Ted Pella公司的微波技術(shù)及方法帶來了組織處理領(lǐng)域的一場(chǎng)革新。相比于傳統(tǒng)處理方法，使用新一代組織處理系統(tǒng)PELCO® BioWave® Pro+及其提供的方法，不僅組織處理時(shí)間可節(jié)省高達(dá)95%，而且能始終如一地得到的實(shí)驗(yàn)結(jié)果。
樣品溫度及微波輻射功率的控制是有效利用微波技術(shù)的關(guān)鍵。PELCO® BioWave® Pro+微波快速組織處理系統(tǒng)對(duì)溫度和樣品狀態(tài)能進(jìn)行精確的控制和監(jiān)控，所采用的*技術(shù)包括：
- 的ColdSpot™技術(shù)（美國(guó)號(hào)6329645）
- 真正的可變功率
- 數(shù)字化程序控制
- 內(nèi)置真空及水循環(huán)系統(tǒng)
- 自動(dòng)磁電管預(yù)熱
- 外置SteadyTemp™水溫控制
一、BioWave® Pro+微波處理系統(tǒng)的特點(diǎn)
1、 的微波控制功能
的微波功率和溫度控制 使樣品處理結(jié)果變得更為可靠。功率大小從110至750W之間連續(xù)可調(diào)；
2、 *的應(yīng)用靈活性
適用于電鏡組織處理、免疫標(biāo)記、脫鈣、固定等各種用途；
3、 升級(jí)的方法管理程序
提供對(duì)樣品處理方法的儲(chǔ)存、編輯、調(diào)用等各種功能；
4、 真正簡(jiǎn)便的操作過程
直觀的智能觸摸屏控制面板使得操作過程簡(jiǎn)單，所有設(shè)置一目了然；
5、 可靠的樣品處理結(jié)果
強(qiáng)大的微波控制功能保證了可靠而高質(zhì)量的樣品處理結(jié)果；
6、 精確的樣品溫度控制功能
避免了樣品受熱的不足，充分利用了電鏡樣品微波處理的優(yōu)勢(shì)。
7、 方便快捷
Pro+組織處理系統(tǒng)應(yīng)用范圍非常廣泛，只要配置相應(yīng)的處理附件，就可以在一臺(tái)設(shè)備上實(shí)現(xiàn)多種功能，主要包括：
- 電鏡組織處理
- 免疫標(biāo)記
- 甲醛固定及EDTA脫鈣
- 石蠟組織處理
- 共焦顯微鏡及原位雜交
二、新特點(diǎn)
| 友好的運(yùn)行界面 所有的參數(shù)都直觀、清晰的顯示在一個(gè)屏幕中 |  |
| 生動(dòng)的運(yùn)行-時(shí)間圖表 可迅速便捷的查看當(dāng)前執(zhí)行程序的實(shí)時(shí)運(yùn)行狀況 |  |
| 簡(jiǎn)潔的方法選擇 圖標(biāo)化的界面讓用戶瞬間就可以找到所需程序 |  |
| 全新的方法管理功能 每個(gè)用戶都可以直接在設(shè)備里創(chuàng)建自己的文件夾，用來存放所需程序，或者將程序直接導(dǎo)入U(xiǎn)盤里。因此，BioWave® Pro+非常適合中心實(shí)驗(yàn)室或多用戶實(shí)驗(yàn)室。 |  |
三、操作面板
＊ 手動(dòng)控制
＊ 自動(dòng)控制
＊ 設(shè)置
＊ 系統(tǒng)選項(xiàng)
四、技術(shù)參數(shù)
*采用*的工程技術(shù)，專為實(shí)驗(yàn)室而研制，微波發(fā)射器，*由微機(jī)控制。
*微波功率：100-750W之間連續(xù)可調(diào)，以1W遞增，微波頻率：2.45GHz
*爐腔包裹層材料防酸、防有機(jī)試劑
*時(shí)間設(shè)定：1秒到96小時(shí)之間可調(diào)
*功能控制：7＂友好直觀視窗觸摸屏人機(jī)界面，工業(yè)級(jí)觸式按鍵
*溫度控制：±1℃（對(duì)于大多數(shù)水液），溫度超出預(yù)設(shè)范圍時(shí)自動(dòng)停止微波輻射
*內(nèi)部冷卻控制：集成的水冷系統(tǒng)，循環(huán)水流量：1.5L/分鐘
*外部冷卻系統(tǒng)（選件）：PELCO Steady Temp*，450W冷卻系統(tǒng)
*磁攪拌器：內(nèi)置，0-3000rpm
*出口：110cfm容量
*放氣：門打開時(shí)自動(dòng)放氣
*大處理量：標(biāo)準(zhǔn)石蠟盒：58個(gè)，電鏡組織：48個(gè)
*樣品處理方法管理：可存儲(chǔ)多達(dá)204步的方法
*真空系統(tǒng)：20＂Hg,三種可選模式
*認(rèn)證：ETL/CE
*電壓要求：10A/230V
*尺寸：553mm x 514mm x 546mm
*重量：37kg
五、圖片效果
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Fig. 1A-C. Figure 1A is a micrograph of a normal sural nerve with a non-myelinated nerve (N) having secretory vesicles (sv), microtubules (mt) and a swan cell nucleus (ScN). The insert (1B) s a myelinated nerve and the arrows clearly demonstrate its periodicity. Figure 4C is a membranouse Lupus nehpritis (RPS/ISN Class V). There is diffuse, generalized effacement of the foot processes of the visceral epithelial cells. Numerous regularly disposed epimembraneous immune complex deposits are illustrated by the arrows. Both tissues were initially fixed in a variant of 10% NBF (Carson et al. 1972) and then processed in the microwave for ultrastructural evaluation by the methods of Giberson et al. (2003) and Austin (2002). Micrographs from Ronald L. Austin, Research Associate, Dept. of Pathology, LSU Medical Center, Shreveport, LA 71130.
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Figure 2A-G. Fig. 2A-B s cytoplasmic iridovirus from the skin of a sturgeon. The iridovirus is a large enveloped dsDNA virus which infects both insect and vertebrate hosts. Fig. 2C-E demonstrates an intranuclear baculovirus from the hepatopancreas of a crayfish from Northern California. Fig. 2C is a low magnification image of the enveloped dsDNA virus ing the intranuclear arrangement of virus particles Fig. 2D is a higher magnification ing both a cross-sectional and longitudinal view of the virus. Fig. 2E is a high magnification cross-section of a number of virus particles demonstrating the unique intranuclear membrane-bound virions. Fig. 2F-G demonstrates an endothelial cell polyoma virus from a blood vessel in the liver of a parakeet. Polyoma virus have a single molecule of circular dsDNA and the particles are non-enveloped. Polyoma virions are spherical in outline and typically 45nm in diameter. Fig. 2F is a low magnification image ing typical nuclear presentation. Fig. 2G is a high magnification view of the virus. Infected tissues were processed directly from 10% NBF by the microwave methods of Nordhausen and Barr (2001) and Nordhausen et al. (2002). Micrographs from Bob Nordhausen, Univ. of California, Davis, California Animal Health and Food Safety Lab, School of Veterinary Medicine, Davis, CA 95616.
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Figure 3. Micrographs from a 2008 microwave workshop held at the Center for Microscopy, San Joaquin Delta College, Stockton, CA. Rat brain (not perfusion fixed) (1), cardiac muscle (2), kidney (3) and liver (4) were processed from osmium through resin polymerization for a net turnaround time of under 4 hours from fresh tissue to the electron microscope. Microwave techniques (Giberson, et al., 2003) make it possible to teach and in real time demonstrate the techniques of electron microscopy.
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Figure 4A-F. Figures A-C are of normal mouse liver benched fixed in 10% NBF for 24 hours. Figures D-F are of normal mouse liver fixed in 10% NBF for 20 minutes utilizing microwave radiation. All tissues were prepared for fixation identically and cut to 2mm prior to fixation. Figures A and D are corresponding Hematoxylin and Eosin stained sections and figures B and E are corresponding Vimentin IHC stained sections. Figures C and F are corresponding EM sections demonstrating complimentary ultra-structure. Images are from Dr. Jose Galvez, Center for Comparative Medicine and Department of Medical Pathology, University of California, Davis, CA.
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Figure 5. Tissues formalin fixed and paraffin processed by the protocols described in Galvez et al. 2006. Mouse mammary tumor virus induced mammary carcinoma (A, B). Note the mitotic figures (arrows) in B. Mouse esophagus with clearly identifiable muscle striations (*) (C). Mouse uterus stained with mouse anti-estrogen (D). (Center for Comparative Medicine and Department of Pathology, Univ. of Calif., Davis)
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Figure 6: Confocal projection of Elodea canadensis labeled with Hoechst 33258 nucleic acid probe (blue stain) for 6 minutes in the absence (A) and presence (B) of 150W microwave radiation. Confocal projection of Arabidopsis thaliana root tip labeled with Hoechst 33258 nucleic acid probe after 6 minutes of 150W microwave radiation (C). Confocal projection of in situ hybridization patters of whole chromosome probes (red) hybridized to nuclei of paraffin embedded rabbit skin (D). Confocal projection of mouse kidney paraffin sections labeled with anti-Factor VIII monoclonal antibody for 60 minutes on the bench (E) and after 6 minutes of 150W radiation (F) (Scale for all bars = 50µm) (Table 1). (Mark Sanders, Imaging Center, College of Biological Sciences, Univ. of Minnesota, St. Paul, MN). Reprinted with permission of Galvez et al., Microscopy and Analysis, Nov. 2006.
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Figure 7. Retinas were fixed in 4% paraformaldehyde in 0.1M phosphate buffer (pH 7.4) overnight at 4°C. Following fixation the tissue was rinsed 6 x 20 minutes in buffer prior to beginning antibody labeling. The bench staining protocol required 7 days. The labeling results were completed in an afternoon using microwave-enhanced labeling during a workshop held at the Univ. of Minnesota Imaging Center (Mark Sanders, Director - May 17-19, 2006). The retinas were triple-labeled for: - Collagen Type IV (basal lamina surrounding blood vessels) with rabbit anti-type IV collagen and the secondary conjugated to FITC (green label)
- Glutamine Synthetase (enzyme found in retinal Müller glial cells) with mouse anti- glutamine synthetase and the secondary conjugated to Cy3 (red label)
- Glial Fibrillary acidic protein (GFAP an intermediate filament protein of astroglial cells) with chicken anti-glial fibrillary acidic protein and the secondary conjugated to Cy5 (blue label)
Primary antibody labeling was done at 170W for 12 minutes (4 on - 4 off - 4 on) under vacuum (15" Hg). Secondary antibody labeling was done at 170W for 6 minutes (2 on - 2 off - 2 on) under vacuum (15" Hg). Images were collected on a Nikon C1si Confocal Microscope.
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Figure 8. Note the symmetrical pattern of demineralization when microwave methods are employed. The separated piece at the top broke off during plastic embedding. From the work of Steven P. Tinling (Tinling et al., 2004), Otolaryngology Research Laboratory, University of California, Davis 95616.
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| Figure 9. Left Image. Image on the left is an electron micrograph of an inner hair cell (IHC) and supporting cells (S) from a Japanese macaque monkey. The lower left arrow indicates the region n at higher magnification in the image to the right. Bar = 3.0µm. Right Image. In the supporting cells next to the inner hair cell, the rough endoplasmic reticulum and microtubules are well-preserved. Bar = 0.5 µm. Reprinted with permission from Madden and Henson, 1997. From a paper on microwave accelerated decalcification by Madden and Henson, Department of Pathology and Laboratory Medicine, University of North Carolina, Chapel Hill, NC. |
六、參考文獻(xiàn)
- Galvez, J.J., Adamson, G., Sanders, M.A., Giberson, R.T., (2006) Microwave tissue processing techniques: their evolution and understanding. Microscopy and Analysis 20:23-24.
- Gerrity, R.G., Forbes, G.W., (2003) Microwave processing in diagnostic electron microscopy. Microsc. Today, 11(6):38-41.
- Gerrity RG, Forbes GW (2002) Microwave processing in diagnostic electron microscopy. Microsc Microanal 8(Suppl 2):152-153.
- Giberson, R.T., Austin, R.L., Charlesworth, J., Adamson, G., Herrera, G.A., (2003) Microwave and digital imaging technology reduce turnaround times for diagnostic electron microscopy. Ultrastruct. Pathol. 27:187-196.
- Wendt KD, Jensen CA, Tindall R, Katz ML, (2004) Comparison of conventional and microwave-assisted processing of mouse retinas for transmission electron microscopy. J Microsc 214:80-88
- Austin, RL (2002) The use of microwave technology in a clinical EM laboratory. Microsc Microanal 8(Suppl 2):154-155.
- Nordhausen RW, Barr BC, Hedrick RP (2002) Microwave-assisted rapid tissue processing for disease diagnosis in a veterinary diagnostic laboratory. Microsc Microanal 8(Suppl 2):150-151.
- Nordhausen RW, Barr BC (2001) Specimen preparation for this-section electron microscopy utilizing microwave-assisted rapid processing in a veterinary diagnostic laboratory. In Giberson RT, Demaree RSJr, eds. Microwave Techniques and Protocols, Totowa, NJ, Humana Press, 49-66.
- Giberson, RT., Elliott, DE (2001) Microwave-assisted formalin fixation of fresh tissue: A comparative study. In Giberson R.T., Demaree R.S.Jr., eds. Microwave Techniques and Protocols, Totowa, NJ, Humana Press, pp191-208.
- Galvez, JJ., Giberson, RT, Cardiff, RD (2006) The role of microwave radiation in reducing formaldehyde fixation times. The J. Histotechnol. 29:113-121.
- Galvez, JJ, Giberson, RT, Cardiff, RD (2004) Microwave mechanisms – the energy/heat dichotomy. Microsc. Today, 12(2):18-23.
- Munoz, TE, Giberson, RT, Demaree, R, Day JR (2004) Microwave-assisted immunostaining: a new approach yields fast and consistent results. J. Neurosci. Methods, 137:133-139.
- Sanders, M.A., Gartner, D.M. (2001) In vivo microwave-assisted labeling of Allium and Drosophila nuclei. In Giberson R.T., Demaree R.S.Jr., eds. Microwave Techniques and Protocols, Totowa, NJ, Humana Press, pp155-164.
- Madden, V.J., Henson, M.M. (1997) Rapid decalcification of temporal bones with preservation of ultrastructure. Hearing Research, 111;76-84.
- Tinling, S.P. Giberson, R.T., Kullar, R.S., (2004) Microwave exposure increases bone demineralization rate independent of temperature. J. Microsc., 215:230-235.
