• 한국응용곤충학회지
• /
• 제23권4호
• /
• pp.215-220
• /
• 1984

Dienes 염색법을 국내에 발생하는 마이코플라스마성 식물병의 진단에 적용해 보면서 본 염색법의 보다 효과적인 이용을 위한 실험에서 얻어진 결과를 요약하면 다음과 같다. 1 Dienes 염색법을을 통한 식물 마이코플라스마병의 진단방법은 일일초와 뽕나무에서만 지정적이었고, 대추나무, 오동나무 및 붉나무에서는 부정적이었다. 2. Dienes 염색액의 $0.2\%$와 $0.4\%$ 수용액이 일일초와 뽕나무에 각각 적합했으며, 염색시간은 $5\~10$분 그리고 탈색시간은 $10\~20$분이 좋았다. 3. 정확한 현미경적 관찰이나 선명한 사진을 위해서 절편의 두께는 $50\~100{\mu}$가 적당하였다. 4. 공시식물의 각 부위중에서 줄기 정부 바로 밑의 부위가 좋은 결과를 나타냈으며, 엽병이나 중륵도 거의 동등한 결과를 나타냈다. 5. 이병식물에서 염색반응의 정도는 병세와 비례했으며, 병징발견 초기의 시료는 건전시료와 염색반응에 있어서 쉽게 구별되지 않았다. 6. 시료를 종절할 경우 대부분의 관요소들이 상처를 받지 않을 뿐만 아니라 정확한 관찰이 용역하므로 시료를 횡절할 경우보다 효과적이었다. 7. 염색된 시료를 현미경 관찰할 경우 filter를 사용하지 않고 텅스텐 적색광을 그대로 사용하면 사관요소와 도관요소 간의 색갈 대비를 증가시키며 정확한 해석에 도움을 주었다. 8. 시료를 고정액에 고정시키면 염색반응이 방해되었으며, 시료는 신선한 것일수록 염색반응이 뚜렷하였다.

• 한국버섯학회지
• /
• 제10권1호
• /
• pp.44-48
• /
• 2012

본 연구에서는 배양된 액체종균의 세균 오염 여부를 Giemsa 용액의 단일 염색으로 짧은 시간 안에 정확하게 판정이 가능한지를 조사하였다. Giemsa 용액은 혈액 골수 림프절 말라리아 원충 리케차 세포 등을 염색하는 것으로, 염기성 색소인 methylene azul과 methylene blue, 그리고 산성색소인 eosine을 methyl alcohol-glycerine에 녹여서 제조하였다. 그리고 팽이 액체종균의 폭기 배양액 분취하여 슬라이드에 올리고 Gimesa 용액으로 염색한 후 광학 현미경으로 검사하였다. 이 결과 40~60초 동안의 세균세포를 (팽나무)버섯(류) 균사세포의 부스러기나 잔존 대두박 등과 구별할 수 있었다. 이 Gimesa 용액을 이용한 염색 및 검경방법은 빠르고 간편하며 정확하므로 액체종균을 사용하는 버섯재배 농가에서도 세균 오염을 효과적으로 동정(감지)하는데 이용할 수 있다고 생각한다.

• Clinical and Experimental Reproductive Medicine
• /
• 제28권1호
• /
• pp.13-24
• /
• 2001

Objective: In muscle and neuronal cells, calcium channels have been classified by electrophysiological and pharmacological properties into (1) voltage-dependent $Ca^{2+}$-channel (1) P/Q-type $Ca^{2+}$-channel (2) N-type $Ca^{2+}$-channel (3) L-type $Ca^{2+}$-channel (4) T-type $Ca^{2+}$-channel (5) R-type $Ca^{2+}$-channel. The present study was done in order to investigate whether there is any difference in $Ca^{2+}$-channel distribution between activated and normally fertilized embryos. Methods: The immunocytochemical method was used to identify the existence of voltage-dependent $Ca^{2+}$-channels in parthenogenetically activated 2-cell embryos by ethanol and $SrCl_2$ treatment. These 2-cell embryos were obtained by exposure to 6% ethanol for 6 min and to 10 mM $SrCl_2$ for 2h. Results: P/Q-type $Ca^{2+}$-channels and L-type $Ca^{2+}$-channels have been identified. Whereas, three type of $Ca^{2+}$-channel P/Q-type, N-type, L-type have been identified in 2-cell embryos fertilized in vivo. Conclusion: Activation by ethanol was faster than those by $SrCl_2$. However, there was difference in DAB staining of the embryos between ethanol and $SrCl_2$ treatment (87.7% and 54.1 %). Intensity of staining was also different between ethanol- and $SrCl_2$-treated group. However, it has not been known why there was some difference in DAB staining and staining intensity in the present study.

• Asian-Australasian Journal of Animal Sciences
• /
• 제18권5호
• /
• pp.626-631
• /
• 2005

Nitric oxide (NO) derived from inducible nitric oxide synthase (iNOS) is involved in cell apoptosis, which contributes to luteal regression and luteolysis in some species. In large domestic animals, no direct evidence for the relationship between NO and cell apoptosis in the process of corpus luteum regression is reported. The present study was conducted to investigate the localization of iNOS on porcine corpora lutea (CL) during the oestrus cycle and its relation to cell DNA fragmentation and CL regression. According to morphology, four luteal phases throughout the estrous cycle were defined as CL1, CL2, CL3 and CL4. By isoform-specific antibody against iNOS, the immunochemial staining was determined. Luteal cell DNA fragmentation was determined by flow cytometry. The results showed that no positive staining for iNOS was in CL1 and that iNOS was produced but limited to the periphery of CL2, while in the CL3, the spreading immunochemical staining was found inside the CL. No iNOS positive staining was detected in CL4. Meanwhile, DNA fragmentation increased dramatically when CL developed from CL2 to CL3 (p<0.05). In CL4, higher proportion of luteal cells still had fragmented DNA than that of luteal cells from CL1 or CL2 (p<0.05). These results indicate that iNOS expression is closely related to luteal cell apoptosis and then to luteal regression.

• Asian Pacific Journal of Cancer Prevention
• /
• 제14권10호
• /
• pp.5731-5734
• /
• 2013

Background: The object of this study was to examine whether a new protocol including step-sectioning and immunohistochemistry (IHC) staining of axillary sentinel nodes (SN) would lead to detection of more metastases in patients with breast cancer. Materials and Methods: Sixty-nine tumor free sentinel lymph nodes were examined. Step frozen sectioning was performed on formalin fixed SN and stained both by hematoxylin and eosin (H and E) and cytokeratin markers using IHC. Any tumoral cell in IHC stained slides were considered as a positive result. Metastases up to 0.2 mm were considered as isolated tumor cells and 0.2 up to 2 mm as micrometastasis. Results: Mean age of the patients was $48.7{\pm}12.2$ years. Step sectioning of the SN revealed 11 involved by metastasis which was statistically significant (p<0.001). Furthermore, 15 (21.7%) of the patients revealed positive results in IHC staining for pan-CK marker and this was also statistically significant (p=0.001). Ten patients had tumoral involvement in lymph nodes harvested from axillary dissection and 4 out of 15 lymph nodes with positive result for CK marker were isolated tumor cells. However, 4 of 10 patients with tumor positive lymph nodes in axillary dissection were negative for CK marker and in contrast 6 of the pan-CK positive SN were in patients with tumor-free axillary lymph nodes. Conclusions: Both IHC and step sectioning improve the detection rate of metastases. Considering the similar power of these two methods, we recommend using either IHC staining or step sectioning for better evaluation of harvested SNs.

• Asian Pacific Journal of Cancer Prevention
• /
• 제17권12호
• /
• pp.5247-5250
• /
• 2016

Objective: Endometriosis, one of the most common estrogen dependent gynecological disorders, can present as both benign and malignant disease. The prevalence of tumoral transformation is 0.7-1.6% and the most common tumors are clear cell and endometrioid carcinomas. Unfortunately, the pathogenesis of transformation is unknown. For this purpose, we examined molecular alterations in ovarian endometriosis and endometriosis-associated tumors. Methods: Using the data bank of Alzahra hospital pathology department and paraffin blocks from appropriate cases were identified. Sections were cut and stained for 3 markers: estrogen receptor, P53 and bcl2. Correlations between findings were investigated. Results: Nineteen cases of endometriosis-associated tumor and 19 cases of endometriosis were identified. Staining for bcl2 was documented in 14 of 19 (73.7%) of endometriosis-associated tumor cases and also 7 of 19 (36.8%) endometriosis cases (P=0.02). Only 3 of the 19 (15.8%) endometriosis-associated tumors exhibited positive staining for estrogen receptors, compared with 14 of 19 (73.7%) endometriosis cases (P<0.001). Positive staining for P53 was noted in 5 of 19 (31.6%) endometriosis-associated ovarian tumor samples but was absent in endometriosis samples (0%), (P =0.008). Conclusions: Endometriosis-associated tumors appear to be associated with overexpression of bcl2 and P53 and reduced expression of Estrogen receptor. These finding may help to diagnose tumoral transformation with a background of endometriosis.

• Asian Pacific Journal of Cancer Prevention
• /
• 제16권8호
• /
• pp.3147-3152
• /
• 2015

Background: Phylloides tumors are rare breast neoplasms with a variable clinical course depending on the tumor category. Along with histologic features, the role of immunohistochemical staining has been studied in predicting their behavior. Objectives: Our aim was to evaluate the role of CD 10 immunohistochemical staining in predicting survival, recurrence and metastasis in phylloides tumor. We also evaluated correlations of other clinicopathological features with overall and disease-free survival. Materials and Methods: CD10 expression was studied in 82 phylloides tumors divided into recurrent/metastatic and non-recurrent/non-metastatic cohorts. The Chi-square test was applied to determine the significance of differences in CD10 expression between outcome cohorts. Uni and multivariate survival analyses were also performed using log-rank test and Cox regression hazard models. Results: All 3 metastatic cases, 5 out of 6 (83.3%) recurrent cases and 37out of 73 (50.7%) non-recurrent and non-metastatic cases expressed significant (2+ or 3+) staining for CD10. This expression significantly varied between outcome cohorts (p<0.03). Tumor category and histological features including mitotic count and necrosis correlated significantly with recurrence and metastasis. A significant decrease in overall and disease free survival was seen with CD10 positivity, malignant category, increased mitoses and necrosis. Neither CD10 expression nor any other clinicopathologic feature proved to be an independent prognostic indicator in multivariate analysis. Conclusions: CD10 immunohistochemical staining can be used as a predictive tool for phylloides tumor but this expression should be interpreted in conjunction with tumor category.

• Journal of Ginseng Research
• /
• 제30권3호
• /
• pp.153-157
• /
• 2006

홍삼을 습점 또는 수침처리하여 원형을 복원한 후 경흔적 기산법과 나이테염색법을 이용하여 연근을 판별하였다. 경흔적 기산법은 뇌두를 $50^{\circ}C$에서 3일간 침지하여 뇌두의 뿌리 조직을 제거하면 줄기 조직만 남게 되어 연근판별의 정확도를 기할 수 있었다. 나이테 염색법은 홍삼 절편을 만들어 기존의 safranine에 의한 나이테 염색을 시도하였으나 나이테가 염색되지 않아 연근 판별이 불가능하였다. 한편 기계건조법은 홍삼 절편을 습점 처리하여 나이테를 건조시키던가 아니면 홍삼절편을 수침 처리하여 홍삼색소를 제거한 후 건조 사키면 나이테가 선명하게 들어나 연근판별이 가능하였다.

• 대한수의학회지
• /
• 제44권4호
• /
• pp.539-549
• /
• 2004

Mean death time of inoculated embryonated egg is one of the methods to determine the virulence of the Newcastle disease viruses (NDV). Evaluation of tissue tropism of NDV in the host has been proposed as an another way to determine the pathogenicity of NDV based on the principal site of viral replication. To evaluate the tissue tropism among NDV, an immunohistochemistry(IHC) technique using monoclonal antibody was applied in one-day-old SPF chickens inoculated with different ND vaccine strains such as Ulster 2C, VG/GA and B1 viruses by eye drop instillation. The tissues used for this comparison were trachea, intestine, Harderian gland and cecal tonsil, which were collected at 0.5, 1, 2, 3, 5, 8, 10, 14 days post inoculation. Among test groups, chickens inoculated with B1 viurs, which is known to replicate in the respiratory system, have IHC positive staining mainly in the trachea and those inoculated with Ulster 2C have IHC positive staining mainly in the intestine. However, chickens inoculated with VG/GA strain have IHC positive staining in both the trachea and intestine. Therefore, a differences in tissue tropism among ND vaccine strains has been proved by the IHC technique. Based on this results, the IHC staining technique could be used to classify the NDV or to study the pathogenesis of NDV in chickens.

• 치위생과학회지
• /
• 제21권4호
• /
• pp.219-226
• /
• 2021

Background: Proper detection and management of dental plaque are essential for individual oral health. We aimed to evaluate the maturation level of dental plaque using a two-tone disclosing agent and to compare it with the fluorescence of dental plaque on the quantitative light-induced fluorescence (QLF) image to obtain primary data for the development of a new dental plaque scoring system. Methods: Twenty-eight subjects who consented to participate after understanding the purpose of the study were screened. The images of the anterior teeth were obtained using the QLF device. Subsequently, dental plaque was stained with a two-tone disclosing solution and a photograph was obtained with a digital single-lens reflex (DSLR) camera. The staining scores were assigned as follows: 0 for no staining, 1 for pink staining, and 2 for blue staining. The marked points on the DSLR images were selected for RGB color analysis. The relationship between dental plaque maturation and the red/green (R/G) ratio was evaluated using Spearman's rank correlation. Additionally, different red fluorescence values according to dental plaque accumulation were assessed using one-way analysis of variance followed by Scheffe's post-hoc test to identify statistically significant differences between the groups. Results: A comparison of the intensity of red fluorescence according to the maturation of the two-tone stained dental plaque confirmed that R/G ratio was higher in the QLF images with dental plaque maturation (p<0.001). Correlation analysis between the stained dental plaque and the red fluorescence intensity in the QLF image confirmed an excellent positive correlation (p<0.001). Conclusion: A new plaque scoring system can be developed based on the results of the present study. In addition, these study results may also help in dental plaque management in the clinical setting.