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標題:相位差顯微鏡加強光與暗的對照可鑒別觀察物

信息分類:站內(nèi)新聞   作者:yiyi發(fā)布   時間:2011-10-30 13:36:31 將本頁加入收藏

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我讀原文書看到光學顯微鏡的部分時書上介紹很多種

像"bright field microscopy"

"phase contrast microscopy"

"differential interference contrast microscopy"

....等等

但是我剛剛看網(wǎng)路上介紹光學顯微鏡
只有複式和解剖這兩種光學
可以幫我解釋上述這幾種顯微鏡嗎???
看不太懂原文寫什么

北京上光儀器回覆:
bright feild microscopy(亮視野顯微鏡)就是我們平常使用的顯微鏡,複式和解剖顯微鏡都是。phase contrast microscopy(相位差顯微鏡)利用物體的顏色和吸光性的不同,也就是加強光與暗的對照,使可鑑別顯微鏡下的觀察物,可用于觀察活體細胞。 differential interference contrast microscopy,一如相位差顯微鏡,利用光學特徵的不同,加強因密度而產(chǎn)生光與影的效果。


我想要了解的是這篇文章到底在說些啥?!
螢光顯微鏡 (的期刊的生物化學 VOL.283、 號 4 頁 2454–2464) 后轉(zhuǎn)染,儲存格被沖洗磷酸鹽緩沖鹽和固定在 5 分鐘 25 C 后, 跟在 15 分鐘跌 C 的甲醇磷酸鹽緩沖鹽水的 4%甲醛。 固定的儲存格被探測兔抗 hNinein 血清 (1:500,我們準備)、 兔抗 Astrin 抗體我們製備 1:500)、 滑鼠抗 Astrin 抗體 (1:500,我們製備) 兔 anti-Aurora 一 (1:500,我們製備),兔 antipericentrin (1:500,Abcam) 滑鼠抗-BubR1 (1:250,屋宇署生物科學) 滑鼠抗 CENP-E (1:250,Abcam) 滑鼠 anti-tubulin 抗體 (1000 ; Sigma GTU 九八七) 和 (1000 ; 糖尿病 1A,Sigma) 的滑鼠 anti-tubulin 抗體。 DNA 是 4,6-diamidino-2-phenylindole (2 μg/毫升) 染色。 螢光細胞圖像被獲得一個顯微鏡 (Olympus) 的 Olympus LSM Fluoview 焦 500 鐳射。
Adobe Photoshop 軟體 (Adobe Systems) 處理影像。 為了避免潛在的差異,由于褪色的同一個會話過程中收集所有的資料。 圖像,然后測量的每個圖像的地區(qū)和總圖元數(shù) circumscribing 主軸輪廓并選擇該區(qū)域的定量。 圖元數(shù)獲得通過將選定的區(qū)域乘以圖元強度。
Immunofluorescence Microscopy (THE JOURNAL OF BIOLOGICAL
CHEMISTRY VOL. 283, NO. 4, pp. 2454–2464)
After transfection, the cells were washed with phosphate-buffered saline and fixed with
4% formaldehyde in phosphate-buffered saline for 5 min at 25 °C followed by methanol
at -20 °C for 15 min. The fixed cells were probed with rabbit anti-hNinein serum (1:500,
our preparation), rabbit anti-Astrin antibody (1:500, our preparation), mouse anti-Astrin
antibody (1:500, our preparation), rabbit anti-Aurora A (1:500, our preparation), rabbit
antipericentrin (1:500, Abcam), mouse anti-BubR1 (1:250, BD Biosciences), mouse
anti-CENP-E (1:250, Abcam), mouse anti-tubulin antibody (1:1000; GTU-88, Sigma),
and mouse anti-tubulin antibody (1:1000; DM 1A, Sigma). DNA was stained with
4,6-diamidino-2-phenylindole (2 μg/ml). Immunofluorescent cell images were acquired
using an Olympus LSM Fluoview 500 Confocal laser scanning microscope (Olympus).
Images were processed with Adobe Photoshop software (Adobe Systems). All data were
collected during the same session to avoid potential differences due to fading. Images
were then measured for area and total pixel number for each image by circumscribing
spindle contours and selecting that region for quantitation. Pixel number was obtained by
multiplying the selected area by pixel intensity.











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