Name: preparation of mouse retinal cryo-sections for immunohistochemistry
Uploaded: 2019-07-01T14:00:00
Description: Watch this Scientific Journal Video about Preparation of Mouse Retinal Cryo-sections for Immunohistochemistry at JoVE.com

这项工作得到了NIH(RO1-GMO97327)和大学研究基金会(UPenn)的资金支持。我们特别感谢斯韦特兰娜·萨维娜在开发免疫组织化学方案方面给予的帮助,戈登·鲁塞尔(宾夕法尼亚大学)和宾夕法尼亚大学Vet成像核心中心为显微镜和莱斯利·金博士提供了显微镜和莱斯利·金博士的辅助,帮助她阅读了这份手稿.

此处描述的所有方法都是严格按照国家卫生研究院《实验室动物护理和使用指南》中的建议进行的,并经宾夕法尼亚大学动物护理和使用机构委员会。
方法的所有工具和设备如图1所示,并列在材料表中。
1 .小鼠眼联、眼杯解剖和嵌入
<ol><li>用二氧化碳(CO2)对小鼠实施安乐死,然后斩首(产后小鼠P0 - P11)或宫颈脱位(?...

<ol>
	<li>Coons, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Labelled+antigens+and+antibodies.">Labelled antigens and antibodies.</a> Annual Review of Microbiology. 8, 333-352 (1954).</li><li>Coons, A. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorescent+antibodies+as+histochemical+tools.">Fluorescent antibodies as histochemical tools.</a> Federation Proceedings. 10 (2), 558-559 (1951).</li><li>Coons, A. H., Kaplan, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Localization+of+antigen+in+tissue+cells;+improvements+in+a+method+for+the+detection+of+antigen+by+means+of+fluorescent+antibody.">Localization of antigen in tissue cells; improvements in a method for the detection of antigen by means of fluorescent antibody.</a> Journal of Experimental Medicine. 91 (1), 1-13 (1950).</li><li>Chatterjee, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Artefacts+in+histopathology.">Artefacts in histopathology.</a> Journal of Oral Maxillofacial Pathology. 18 (Suppl 1), S111-S116 (2014).</li><li>Hartz, P. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Frozen+sections+from+Bouin-fixed+material+in+histopathology.">Frozen sections from Bouin-fixed material in histopathology.</a> Stain Technology. 20, 113 (1945).</li><li>Benerini Gatta, L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+alternative+fixatives+to+formalin+in+diagnostic+pathology.">Application of alternative fixatives to formalin in diagnostic pathology.</a> European Journal of Histochemistry. 56 (2), e12 (2012).</li><li>Tokuyasu, K. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+cryoultramicrotomy+to+immunocytochemistry.">Application of cryoultramicrotomy to immunocytochemistry.</a> Journal of Microscopy. 143 (Pt 2), 139-149 (1986).</li><li>Imai, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+properties+of+rhodopsin+and+rod+function.">Molecular properties of rhodopsin and rod function.</a> Journal of Biological Chemistry. 282 (9), 6677-6684 (2007).</li><li>Lee, J. W., Lim, M. Y., Park, Y. S., Park, S. J., Kim, I. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reexamination+of+Dopaminergic+Amacrine+Cells+in+the+Rabbit+Retina:+Confocal+Analysis+with+Double-+and+Triple-labeling+Immunohistochemistry.">Reexamination of Dopaminergic Amacrine Cells in the Rabbit Retina: Confocal Analysis with Double- and Triple-labeling Immunohistochemistry.</a> Experimental Neurobiology. 26 (6), 329-338 (2017).</li><li>Bringmann, A., Grosche, A., Pannicke, T., Reichenbach, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=GABA+and+Glutamate+Uptake+and+Metabolism+in+Retinal+Glial+(Muller)+Cells.">GABA and Glutamate Uptake and Metabolism in Retinal Glial (Muller) Cells.</a> Frontiers in Endocrinology (Lausanne). 4, 48 (2013).</li><li>Nakhai, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ptf1a+is+essential+for+the+differentiation+of+GABAergic+and+glycinergic+amacrine+cells+and+horizontal+cells+in+the+mouse+retina.">Ptf1a is essential for the differentiation of GABAergic and glycinergic amacrine cells and horizontal cells in the mouse retina.</a> Development. 134 (6), 1151-1160 (2007).</li><li>Applebury, M. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+murine+cone+photoreceptor:+a+single+cone+type+expresses+both+S+and+M+opsins+with+retinal+spatial+patterning.">The murine cone photoreceptor: a single cone type expresses both S and M opsins with retinal spatial patterning.</a> Neuron. 27 (3), 513-523 (2000).</li></ol>

由于小鼠眼睛的体积和形状小,视网膜易碎,小鼠视网膜解剖是一个微妙的过程。尽管进行高质量的解剖是一个实践问题,但拥有详细的协议,提供有效的方法和提示是获得视网膜部分和IHC的必要条件。除了此处介绍的协议外,还有几个提示允许一致的高质量视网膜部分,适用于可重复的 IHC。
在开始解剖之前,重要的是要舒适,放松,并设置房间照明,以在小鼠眼睛微解剖的最佳视力。为...

免疫组织化学(IHC)是一种强大的技术,用于在原位1、2、3定位组织的特定蛋白质和细胞结构。不适当的固定方法和复杂组织的次优切片可能会破坏组织结构,产生高背景染色或减少抗体-表皮相互作用,导致染色伪影,进而造成对IHC 数据4.由于脊椎动物视网膜是一种复杂且高度组织的神经器官,由相互连接的光受体、内神经细胞和神经细胞层组成,因此非常脆弱,在解剖和切片过程中很容易被破坏。从小鼠眼部解剖和定向到免疫染色的详细、标准化和经过验证的协议将有助于显著减少 IHC 伪影,从而提高结果的可靠性,并允许获得更准确的比较数据分析。
IHC的组织制备有许多方案,但是,并非所有方案都适合视网膜组织。强固定剂,如10%正式或布因的解决方案保留视网膜结构在解剖和切片5。不幸的是,由于表位6的化学修饰,强固定剂往往导致增强的背景荧光和表位掩蔽。另一方面,温和的固定剂,如4%的甲醛(PFA)可以减轻其中一些伪影,但需要细致的解剖和切片,以保持最佳的视网膜结构。PFA 迅速穿透组织,但交叉链接蛋白质非常缓慢,降低了表位掩蔽的风险。由于短时间的PFA孵育是一种相对温和的固定,组织往往需要快速冷冻,以保持抗原。在组织冻结期间避免冰晶形成是很重要的,因为它们会扭曲和损害细胞和组织的完整性7。
在这里,我们描述了用于小鼠眼后杯解剖、固定和冷冻保护的详细和标准化协议,这些电池杯可生成一致可靠的 IHC 数据。

<table border="0" cellpadding="0" cellspacing="0" style="width:1128px;" width="1129">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">6qt. Stainless Steel Beaker (185 x 218mm)</td>
			<td style="width:209px;">Gilson&#160;</td>
			<td style="width:236px;">#MA-48</td>
			<td style="width:365px;">For liquid nitrogen (Figure 1E)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Aluminum foil&#160;</td>
			<td style="width:209px;"></td>
			<td style="width:236px;"></td>
			<td style="width:365px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Cauterizer&#160;</td>
			<td style="width:209px;">Bovie</td>
			<td style="width:236px;">#AA01</td>
			<td style="width:365px;">To mark eye orientation (Figure 1A)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:317px;">Curved forceps, Dumont #5/45 tweezers, 45-degrees bent&#160;</td>
			<td style="width:209px;">Electron Microscopy Sciences</td>
			<td style="width:236px;">#72703-D</td>
			<td style="width:365px;">For mouse eye enucleation and maintenance (Figure 1A)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:317px;">Curved scissors, Vannas Spring Scissors - 2.5mm Cutting Edge&#160;</td>
			<td style="width:209px;">Fine Science Tools&#160;</td>
			<td style="width:236px;">#15002-08</td>
			<td style="width:365px;">To circumferentially cut the cornea (Figure 1B)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Dental wax-coated 35mm dissection dish</td>
			<td style="width:209px;"></td>
			<td style="width:236px;"></td>
			<td style="width:365px;">For eye cup dissection (Figure 1C)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:317px;">Dissecting microscope&#160;</td>
			<td style="width:209px;">Leica, Houston, USA&#160;</td>
			<td style="width:236px;">MZ12.5 High-performance stereomicroscope</td>
			<td style="width:365px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:317px;">Micro Knives - Plastic Handle&#160;</td>
			<td style="width:209px;">Fine Science Tools&#160;</td>
			<td style="width:236px;">#10315-12</td>
			<td style="width:365px;">To make a small incision at the burn mark (Figure 1A)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Pink dental wax&#160;</td>
			<td style="width:209px;">Electron Microscopy Sciences&#160;</td>
			<td style="width:236px;">#72660</td>
			<td style="width:365px;">For coating dissection dish</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Slide Rack and Coverplate</td>
			<td style="width:209px;">Ted Pella</td>
			<td style="width:236px;">#36107</td>
			<td style="width:365px;">For retinal IHC (Figure 1G)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Stainless Steel Beaker (89 x 114mm)</td>
			<td style="width:209px;">Gilson&#160;</td>
			<td style="width:236px;">#MA-40</td>
			<td style="width:365px;">For isopentane bath (Figure 1E)</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Styrofoam box</td>
			<td style="width:209px;"></td>
			<td style="width:236px;"></td>
			<td style="width:365px;">For insulated cooler</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:317px;">Thin forceps Dumont #5&#160;&#160;</td>
			<td style="width:209px;">Fine Science Tools&#160;</td>
			<td style="width:236px;">#11254-20</td>
			<td style="width:365px;">To remove the cornea and extract the lens (Figure 1B)</td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:317px;">Tissue-Tek cryomold&#160; (10mm x 10mm x 5mm)</td>
			<td style="width:209px;">Electron Miscroscopy Sciences&#160;</td>
			<td style="width:236px;">#62534-10</td>
			<td style="width:365px;">Mold for OCT embedding</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Buffers and Reagents</td>
			<td style="width:209px;">Company</td>
			<td style="width:236px;">Catalog Number</td>
			<td style="width:365px;">Comments</td>
		</tr>
		<tr height="58">
			<td height="58" style="height:58px;width:317px;">Blocking solution</td>
			<td style="width:209px;"></td>
			<td style="width:236px;"></td>
			<td style="width:365px;">2% Normal Horse Serum, 1.5% Cold Fish skin Gelatin (at 40-50% in H2O, cat #G7765), 5% bovine serum albumin (cat #AK1391-0100) in permeabilization buffer.&#160;&#160;</td>
		</tr>
		<tr height="77">
			<td height="77" style="height:77px;width:317px;">Gelvatol (anti-fading mounting media)</td>
			<td style="width:209px;"></td>
			<td style="width:236px;"></td>
			<td style="width:365px;">Under the fume hood, dissolve 0.337g DABCO (Sigma cat #D2522-25G) in 10mL Fluoromount G (Fisher cat #OB100-01).&#160; Adjust to pH 8-8.5 with 12N HCl (~ 5 drops).&#160; Store in amber drop bottle at 4&#176;C&#160; (8).&#160;</td>
		</tr>
		<tr height="58">
			<td height="58" style="height:58px;width:317px;">Hoechst 33342 1000x Stock</td>
			<td style="width:209px;">Thermo Scientific</td>
			<td style="width:236px;">#62249</td>
			<td style="width:365px;">1 mg/ml in PBS (1000x). Aliquot and store in dark, at 4&#176;C for up to 1 year or at -20&#176;C for long term storage. Prepare fresh&#160; at 1 ug/mL (1x)&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Isopentane, 99%&#160;</td>
			<td style="width:209px;">GFS Chemicals</td>
			<td style="width:236px;">#2961</td>
			<td style="width:365px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Liquid Nitrogen</td>
			<td style="width:209px;"></td>
			<td style="width:236px;"></td>
			<td style="width:365px;"></td>
		</tr>
		<tr height="38">
			<td height="38" style="height:38px;width:317px;">Paraformaldehyde (PFA) in PBS</td>
			<td style="width:209px;">Electron Microscopy Sciences&#160;</td>
			<td style="width:236px;">#15710</td>
			<td style="width:365px;">Prepared fresh 4% PFA from 16% PFA stock. Store the remaining 16% PFA at 4&#176;C in the dark.&#160;</td>
		</tr>
		<tr height="38">
			<td height="38" style="height:38px;width:317px;">Permeabilization solution</td>
			<td style="width:209px;"></td>
			<td style="width:236px;"></td>
			<td style="width:365px;">PBS + 0.25% Triton-X (AMRESCO cat #0694-1L) + 0.05% NaN3.&#160; Prepare fresh.</td>
		</tr>
		<tr height="50">
			<td height="50" style="height:50px;width:317px;">Phosphate-buffered saline (PBS)</td>
			<td style="width:209px;">Bioworld&#160;</td>
			<td style="width:236px;">#41620015-20</td>
			<td style="width:365px;">0.02 g/L KCl, 0.02 g/L KH2PO4, 0.8 g/L NaCl, 0.216 g/L Na2HPO4 , pH 7.4.&#160; Prepared from 10x stock&#160;</td>
		</tr>
		<tr height="58">
			<td height="58" style="height:58px;width:317px;">Sucrose solutions</td>
			<td style="width:209px;">Fisher Scientific&#160;&#160;</td>
			<td style="width:236px;">#S-5-500</td>
			<td style="width:365px;">Dissolve the appropriate amount of sucrose in&#160; 37&#176;C PBS (takes ~15 min to dissolve). May be stored for a few days at 4&#176;C.&#160;</td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Tissue-Tek OCT-compound&#160;</td>
			<td style="width:209px;">Sakura</td>
			<td style="width:236px;">&#160;#4583</td>
			<td style="width:365px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:317px;">Antibodies</td>
			<td style="width:209px;">Company</td>
			<td style="width:236px;">Catalog Number</td>
			<td style="width:365px;">Comments</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:317px;">anti-calbindin</td>
			<td>Sigma-Aldrich</td>
			<td>C8666</td>
			<td>To label horinzotal cells&#160;</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:317px;">anti-GAD65</td>
			<td>Chemicon</td>
			<td>AB5082</td>
			<td>To label GABAergic amacrine cells</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:317px;">anti-GS</td>
			<td>BD Transduction</td>
			<td>610517</td>
			<td>To label M&#252;ller cells</td>
		</tr>
		<tr height="19">
			<td height="19" style="height:19px;width:317px;">anti-rhodopsin</td>
			<td>Millipore</td>
			<td>MAB5316</td>
			<td>To label rods</td>
		</tr>
	</tbody>
</table>

preparation of mouse retinal cryo-sections for immunohistochemistry

为免疫组织化学 (IHC) 制备高质量的小鼠眼部部分对于评估视网膜结构和功能以及确定视网膜疾病背后的机制至关重要。在整个组织制备过程中保持结构完整性对于获得可重复的视网膜 IHC 数据至关重要,但由于视网膜细胞结构的脆弱性和复杂性,这些数据可能具有挑战性。强固定剂,如10%形式蛋白或布恩的溶液,以最佳方式保持视网膜结构,它们通常通过增强背景荧光和/或减少抗体-表位相互作用(称为表位掩蔽的过程)来阻碍IHC分析。另一方面,较温和的固定剂,如4%的甲醛,减少背景荧光和表位遮蔽,必须采用细致的解剖技术来保持视网膜结构。在本文中,我们将介绍一种为 IHC 制备小鼠眼后杯的综合方法,该方法足以保留大多数抗体-表皮相互作用,而不会损失视网膜结构完整性。我们包括具有代表性的IHC与抗体的各种视网膜细胞类型标记,以说明组织保存和方向在最佳和次优条件下。我们的目标是通过提供从眼后杯解剖到IHC的完整方案来优化视网膜的IHC研究。

为了说明这些协议如何,确保IHC的最佳视网膜保存,我们用对红斑素(光受体标记物)8、谷氨酸酸脱碳酶65(Gad65,一种丙烯细胞)的抗体对P28 WT小鼠(C57BL/6N)的视网膜部分进行了调查标记)9, 谷氨酰胺合成酶 (GS, Müller 细胞标记)10,和钙化素 (水平细胞标记)11 (图 4)。IHC 表明我们的协议始终能...

Watch this Scientific Journal Video about Preparation of Mouse Retinal Cryo-sections for Immunohistochemistry at JoVE.com

Preparation of Mouse Retinal Cryo-sections for Immunohistochemistry

本报告介绍了制备冷冻小鼠视网膜部分用于免疫组织化学(IHC)的综合方法。 所述方法包括眼部后杯的解剖、甲醛固定、嵌入最佳切割温度(OCT)介质和组织定向、切片和免疫染色。

小鼠视网膜冷冻节的制备,用于免疫性植物化学

Preparation of high-quality mouse eye sections for immunohistochemistry (IHC) is critical for assessing the retinal structure and function and for ...

由于眼睛的大小和形状以及视网膜部分的脆弱性，小鼠视网膜解剖是一个非常微妙的过程。实践势在必行，有一个详细的协议，提供有效的方法和提示是高品质的视网膜解剖和部分的关键。我们提供提示和建议，以帮助研究人员避免常见的陷阱，并获得一些高品质的视网膜部分适合免疫组织化学。使用尾烧灼伤器轻轻触摸角膜，用一瞬间轻轻触摸被安乐死小鼠的眼睛的时间部分，以稍微灼伤角膜，避免打洞。紧接着，使用弯曲的杜蒙545钳子，以羡慕老鼠的眼睛。将眼睛放在1.5毫升的冷冻管中，用1毫升4%PFA和PBS，在冰上孵育15分钟。然后，使用弯曲的Dumont 545钳子将固定的眼睛转移到一个经过修饰的35毫米解剖盘上，里面装满了PBS，并放在解剖显微镜下。使用微刀在垂直于角膜边缘边缘的烧伤标记上做一个小切口。将弯曲的剪刀插入切口，并在边缘后对角膜进行圆周切割。然后，使用薄的Dumont 5钳子，取出角膜，提取透镜，并小心翼翼地将其从眼睛的后部提起。玻璃体会与透镜一起出来，视网膜将可见为覆盖后眼杯内侧的白色表面。在室温下将隔离眼罩在一毫升 PBS 中清洗两次，10 分钟。要冷冻保护眼罩，在4摄氏度下将眼杯在15%蔗糖和PBS中平衡，直到它们下沉。然后将眼罩转移到30%蔗糖和PBS两到三个小时，直到它们下沉。将眼罩放在 10 百万分之一或 10 毫米低温模具中，用于嵌入。在解剖显微镜下，通过旋转眼睛，将低温模具中的眼睛沿着侧腹轴定向，直到烧伤标记位于顶部。视神经是左边的神经，模子的切口部分朝右。要捕捉冷冻视网膜组织，将低温模具浸入含有异丙烷的金属烧杯中至少五分钟，然后将烧杯放在液氮中，达到烧杯高度的三分之一。取出冷冻块，用铝箔包裹，储存在零下80摄氏度。使用笔或夏皮标记冻结块以记录其方向。将低温恒温调整到零下 20 至零下 25 摄氏度之间后，将含有嵌入眼罩的模具放在里面，让其与低温温度均衡一小时。将块安装到低温恒温器中，并带后腹方向，开始切割 10 微米厚的串行截面。小心地将视网膜部分放在带注释和编号的显微镜幻灯片上，然后存放在零下 20 摄氏度或零下 80 摄氏度的幻灯片盒中，直到准备好进行免疫破坏。免疫hito化学与抗体的罗多普辛，光受体标记，谷氨酸脱卡盒酶65，和阿马克林细胞标记，谷氨酰胺合成和穆勒细胞标记，和卡宾丁，在视网膜部分执行水平细胞标记。结果表明，该协议产生高质量和保存完好的视网膜部分，不同于从不同协议获得的劣质视网膜部分。富含罗多普辛丰富的内层和外段保持垂直和完好，很少或没有分离，从覆盖视网膜色素上皮。内部肿瘤，如Gad65正阿马克林细胞，保持适当的分层内核层和内层。此外，穆勒细胞在从结节细胞层到外核层的细胞质过程中，与索马正确对齐，保存完好。此外，在内部核层和外层边界上可检测到定义良好的水平细胞。标记眼睛的顶点区域并在嵌入过程中保持方向非常重要。始终使用适当的个人防护设备在引擎盖下准备半甲醛。

Research

JoVE Journal

Neuroscience

Preparation of Mouse Brain Tissue for Immunoelectron Microscopy

制备小鼠脑组织中的免疫电镜

Transmission electron microscopy (TEM) is extremely useful for visualizing microglial, oligodendrocytic, astrocytic, and neuronal subcellular compartments ...

科研

神经科学

Patch Clamp Recordings from Mouse Retinal Neurons in a Dark-adapted Slice Preparation

从一个暗适应切片制备小鼠视网膜神经细胞的膜片钳录音

Our visual experience is initiated when the visual pigment in our retinal photoreceptors absorbs photons of light energy and initiates a cascade of ...

An Isolated Retinal Preparation to Record Light Response from Genetically Labeled Retinal Ganglion Cells

一个孤立的视网膜制备，记录从基因标记的视网膜神经节细胞的光反应

The first steps in vertebrate vision take place when light stimulates the rod and cone photoreceptors of the retina 1.  This information is then ...

Dissection of a Mouse Eye for a Whole Mount of the Retinal Pigment Epithelium

解剖鼠标眼的视网膜色素上皮的全山

The retinal pigment epithelium (RPE) lies at the back of the mammalian eye, just under the neural retina, which contains the photoreceptors (rods and ...

Preparation of Non-human Primate Brain Tissue for Pre-embedding Immunohistochemistry and Electron Microscopy

非人类灵长类动物脑组织的制备预嵌入免疫组化和电子显微镜

Despite all the technological advances at the light microscopy&#160;level, electron microscopy remains the only tool in neuroscience to examine and ...

Optical Coherence Tomography: Imaging Mouse Retinal Ganglion Cells In Vivo

Optical Coherence Tomography: Imaging Mouse Retinal Ganglion Cells In Vivo

光学相干断层扫描: 成像小鼠视网膜神经节细胞在体内

Structural changes in the retina are common manifestations of ophthalmic diseases. Optical coherence tomography (OCT) enables their identification in ...

Optimization of the Retinal Vein Occlusion Mouse Model to Limit Variability

Mouse models of retinal vein occlusion (RVO) are often used in ophthalmology to study hypoxic-ischemic injury in the neural retina. In this report, a ...

Mouse Retina Isolation and Methanol Treatment for Studying Retinal Ganglion Cells

Methanol-Based Whole-Mount Preparation for the Investigation of Retinal Ganglion Cells

Methanol-Based Whole-Mount Preparation for the Investigation of Retinal Ganglion Cells (Video) | JoVE 

基于甲醇的全支架制备用于视网膜神经节细胞的研究

Retinal ganglion cells (RGCs), which are the projection neurons of the retina, propagate external visual information to the brain. Pathological changes in ...

Detection of Abnormal Prion Protein by Immunohistochemistry

免疫组化检测异常朊蛋白

Abnormal prion proteins (PrPSc) are the disease-associated isoform of cellular prion protein and diagnostic markers of transmissible spongiform ...

Mouse Retina OCT Fibergram Alignment with Confocal Images

Alignment of Visible-Light Optical Coherence Tomography Fibergrams with Confocal Images of the Same Mouse Retina

Advancements in In Vivo and Ex Vivo Retinal Imaging for Improved Glaucoma Diagnosis and Treatment (Video) | JoVE

可见光光学相干断层扫描光纤图与同一小鼠视网膜的共聚焦图像的对齐

In recent years, in vivo retinal imaging, which provides non-invasive, real-time, and longitudinal information about biological systems and processes, has ...