瑞柏 ND-Histology Products

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瑞柏 ND-Histology Products

  1. 1. Histology 組織學相關產品 + 一點閃爍計數液 Edith 2012.02.04
  2. 2. 切片冷凍切片 frozen sections 急速冷凍▸切片▸染色 固定▸脫水▸包埋▸切片▸染色石蠟切片 paraffin-embedded sections 固定▸脫鈣▸脫水▸澄清▸浸潤▸包埋▸切片▸染色
  3. 3. 固定 Fixation保持細胞、組織的固有形態和結構使原來的軟組織硬化,而使得隨後的操作程序容易進行常使用 福馬林相關溶液ex. 中性福馬林 (10% Buffered neutral formalin) 溴化氨福馬林溶液 (Formalin Ammonium Bromide Solution ; FAB) 福馬林-酒精-醋酸溶液 (Formalin- Alcohol-Acetic Acid Solution; FAA)
  4. 4. Mirskys Fixative HS-101 & HS-102冷凍切片 / 石蠟切片Saponin( 皂素 )–Glyoxal( 乙二醛 ) type fixative相較於 formalin: o reduced toxicity o superior preservation of epitopes for IHC & ICC o improved retention of tissue morphology x tissue blocks < 3mm
  5. 5. 脫鈣 Decalcification骨組織及鈣化病灶,經過固定後,必先將鈣鹽除去使組織軟化,才能進行常規切片。如脫鈣不全則切片易撕開或碎裂並損傷切片刀刃。Mineral Acids 礦物酸 / 無機酸 (ex. Nitric Acid, Hydrochloric Acid) 作用快 ; 適用於質密的骨 sample Damage Cellular MorphologyOrganic Acids 有機酸 (ex. Acetic Acid, Formic Acid) 作用較慢 ; 適用於 bone marrow, other soft tissuesEDTA 作用最慢 不適合常規使用 增加脫鈣速度則犧牲細胞微細構造
  6. 6. HS-104Calci-Clear HS-105Calci-Clear RapidModerate speed decalcifier.▸ Mildness balanced with performance▸ Minimized loss of tissue structure▸ Excellent staining characteristics 增加脫鈣速度則犧牲細胞微細構造
  7. 7. 脫水 Dehydration澄清 Clearing將已固定組織內之游離水份脫除,便於無法與脫水劑 , 水互溶之包埋劑浸潤乙醇 HS-300 (95% Ethanol/5% Methanol)Xylene 二甲苯 Histo-Clear Histo-Clear II
  8. 8. HS-200Histo-Clear HS-202Histo-Clear II無毒 + Biodegradable 使用 Histo-Clear 的客戶較多 成份略不同 Histo-Clear II 較無侵蝕性 , 但用以替換 xylene 可能需 較長的 incubation time Histo-Clear II 需與特定的 mounting medium 搭配使用 , ex. Omnimount(HS-106) HS-100 Histosol Flash point 較 xylene 高
  9. 9. 浸潤 Infiltration包埋 Embedding 包埋劑呈液態時可經由置換作用 而進入組織間隙 , 爾後凝固以便 切片 常用之包埋劑 : 石蠟 (Paraffin) 賽勞定 (Celloidin)
  10. 10. 切片 Sectioning石臘旋轉 15-25μm式 ↓切 3-5μm片機
  11. 11. 染色 StainingAntigen RetrievalProtocol 裡的xylene 亦可用Histo-Clear 取代 !
  12. 12. 染劑 Stains HS-504Alcian Blueacid mucopolysaccharides 酸性粘多醣體 ; mast cell differentiation HS-518Basic Fuchsin 鹼性品紅 / 鹼性復紅 / 鹼性洋紅Main ingredient of Schiffs reagent, a pH indicator, and stain forglycoproteins and mucopolysaccharide proteins. HS-506Biebrich Scarlet 猩紅質Plasma stain substituted for Acid Fuchsin in Massons trichrome. HS-516Fast Green FCFFibre stain substituted for Light Green SF Yellowish in Massons trichrome.
  13. 13. 染劑 Stains HS-603Bromophenol BlueCommonly used tracking dye in electrophoresis. HS-601Amido Black法醫學染色:強化微弱血跡 HS-606Methyl Green 甲 綠 基In conjunction with Pyronin Y will differentiate RNA and DNA. HS-607Pyronin Y #61In conjunction with Methyl Green will differentiate RNA and DNA. HS-525 Methylene Blue 亞 藍 甲 A sensitive stain specific for DNA and RNA.
  14. 14. 封片 Mounting
  15. 15. 封片 Mounting封片膠 / 封蓋劑 ex. 阿拉伯膠、加拿大膠 ...Mounting Media ─ aqueous ─ non-aqueous/organic先將滴一滴封片膠在組織中央,已 45 度角緩緩將蓋玻片蓋上,再以鑷子輕壓將氣泡趕出
  16. 16. HS-103HistomountClassic choice in synthetic mounting media HS-106HydromountNonfluorescing aqueous mounting mediumEffective for frozen sections, amyloid, and immunofluorescent stainingproceduresMay be removed by soaking the slides in warm saline HS-110OmnimountUniversally compatible synthetic mounting medium. Ideally suited foruse with HistoClear II.Higher flash point and Lower toxicity than xylene based mountantsProvides both a safer work environment and reduces shipping costs
  17. 17. 順便一併提一下 ... Anti-Fade* 不是所有 Mounting Solution 都含有 Anti-Fade!* ReadiUse™ Microscope Mounting Solution ( 編號 20009) FluoroQuest™ Anti-fading Kit I *for Slide* ( 編號 20001) FluoroQuest™ Anti-fading Kit II *for Plate* ( 編號 20003) AAT : 20009 不含 Anti-Fade 成份 Fluoromount( 編號 NB900-66726)( 含 Anti-Fade) We do not add anti-fading compounds to our media in order to allow the maximum flexibility in their use, but they are compatible with all the common anti-fade additives- your customer can add, for example, DABCO to Hydromount, and achieve excellent result. i-BRITE Plus( 編號 SF40000-10)( 含 Anti-Fade) Mounting Medium, Aqueous, BioAssay™ ( 編號 M4679-01) Mounting Medium, Organic, BioAssay™ ( 編號 M4679) USB : There is no antifade added to either M4679 or M4679-01.
  18. 18. 參考連結 Tissue Preparation for Light Microscopy http://www.youtube.com/watch?v=TLm37BbR1moProtocols Online | Archive | Histologyhttp://protocolsonline.com/category/histology/獸醫科技資訊網:參考手冊 動物傳染病實驗室檢驗方法 總論 第 二   十 章病 學 查   http://vettech.nvri.gov.tw/Articles/handbook/916.html 理 檢 法 Rotary Microtome Section http://www.youtube.com/watch?v=KnMdSgd5mts Immunohistochemistry http://www.youtube.com/watch?v=mpK5ojrXQs0
  19. 19. 液相閃爍計數 LSCLiquid Scintillation Counting閃爍計數器為利用某些物質所產生的閃爍光來偵測游離輻射其檢體測試形式可分為液體或固體液體閃爍計數器是利用閃爍材料如螢光劑將放射性同位素之輻射能轉變為光能再由光電管將光能轉為電脈衝進而測定輻射線的數量及能量 成大 - 藥理學研究所 - 公用儀器 -B- 計數器說明 http://www.ncku.edu.tw/~pharma/v2011/8-2.php
  20. 20. 液相閃爍計數 LSCLiquid Scintillation Counting Cocktails ─ 1.Solvent 溶解度 collector & conductor of energy must not quench Scintillator 2.Scintillator - primary 能被 excited solvent molecules 激發 - secondary wavelength shifters( 早期 ) improve efficiency( 現在 )
  21. 21. 液相閃爍計數 LSCLiquid Scintillation Counting
  22. 22. 液相閃爍計數 LSCLiquid Scintillation Counting- Metabolic studies- Protein and DNA analysis studies 細胞增生實驗 (3H-Thymidine Incorporation/Cell Proliferation Assay)利用活細胞合成 DNA 的過程,使其攝入放射性 3H-Thymidine , 依其放射強度代表細胞數目多寡一排一排地將細胞 收集 (Cell Harvester) 至濾紙 ( 玻璃纖維 ) 上,然後將含有 DNA 的濾紙剪下,置入計測瓶內,每個計測瓶加 1.5 - 2 ml 閃爍計數液,置入 Beta counter 內計測
  23. 23. 液相閃爍計數 LSCLiquid Scintillation Counting細胞收集器(Cell Harvester)
  24. 24. 液相閃爍計數 LSC Liquid Scintillation Counting液態閃爍計數器(Liquid Scintillation Analyzer, β-Counter) 國立陽明大學 高雄醫學大學 臺灣大學 國衛院育成中心 中興大學 東海大學 中山醫學大學 中臺科技大學 屏東科技大學 ...
  25. 25. 液相閃爍計數 LSCLiquid Scintillation CountingScintillation Vials Precision Molded Highest Quality HDPE Leak-Tight Seal Economical SVC-20 20 ml SVC-08 8 ml SVC-06 6 ml
  26. 26. 閃爍計數液 LSC CocktailsThe Waste Disposal Issues      in Scintillation Counting The components of the LSC cocktail may represent a hazard or a disposal problem in addition to the radioactivity. For many experiments, only a small percentage of the samples counted will have significant radioactivity, so disposal of the LSC is the predominating issue. Fortunately, biodegradable LSC cocktails have been developed, such as N.D.s Ecoscint fluids and Uniscint BD, which substantially reduce the difficulty of disposing of LSC waste.
  27. 27. 閃爍計數液 LSC Cocktails LS-273 Ecoscint A The Ideal Universal Scintillation Cocktail All Around Performance High Efficiency with High Sample Holding Biodegradable, Reduced Toxicity Solvent Biodegradable High Flash Point (>150°C) Exce ent for: ll Agar e gels, aqueous sa os mples, biologica samples, environmental samples, glass l fibe or paper filters plasma, polyacry mide gels, salt solutions, sucrose, r , la solu ized tissues. bl Useful for : Acidic sam es, alkaline sam es, catecholamine assays, high pl pl salt solut ns, tissue homog ates. io en
  28. 28. Fin.

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