• Title/Summary/Keyword: cryosection

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Evaluation of Therapeutic Monitoring of Prostate Cancer (PCa) using [18F]Florastamin, Diagnostic Radiopharmaceutical for PCa: Non-clinical Ex vivo Whole-body Autoradiographic Analysis

  • Min Hwan Kim;Kyongkyu Lee;Hee Seup Kil;Soon Jeong Kwon;Yong Jin Lee;Kyo Chul Lee;Dae Yoon Chi
    • Journal of Radiopharmaceuticals and Molecular Probes
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    • v.9 no.1
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    • pp.17-21
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    • 2023
  • In this study, we evaluated the targeting of prostate cancer (PCa) using [18F]Florastamin in non-clinical study, for the purpose of therapeutic monitoring of [177Lu]Ludotadipep, a therapeutic radiopharmaceutical for PCa, [18F]Florastamin/[177Lu]Ludotadipep was co-administered to a single-individual prostate tumor bearing mouse model, mimicking clinical condition. Considering the difference in half-life of the two isotopes (18F or 177Lu), image scan of whole-body autoradiography was performed at 24 or 48 h after preparation of frozen section, respectively. Then, it was confirmed whether they showed the same targeting efficiency for the area of tumor. A tumor xenograft model was prepared using PSMA-overexpressing PC3-PIP prostate cancer cells. [18F]Florastamin [111 MBq (3 mCi) in 100 µL]/177Lu]Ludotadipep [3.7 MBq (100 µCi) in 100 µL] was co-administered through the tail vein, and 2 hours after administration, the mice were frozen, and after freezing for 24 hours, whole-body cryosection was performed at 24 h after freezing. Image scanning using cryosection was performed after 24 or 48 hours after freezing, respectively. In the scan image after 24 hours, tumor uptake of [18F] Florastamin/[177Lu]Ludotadipep were simultaneously observed specific uptake in the tumor. In the scan image after 48 hours in the same section, signal of 18F was lost by decay of radioisotope, and specific uptake image for [177Lu]Ludotadipep was observed in the tumor. Uptake of [177Lu]Ludotadipep was specific to the same tumor region where [18F]Florastamin/[177Lu]Ludotadipep was uptake. These results suggested that [18F]Florastamin showed the same tumor uptake efficiency to PCa as [177Lu]Ludotadipep, and effective therapeutic monitoring is expected to be enable using [18F]Florastamin during [177Lu]Ludotadipep therapy for PCa.

Is optimal cutting temperature compound essential embedding solution treatment to cryo-sectioning of brain tissue?

  • Baek, Hye Kyung;Song, Ji Ae;Yi, Sun Shin
    • Korean Journal of Veterinary Research
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    • v.56 no.2
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    • pp.85-89
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    • 2016
  • We tested a set of conditions for obtaining optimal tissue quality in preparation for histology in samples of mouse brain. C57BL/6J mice were sacrificed and perfused with 4% paraformaldehyde, after which the brains were removed and dehydrated in 30% sucrose solution. The brains were then divided into four groups according to freezing temperature and usage of optimal cutting temperature (OCT) compound. Next, we stained the sectioned brain tissues with Harris hematoxylin and eosin Y and immunohistochemistry was performed for doublecortin. The best quality tissue was obtained at $-25^{\circ}C$ and by not embedding with the OCT compound. When frozen at $-25^{\circ}C$, the embedded tissue was significantly damaged by crystals, while at $-80^{\circ}C$ there were no meaningful differences between qualities of embedded- and non-embedded tissues. Overall, we identified a set of conditions to obtain quality frozen brain sections. Our developed protocol will help resolve matters associated with damage caused to sectioned brain tissue by crystal formation during freezing.

Preservation of Ultrastructure of Ultrathin Frozen Sections for Immunoelectron Microscopic Observation (면역전자현미경적 관찰을 위한 동결초박절편의 미세구조 보존)

  • Kim, Yun-Sang;Chae, Hee-Sun;Kim, Kyung-Yong;Lee, Won-Bok
    • Applied Microscopy
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    • v.28 no.4
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    • pp.465-475
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    • 1998
  • The cryoprotection, section retrieval and embedding methods were studied for the preservation of ultrastructure of ultracryomicrosections in immunoelectron microscopy. The results obtained were as follows. 1. The cryoprotection of ultrastructure with a mixture containing 1.7 M sucrose and 15% polyvinylpyrrolidone was better than that with 2.3 M sucrose. The stretching caused by surface tension and the electron lucent holes decreased more in the cryosections infused with 2.3 M sucrose than in those with the mixture. 2. The difference between section retrieval solutions in cases of cryoprotection with 2.3 M sucrose was that the destructive .effects such as electron lucent holes and stretching between myofribrils were less in a mixture containing 1% methylcellulose and 2.3 M sucrose than in 2.3 M sucrose. The difference was obscure in the mixture containing 1.7 M sucrose and 15% PVP, but the destructive effects were slightly less in a mixture containing 1% mthylcellulose and 2.3 M sucrose than in 2.3 M sucrose or 1% methylcellulose. 3. The embedding of cryosection on drying with 2% PVA or 2% methylcellulose exhibited some protective effect during observation with transmission electron microscope, but made the ultrastructure more obscure. 4. Mitochondrial membrane and cristae and myofilaments were well delinated in sections infused with 2.3 M sucrose and retrieved with 1% methylcellulose and 2.3 M sucrose. In summary, it is suggested that the cryoprotection with 2.3 M sucrose and section retrieval with a mixture containing 1% methylcellulose and 2.3 M sucrose are good for the ultrastructure of cryosections.

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CHANGE OF TASTE PREFERENCE AND TASTE BUD AFTER UNILATERAL LINGUAL NERVE TRANSECTION IN RAT (백서 편측 설신경 손상 후 미각 및 설유두의 변화에 대한 연구)

  • Kim, Yoon-Tae;Jeon, Seung-Ho;Yeom, Hak-Ryol;Kang, Jin-Han;Ahn, Kang-Min;Kim, Sung-Min;Jahng, Jeong-Won;Park, Kyung-Pyo;Lee, Jong-Ho
    • Journal of the Korean Association of Oral and Maxillofacial Surgeons
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    • v.31 no.6
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    • pp.515-525
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    • 2005
  • Purpose of study: Lingual nerve damage can be caused by surgery or trauma such as physical irriatation, radiation, chemotherapy, infection and viral infection. Once nerve damage occurred, patients sometimes complain taste change and loss of taste along with serious disturbance of tongue. The purpose of this study was to evaluate the effects of unilateral lingual nerve transection on taste as well as on the maintenance of taste buds. Materials & Methods: Male Sprague-Dawley rats weighing 220-250g received unilateral transection of lingual nerve, subjected to the preference test for various taste solutions (0.1M NaCl, 0.1M sucrose, 0.01M QHCl, or 0.01M HCl) with two bottle test paradigm at 2, 4, 6, or 8 weeks after the operation. Tongue was fixed with 8% paraformaldehyde. After fixation, they were observed with scanning electron microscope(JSM-$840A^{(R)}$, JEOL, JAPAN) and counted the number of the dorsal surface of the fungiform papilla for changes of fungiform papilla. And, Fungiform papilla were obtained from coronal sections of the anterior tongue(cryosection). After cryosection, immunostaining with $G{\alpha}gust$(I-20)(Santa Cruz Biotechnology, USA), $PLC{\beta}2$(Q-15)(Santa Cruz Biotechnology, USA), and $T_1R_1$(Alpha Diagnostic International, USA) were done. Immunofluorescence of labeled taste bud cells was examined by confocal microscopy(F92-$300^{(R)}$, Olympus, JAPAN). Results: The preference score for salty and sweet tended to be higher in the operated rats with statistical significance, compared to the sham rats. Fungiform papilla counting were decreased after lingual nerve transaction. In 2 weeks, maximum differences occurred. Gustducin and $T_1R_1$ expressions of taste receptor in 2 and 4 weeks were decreased. $PLC{\beta}2$ were not expressed in both experimental and control group. Conclusion: This study demonstrated that the taste recognition for sweet and salty taste changed by week 2 and 4 after unilateral lingual nerve transection. However, regeneration related taste was occurred in the presence of preserving mesoneurial tissue and the time was 6 weeks. Our results demonstrated that unilateral lingual nerve damage caused morphological and numerical change of fungiform papilla. It should be noted in our study that lingual nerve transection resulted in not only morphological and numerical change but also functional change of fungiform papillae.

Regional Distribution of Interstitial Cells of Cajal, (ICC) in Human Stomach

  • Yun, Hyo-Yung;Sung, Ro-Hyun;Kim, Young-Chul;Choi, Woong;Kim, Hun-Sik;Kim, Heon;Lee, Gwang-Ju;You, Ra-Young;Park, Seon-Mee;Yun, Sei-Jin;Kim, Mi-Jung;Kim, Won-Seop;Song, Young-Jin;Xu, Wen-Xie;Lee, Sang-Jin
    • The Korean Journal of Physiology and Pharmacology
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    • v.14 no.5
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    • pp.317-324
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    • 2010
  • We elucidated the distribution of interstitial cells of Cajal (ICC) in human stomach, using cryosection and $c-Kit$ immunohistochemistry to identify $c-Kit$ positive ICC. Before $c-Kit$ staining, we routinely used hematoxylin and eosin (HE) staining to identify every structure of human stomach, from mucosa to longitudinal muscle. HE staining revealed that the fundus greater curvature (GC) had prominent oblique muscle layer, and $c-Kit$ immunostaining $c-Kit$ positive ICC cells were found to have typical morphology of dense fusiform cell body with multiple processes protruding from the central cell body. In particular, we could observe dense processes and ramifications of ICC in myenteric area and longitudinal muscle layer of corpus GC. Interestingly, $c-Kit$ positive ICC-like cells which had morphology very similar to ICC were found in gastric mucosa. We could not find any significant difference in the distribution of ICC between fundus and corpus, except for submucosa where the density of ICC was much higher in gastric fundus than corpus. Furthermore, there was no significant difference in the density of ICC between each area of fundus and corpus, except for muscularis mucosa. Finally, we also found similar distribution of ICC in normal and cancerous tissue obtained from a patient who underwent pancreotomy and gastrectomy. In conclusion, ICC was found ubiquitously in human stomach and the density of ICC was significantly lower in the muscularis mucosa of both fundus/corpus and higher in the submucosa of gastric fundus than corpus.