVS preferentially compared to the lymphatic wall.

View Article: PubMed Central - PubMed

Affiliation: Nano Primo Research Center, Advanced Institute of Convergence Technology, Seoul National University, Suwon 443-270, Republic of Korea ; College of Physical Education, The University of Suwon, Hwaseong 445-743, Republic of Korea.

ABSTRACT
The primo vascular system (PVS), floating in lymph ducts, was too transparent to be observed by using a stereomicroscope. It was only detectable with the aid of staining dyes, for instance, Alcian blue, which was injected into the lymph nodes. Some dyes were absorbed preferentially by the PVS than the lymph wall. It remains a standing problem to know what dyes are absorbed better by the PVS than the lymph walls. Such information would be useful to unravel the biochemical properties of the PVS that are badly in need for obtaining large amount of PVS specimens. In the current work we tried two other familiar dyes which were used in PVS research before. We found that Trypan blue and toluidine blue did not visualize the PVS. Trypan blue was cleared by the natural washing. Toluidine blue did not stain the PVS, but it did leave stained spots in the lymph wall and its surrounding tissues, and it leaked out of the lymph wall to stain surrounding connective tissues. These completely different behaviors of the three dyes were found for the first time in the current work and provide valuable information to elucidate the mechanism through which some special dyes stained the PVS preferentially compared to the lymphatic wall.

No MeSH data available.


Related in: MedlinePlus

Rod-shaped nuclei in the primo vessel. (a) Phase contrast microscope image of a primo vessel that was extracted from a lymph duct. (b) Fluorescence microscopic image of the boxed area in (a). The boxed area was magnified, and the part with rod-shaped nuclei was the primo vessel. The round-shaped nuclei were aggregated lymphocytes. (c–f) Confocal laser scanning microscope images of the circled area in (b). These panels were consecutive optical sections from top to bottom separated by 2 μm in each step. The same rod-shaped nucleus appeared (panel (c)), looked long (panels (d) and (e)), and then nearly disappeared (panel (f)), which implies that this particular endothelial nucleus was inside the primo vessel and, therefore, had not aggregated from outside.
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fig6: Rod-shaped nuclei in the primo vessel. (a) Phase contrast microscope image of a primo vessel that was extracted from a lymph duct. (b) Fluorescence microscopic image of the boxed area in (a). The boxed area was magnified, and the part with rod-shaped nuclei was the primo vessel. The round-shaped nuclei were aggregated lymphocytes. (c–f) Confocal laser scanning microscope images of the circled area in (b). These panels were consecutive optical sections from top to bottom separated by 2 μm in each step. The same rod-shaped nucleus appeared (panel (c)), looked long (panels (d) and (e)), and then nearly disappeared (panel (f)), which implies that this particular endothelial nucleus was inside the primo vessel and, therefore, had not aggregated from outside.

Mentions: The primo node is an oval-shaped tissue packed with various cells [12]. Figure 5 shows a primo node attached to one primo vessel, during preparation. Indeed, the primo node was packed with various cell nuclei, as seen in Figure 5(b). The minimal criterion to confirm the presence of the L-PVS is the observation of a distribution of the rod-shaped nuclei by DAPI staining. Figure 5(b) shows the presence of longitudinally distributed rod-shaped nuclei; more are shown in Figure 6.


Comparison of Alcian Blue, Trypan Blue, and Toluidine Blue for Visualization of the Primo Vascular System Floating in Lymph Ducts.

Kim DU, Han JW, Jung SJ, Lee SH, Cha R, Chang BS, Soh KS - Evid Based Complement Alternat Med (2015)

Rod-shaped nuclei in the primo vessel. (a) Phase contrast microscope image of a primo vessel that was extracted from a lymph duct. (b) Fluorescence microscopic image of the boxed area in (a). The boxed area was magnified, and the part with rod-shaped nuclei was the primo vessel. The round-shaped nuclei were aggregated lymphocytes. (c–f) Confocal laser scanning microscope images of the circled area in (b). These panels were consecutive optical sections from top to bottom separated by 2 μm in each step. The same rod-shaped nucleus appeared (panel (c)), looked long (panels (d) and (e)), and then nearly disappeared (panel (f)), which implies that this particular endothelial nucleus was inside the primo vessel and, therefore, had not aggregated from outside.
© Copyright Policy - open-access
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC4562098&req=5

fig6: Rod-shaped nuclei in the primo vessel. (a) Phase contrast microscope image of a primo vessel that was extracted from a lymph duct. (b) Fluorescence microscopic image of the boxed area in (a). The boxed area was magnified, and the part with rod-shaped nuclei was the primo vessel. The round-shaped nuclei were aggregated lymphocytes. (c–f) Confocal laser scanning microscope images of the circled area in (b). These panels were consecutive optical sections from top to bottom separated by 2 μm in each step. The same rod-shaped nucleus appeared (panel (c)), looked long (panels (d) and (e)), and then nearly disappeared (panel (f)), which implies that this particular endothelial nucleus was inside the primo vessel and, therefore, had not aggregated from outside.
Mentions: The primo node is an oval-shaped tissue packed with various cells [12]. Figure 5 shows a primo node attached to one primo vessel, during preparation. Indeed, the primo node was packed with various cell nuclei, as seen in Figure 5(b). The minimal criterion to confirm the presence of the L-PVS is the observation of a distribution of the rod-shaped nuclei by DAPI staining. Figure 5(b) shows the presence of longitudinally distributed rod-shaped nuclei; more are shown in Figure 6.

Bottom Line: We found that Trypan blue and toluidine blue did not visualize the PVS.Trypan blue was cleared by the natural washing.These completely different behaviors of the three dyes were found for the first time in the current work and provide valuable information to elucidate the mechanism through which some special dyes stained the PVS preferentially compared to the lymphatic wall.

View Article: PubMed Central - PubMed

Affiliation: Nano Primo Research Center, Advanced Institute of Convergence Technology, Seoul National University, Suwon 443-270, Republic of Korea ; College of Physical Education, The University of Suwon, Hwaseong 445-743, Republic of Korea.

ABSTRACT
The primo vascular system (PVS), floating in lymph ducts, was too transparent to be observed by using a stereomicroscope. It was only detectable with the aid of staining dyes, for instance, Alcian blue, which was injected into the lymph nodes. Some dyes were absorbed preferentially by the PVS than the lymph wall. It remains a standing problem to know what dyes are absorbed better by the PVS than the lymph walls. Such information would be useful to unravel the biochemical properties of the PVS that are badly in need for obtaining large amount of PVS specimens. In the current work we tried two other familiar dyes which were used in PVS research before. We found that Trypan blue and toluidine blue did not visualize the PVS. Trypan blue was cleared by the natural washing. Toluidine blue did not stain the PVS, but it did leave stained spots in the lymph wall and its surrounding tissues, and it leaked out of the lymph wall to stain surrounding connective tissues. These completely different behaviors of the three dyes were found for the first time in the current work and provide valuable information to elucidate the mechanism through which some special dyes stained the PVS preferentially compared to the lymphatic wall.

No MeSH data available.


Related in: MedlinePlus