VBM 得到 NIH 的资助 [K08EY020530, R01EY024665, R01EY025225, R01EY024698 和 R21AG050437], 桃乐丝杜克慈善基金会补助金: 2013103, 并研究, 以防止失明 (填料), 纽约, 纽约州。MT 和 T32GM007337 得到 NIH 的资助。

这项研究是由爱荷华大学和 #39 的机构审查委员会批准的, 并遵循《赫尔辛基宣言》中所规定的原则. 
 1. 中心和黄斑活检穿孔 <ol> <li> 打开和蝶形人眼, 如前一出版物所述, 它由4独立的组织瓣组成. 5 </li> <li> 开始时, butterflied 人眼放置在培养皿中, 将一个 4 mm 的穿孔活检工具放在凹窝上, 向下按压, 然后轻轻滚动, 直到在凹腹周围进行切口. </li>
	<li> 接下来, 在黄斑的拱廊内, 通过居中并按下一个 8 mm 的皮肤穿孔活检工具, 在黑点周围产生切口, 施加柔和的压力和滚动。这将产生第二个外环的组织周围的第一. </li>
</ol> 2。外周视网膜活检穿孔 <ol> <li> 使用 4 mm 的皮肤穿孔活检工具在外围视网膜上进行一系列击打, 就在拱廊外。在这里, 为每个象限襟翼做两拳. 
	注: 在所有的拳打后, 眼睛将有两个同心拳在中心, 代表中央凹和黄斑, 两个拳在底部的每瓣-总计8拳在周边视网膜. </li>
</ol> 3。凹和黄斑活检收集 <ol> <li> 当组织现在准备好收集, 收集所有组织活检在分开的离心管和冷冻使用液氮为下游处理。将所有样品存储在-80 和 #176; C 直到使用. </li>
	<li> 使用弯曲的 0.12 Colibri 钳抓住视网膜凹的半透明组织的边缘。从底层视网膜色素上皮中收集、提升和分离中心组织. </li>
	<li> 同样, 使用 0.12 Colibri 钳来抓住半透明的黄斑组织的外环。如果组织仍然连接到基础或相邻组织, 使用一副保罗·韦斯科特剪刀仔细修剪边缘, 捕获只是视网膜成分和解剖的任何视神经组织包括在冲床. </li>
</ol> 4。周边视网膜采集 <ol> <li> 使用弯曲的 0.12 Colibri 钳将周围的视网膜组织瓣轻轻分开, 从底层的 RPE-脉络膜, 并在单个离心管中放置. </li>
	<li> 在残留的玻璃体凝胶的情况下, 使用镊子解除和分离的凝胶尽可能多之前收集的视网膜组织。一旦视网膜被移除, 色素 RPE-脉络膜仍在下方. </li>
</ol> 5。中心和黄斑视网膜色素上皮集合 <ol> <li> 使用 0.12 Colibri 钳来抓住暗 RPE-脉络膜组织的边缘, 位于所移除凹区的下方。仔细分离这个组织从巩膜和收集. </li>
	<li> 使用同样的钳子, 抓住边缘的黑暗 RPE-脉络膜组织的基础上清除黄斑区。仔细地把这个组织从巩膜上分离出来, 抓住环周围不同点的边缘, 轻轻地拉。最终, RPE-脉络膜组织的环会脱落, 并可能被收集. 
	注意: 相反手上的二次钳在去除视网膜色素上皮组织的过程中可以起到帮助作用。像视网膜组织, Wescott 剪刀可以用来帮助删除任何组织, 没有完全切割使用冲床工具. </li>
</ol> 6。外周视网膜 rpe-脉络膜集合 <ol> <li> 使用 0.12 Colibri 钳将 RPE-脉络膜剥离到8外围冲头区域. </li>
	<li> 如前所述, 将 RPE-脉络膜组织置于离心管中, 并使用液氮进行下游加工。将所有样品保持在-80 和 #176; C 直到使用. </li>
</ol> 7。剪刀解剖 注意: 如果穿孔活检刀片是钝的或穿孔活检工具不够用力, 可能不会有一个 clean-cut 周围的组织. 
<ol> <li> 在这些情况下, 尽可能地将组织拉离, 然后使用保罗·韦斯科特剪刀修剪和分离. </li> </ol>

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未声明任何利益冲突。

组织收集后, 样品处理和处理是至关重要的考虑因素14。液氮的保存优于化学固定, 因为后者可能导致蛋白质结构的破坏, 这可能会扭曲下游分析。此外, 液氮保存是首选的方法, 不涉及冻结样品。值得注意的是, 与0° c15相比, Ferrer et al.在4° c 或室温下保存的脑样品的蛋白质水平有显著差异。此外, 重要的是要小心, 当处理样品, 由于特定的化学和物理治疗前?...

视网膜、RPE 和脉络膜是复杂的组织, 在蛋白质表达、生理功能和病理敏感性方面表现出重要的区域差异1,2。例如, 与年龄相关的黄斑变性 (AMD)、视网膜色素变性和中央浆液性视网膜病变等疾病都表现出在凹、黄斑或视网膜周围的特征定位1,3, 4,6。在这里, 我们提出了一个方法来演示如何独立取样的视网膜区域。该方法的总体目标是提供一个可靠的指南, 收集的组织样本从中心, 黄斑, 周边地区的人视网膜和 RPE-脉络膜的蛋白质组分析。这一技术的发展和应用的基本原理是, 通过对这些特定的视网膜区域的蛋白质组分析, 可以获得重要的分子洞察力, 这些区域的生理和病理功能。
这种方法有望揭示相关区域疾病敏感性的蛋白质组基础, 并有助于确定新的具体治疗目标。事实上, 蛋白质组学对玻璃体及其与视网膜相互作用的研究为健康和患病组织的分子组成和功能提供了重要的洞察力5,7,8,9,10,11,12,13. 然而, 对不同的视网膜区域缺乏明确的比较蛋白质组分析。该技术将有助于支持这些急需的研究, 提供比其他方法的优势, 通过展示一个可靠的和可再生的组织收集方法。更重要的是, 该方法是非常容易获得的, 利用标准和现成的组织穿孔活检工具。我们的技术强调适当的收集和储存组织的蛋白质组处理, 使重要的考虑蛋白的稳定性和降解。因此, 这种方法是最适合的研究者考虑下游分子分析蛋白质的因素。

<table><tbody><tr><td>4-mm skin punch biopsy tool</td><td>Miltex</td><td>REF 33-34</td><td></td></tr><tr><td>8-mm skin punch biopsy tool</td><td>Miltex</td><td>REF 33-37</td><td></td></tr><tr><td>0.12 Colibri Forceps</td><td>Stephens Instruments</td><td>S5-1145</td><td></td></tr><tr><td>Wescott Scissors</td><td>Sklar Surgical Instruments</td><td>64-3146</td><td></td></tr><tr><td>Microfuge tubes</td><td>Eppendorf</td><td>#022364111</td><td>1.5 mL</td></tr><tr><td>Liquid Nitrogen</td><td>Praxair, Inc.</td><td>7727-37-9 [R]</td><td></td></tr></tbody></table>

dissection of human retina and rpe-choroid for proteomic analysis

人视网膜由感觉 neuroretina 和底层的视网膜色素上皮 (RPE) 组成, 它与血管脉络膜层紧密地复合。视网膜的不同区域是解剖学上和分子上的区别, 促进独特的功能和证明差异易感性的疾病。蛋白质组分析的每个区域和层可以提供重要的洞察力的分子过程中的许多疾病, 包括年龄相关的黄斑变性 (AMD), 糖尿病和青光眼。然而, 在进行定量蛋白质组分析之前, 视网膜区域和层的分离是必不可少的。在这里, 我们描述了解剖和收集的中心, 黄斑, 和周边视网膜区域和底层 RPE-脉络膜复合体的方法, 涉及区域穿孔活检和手工清除组织层从人眼。一维 SDS 页以及下游蛋白质组学分析, 如液相色谱-串联质谱 (LC-ms), 可用于识别每个解剖视网膜层的蛋白质, 揭示视网膜疾病的分子标志物。

视网膜和 RPE-脉络膜组织可以以各种方式进行处理, 以适应个人的调查。收集后, 研究员将拥有视网膜和 RPE-脉络膜组织从中心地区, 外黄斑和周边视网膜 (图 1) 的样本。具体来说, 中心区域冲床将包括凹, parafovea, 和少量的相邻 perifovea。黄斑冲头包括 perifoveal 区的其余部分以及邻近近周边区域的少量。最后, 对 mid-peripheral 和远外围区域进行了外围冲压?...

Watch this Scientific Journal Video about Dissection of Human Retina and RPE-Choroid for Proteomic Analysis at JoVE.com

Dissection of Human Retina and RPE-Choroid for Proteomic Analysis

人类视网膜由功能和分子的不同区域组成, 包括黄斑、黄斑和外周视网膜。在这里, 我们描述了一个方法使用穿孔活检和手工清除组织层从人眼解剖和收集这些不同的视网膜区域的蛋白质组分析。

人视网膜解剖与 RPE-脉络膜的蛋白质组分析

The human retina is composed of the sensory neuroretina and the underlying retinal pigmented epithelium (RPE), which is firmly complexed to the vascular ...

dissection-of-human-retina-and-rpe-choroid-for-proteomic-analysis

Universidad de Cartagena UDEC

Research

JoVE Journal

Biochemistry

10.8K 次观看.  Columbia University. 人类视网膜由功能和分子的不同区域组成, 包括黄斑、黄斑和外周视网膜。在这里, 我们描述了一个方法使用穿孔活检和手工清除组织层从人眼解剖和收集这些不同的视网膜区域的蛋白质组分析。

视频: Dissection of Human Retina and RPE-Choroid for Proteomic Analysis

Experimental Models for Study of Retinal Pigment Epithelial Physiology and Pathophysiology

We have developed a cell culture procedure that can produce large quantities of confluent monolayers of primary human fetal retinal pigment epithelium (hfRPE) cultures with morphological, physiological and genetic characteristics of native human RPE. These hfRPE cell cultures exhibit heavy pigmentation, and electron microscopy show extensive apical membrane microvilli. The junctional complexes were identified with immunofluorescence labeling of various tight junction proteins. Epithelial polarity and function of these easily reproducible primary cultures closely resemble previously studied mammalian models of native RPE, including human. These results were extended by the development of therapeutic interventions in several animal models of human eye disease. We have focused on strategies for the removal of abnormal fluid accumulation in the retina or subretinal space. The extracellular subretinal space separates the photoreceptor outer segments and the apical membrane of the RPE and is critical for maintenance of retinal attachments and a whole host of RPE/retina interactions.

We provide a reproducible method for culturing confluent monolayers of human fetal retinal pigment epithelial cells (hfRPE) cells that exhibit morphology, physiology, polarity, and protein and gene expression patterns of adult native tissue. This work has been extended to an animal model of several eye diseases.

视网膜色素上皮的生理与病理生理学研究的实验模型

We have developed a cell culture procedure that can produce large quantities of confluent monolayers of primary human fetal retinal pigment epithelium ...

科研

神经科学

Dissection of Human Vitreous Body Elements for Proteomic Analysis

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. 1,2 Abnormal interactions between vitreous substructures and the retina underlie several vitreoretinal diseases, including retinal tear and detachment, macular pucker, macular hole, age-related macular degeneration, vitreomacular traction, proliferative vitreoretinopathy, proliferative diabetic retinopathy, and inherited vitreoretinopathies. 1,2 The molecular composition of the vitreous substructures is not known. Since the vitreous body is transparent with limited surgical access, it has been difficult to study its substructures at the molecular level. We developed a method to separate and preserve these tissues for proteomic and biochemical analysis. The dissection technique in this experimental video shows how to isolate vitreous base, anterior hyaloid, vitreous core, and vitreous cortex from postmortem human eyes. One-dimensional SDS-PAGE analyses of each vitreous component showed that our dissection technique resulted in four unique protein profiles corresponding to each substructure of the human vitreous body. Identification of differentially compartmentalized proteins will reveal candidate molecules underlying various vitreoretinal diseases.

This video shows an effective technique for differentiating and dissecting the various semi-transparent structures of the human vitreous body in post mortem eyes.

人类蛋白质组分析玻璃体元素剖析

The vitreous is an optically clear, collagenous extracellular matrix that fills the inside of the eye and overlies the retina. 1,2 Abnormal interactions ...

医学

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

The retinal pigment epithelium (RPE) lies at the back of the mammalian eye, just under the neural retina, which contains the photoreceptors (rods and cones). The RPE is a monolayer of pigmented cuboidal cells and associates closely with the neural retina just above it. This association makes the RPE of great interest to researchers studying retinal diseases. The RPE is also the site of an in vivo assay of homology-directed DNA repair, the pun assay. The mouse eye is particularly difficult to dissect due to its small size (about 3.5mm in diameter) and its spherical shape. This article demonstrates in detail a procedure for dissection of the eye resulting in a whole mount of the RPE. In this procedure, we show how to work with, rather than against, the spherical structure of the eye. Briefly, the connective tissue, muscle, and optic nerve are removed from the back of the eye. Then, the cornea and lens are removed. Next, strategic cuts are made that result in significant flattening of the remaining tissue. Finally, the neural retina is gently lifted off, revealing an intact RPE, which is still attached to the underlying choroid and sclera. This whole mount can be used to perform the pun assay or for immunohistochemistry or immunofluorescent assessment of the RPE tissue.

A formal demonstration of the dissection of a mouse eye, resulting in 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 ...

Single-cell Profiling of Developing and Mature Retinal Neurons

Highly specialized, but exceedingly small populations of cells play important roles in many tissues. The identification of cell-type specific markers and gene expression programs for extremely rare cell subsets has been a challenge using standard whole-tissue approaches. Gene expression profiling of individual cells allows for unprecedented access to cell types that comprise only a small percentage of the total tissue1-7. In addition, this technique can be used to examine the gene expression programs that are transiently expressed in small numbers of cells during dynamic developmental transitions8.
This issue of cellular diversity arises repeatedly in the central nervous system (CNS) where neuronal connections can occur between quite diverse cells9. The exact number of distinct cell types is not precisely known, but it has been estimated that there may be as many as 1000 different types in the cortex itself10. The function(s) of complex neural circuits may rely on some of the rare neuronal types and the genes they express. By identifying new markers and helping to molecularly classify different neurons, the single-cell approach is particularly useful in the analysis of cell types in the nervous system. It may also help to elucidate mechanisms of neural development by identifying differentially expressed genes and gene pathways during early stages of neuronal progenitor development.
As a simple, easily accessed tissue with considerable neuronal diversity, the vertebrate retina is an excellent model system for studying the processes of cellular development, neuronal differentiation and neuronal diversification. However, as in other parts of the CNS, this cellular diversity can present a problem for determining the genetic pathways that drive retinal progenitors to adopt a specific cell fate, especially given that rod photoreceptors make up the majority of the total retinal cell population11. Here we report a method for the identification of the transcripts expressed in single retinal cells (Figure 1). The single-cell profiling technique allows for the assessment of the amount of heterogeneity present within different cellular populations of the retina2,4,5,12. In addition, this method has revealed a host of new candidate genes that may play role(s) in the cell fate decision-making processes that occur in subsets of retinal progenitor cells8. With some simple adjustments to the protocol, this technique can be utilized for many different tissues and cell types.

A method for the isolation of single retinal cells and subsequent amplification of their cDNAs is described. Single-cell transcriptomics reveals the degree of cellular heterogeneity present in a tissue and uncovers new marker genes for rare cell populations. The accompanying protocol can be adjusted to suit many different cell types.

视网膜神经细胞发育和成熟的单细胞的剖析

Highly specialized, but exceedingly small populations of cells play important roles in many tissues. The identification of cell-type specific markers and ...

The Gateway to the Brain: Dissecting the Primate Eye

The visual system in humans is considered the gateway to the world and plays a principal role in the plethora of sensory, perceptual and cognitive processes. It is therefore not surprising that quality of vision is tied to quality of life . Despite widespread clinical and basic research surrounding the causes of visual disorders, many forms of visual impairments, such as retinitis pigmentosa and macular degeneration, lack effective treatments. Non-human primates have the closest general features of eye development to that of humans. Not only do they have a similar vascular anatomy, but amongst other mammals, primates have the unique characteristic of having a region in the temporal retina specialized for high visual acuity, the fovea1. Here we describe a general technique for dissecting the primate retina to provide tissue for retinal histology, immunohistochemistry, laser capture microdissection, as well as light and electron microscopy. With the extended use of the non-human primate as a translational model, our hope is that improved understanding of the retina will provide insights into effective approaches towards attenuating or reversing the negative impact of visual disorders on the quality of life of affected individuals.

The non-human primate is an important translational species for our understanding of development and aging. The anatomical organization of the primate retina may provide important insights into normal and pathological conditions in humans.

到大脑的门户：剖析灵长类动物的眼睛

The visual system in humans is considered the  gateway  to the world and plays a principal role in the plethora of sensory, perceptual and cognitive ...

生物学

A Protocol to Evaluate and Quantify Retinal Pigmented Epithelium Pathologies in Mouse Models of Age-Related Macular Degeneration

Age-related macular degeneration (AMD) is a debilitating retinal disorder in aging populations. It is widely believed that dysfunction of the retinal pigmented epithelium (RPE) is a key pathobiological event in AMD. To understand the mechanisms that lead to RPE dysfunction, mouse models can be utilized by researchers. It has been established by previous studies that mice can develop RPE pathologies, some of which are observed in the eyes of individuals diagnosed with AMD. Here, we describe a phenotyping protocol to assess RPE pathologies in mice. This protocol includes the preparation and evaluation of retinal cross-sections using light microscopy and transmission electron microscopy, as well as that of RPE flat mounts by confocal microscopy. We detail the common types of murine RPE pathologies observed by these techniques and ways to quantify them through unbiased methods for statistical testing. As proof of concept, we use this RPE phenotyping protocol to quantify the RPE pathologies observed in mice overexpressing transmembrane protein 135 (Tmem135) and aged wild-type C57BL/6J mice. The main goal of this protocol is to present standard RPE phenotyping methods with unbiased quantitative assessments for scientists using mouse models of AMD.

Mouse models can be useful tools for investigating the biology of the retinal pigmented epithelium (RPE). It has been established that mice can develop an array of RPE pathologies. Here, we describe a phenotyping protocol to elucidate and quantify RPE pathologies in mice using light, transmission electron, and confocal microscopy.

评估和量化年龄相关性黄斑变性小鼠模型中视网膜色素上皮病理的协议

Age-related macular degeneration (AMD) is a debilitating retinal disorder in aging populations. It is widely believed that dysfunction of the retinal ...

遗传学

Microdissection of Zebrafish Embryonic Eye Tissues

Zebrafish is a popular animal model for research on eye development because of its rapid ex utero development and good fecundity. By 3 days post fertilization (dpf), the larvae will show the first visual response. Many genes have been identified to control a proper eye development, but we are far from a complete understanding of the underlying genetic architecture. Whole genome gene expression profiling is a useful tool to elucidate genetic regulatory network for eye development. However, the small size of the embryonic eye in zebrafish makes it challenging to obtain intact and pure eye tissues for expression analysis. For example, the anterior-posterior length of the eye between day 2 and 3 is only approximately 200-300 &mu;m, while the diameter of the lens is less 100 &mu;m. Also, the retinal pigment epithelium (RPE) underlying the retina is just a single-layer epithelium. While gene expression profiles can be obtained from the whole embryo, they do not accurately represent the expression of these tissues. Therefore pure tissue must be obtained for a successful gene expression profiling of eye development. To address this issue, we have developed an approach to microdissect intact retina and retina with RPE attached from 1-3 dpf, which cover major stages of eye morphogenesis. All procedures can be done with fine forceps and general laboratory supplies under standard stereomicroscopes. For retinal dissection, the single-layer RPE is removed and peeled off by brushing action and the preferential adherence of the RPE remnants to the surface of the culture plate for dissection. For RPE-attached retinal dissection, the adherence of RPE to the dissection plate is removed before the dissection so that the RPE can be completely preserved with the retina. A careful lifting action of this tissue can efficiently separate the presumptive choroid and sclera. The lens can be removed in both cases by a chemically etched tungsten needle. In short, our approach can obtain intact eye tissues and has been successfully utilized to study tissue-specific expression profiles of zebrafish retina1, 2 and retinal pigment epithelium3.

This article describes an approach to microdissect zebrafish retinas with and without retinal pigment epithelium attached, from one to three days postfertilization embryos.

斑马鱼胚胎眼组织显微切割的

Zebrafish is a popular animal model for research on eye development because of its rapid ex utero development and good fecundity. By 3 days post ...

Evisceration of Mouse Vitreous and Retina for Proteomic Analyses

While the mouse retina has emerged as an important genetic model for inherited retinal disease, the mouse vitreous remains to be explored. The vitreous is a highly aqueous extracellular matrix overlying the retina where intraocular as well as extraocular proteins accumulate during disease.1-3 Abnormal interactions between vitreous and retina underlie several diseases such as retinal detachment, proliferative diabetic retinopathy, uveitis, and proliferative vitreoretinopathy.1,4 The relative mouse vitreous volume is significantly smaller than the human vitreous (Figure 1), since the mouse lens occupies nearly 75% of its eye.5 This has made biochemical studies of mouse vitreous challenging. In this video article, we present a technique to dissect and isolate the mouse vitreous from the retina, which will allow use of transgenic mouse models to more clearly define the role of this extracellular matrix in the development of vitreoretinal diseases.

The dissection technique illustrates evisceration of the vitreous, retina, and lens from the mouse eye, separation by centrifugation, and characterization with protein assays.

眼内容剜除小鼠玻璃体和视网膜蛋白质组分析

While the mouse retina has emerged as an important genetic model for inherited retinal disease, the mouse vitreous remains to be explored. The vitreous is ...

Microsurgical Dissection and Tissue Clearing for High Resolution Intact Whole Retina and Vitreous Imaging

Neuronal and vascular structures of the retina in physiologic and pathologic conditions can be better visualized and characterized by using intact whole retina imaging techniques compared to conventional retinal flat mount preparations and sections. However, immunofluorescent imaging of intact whole retina is hindered by the opaque coatings of the eyeball, i.e., sclera, choroid, and retinal pigment epithelium (RPE) and the light scattering properties of retinal layers that prevent full thickness high resolution optical imaging. Chemical bleaching of the pigmented layers and tissue clearing protocols have been described to address these obstacles; however, currently described methods are not suitable for imaging endogenous fluorescent molecules such as green fluorescent protein (GFP) in intact whole retina. Other approaches bypassed this limitation by surgical removal of pigmented layers and the anterior segment of the eyeball allowing intact eye imaging, though the peripheral retina and hyaloid structures were disrupted. Presented here is an intact whole retina and vitreous immunofluorescent imaging protocol that combines surgical dissection of the sclera/choroid/retina pigment epithelium (RPE) layers with a modified tissue clearing method and light sheet fluorescent microscopy (LSFM). The new approach offers an unprecedented view of&#160;unperturbed vascular and neuronal elements of the retina as well as the vitreous and hyaloid vascular system in pathologic conditions.

Presented here is a protocol for intact whole retina imaging in which the outer opaque/pigmented layers of the eyeball are surgically removed, and optical clearing is applied to render retina transparent enabling the visualization of the peripheral retina and hyaloid vasculature in intact retina using light sheet fluorescent microscopy.

Neuronal and vascular structures of the retina in physiologic and pathologic conditions can be better visualized and characterized by using intact whole ...

Two Peeling Methods for the Isolation of Photoreceptor Cell Compartments in the Mouse Retina for Protein Analysis

Rod photoreceptors are highly polarized sensory neurons with distinct compartments. Mouse rods are long (~80 &#181;m) and thin (~2 &#181;m) and are laterally packed in the outermost layer of the retina, the photoreceptor layer, resulting in alignment of analogous subcellular compartments. Traditionally, tangential sectioning of the frozen flat-mounted retina has been used to study the movement and localization of proteins within different rod compartments. However, the high curvature of the rod-dominant mouse retina makes tangential sectioning challenging. Motivated by the study of protein transport between compartments, we developed two peeling methods that reliably isolate the rod outer segment (ROS) and other subcellular compartments for western blots. Our relatively quick and simple techniques deliver enriched and subcellular-specific fractions to quantitatively measure the distribution and redistribution of important photoreceptor proteins in normal rods. Moreover, these isolation techniques can also be easily adapted to isolate and quantitatively investigate the protein composition of other cellular layers within both healthy and degenerating retinae.

This protocol presents two techniques to isolate the subcellular compartments of murine rod photoreceptors for protein analysis. The first method utilizes live retinae and cellulose filter paper to separate rod outer segments, while the second employs lyophilized retinae and adhesive tape to peel away rod inner and outer segment layers.

两种剥离方法，用于分离小鼠视网膜中的光感受器细胞区室以进行蛋白质分析

Rod photoreceptors are highly polarized sensory neurons with distinct compartments. Mouse rods are long (~80 &#181;m) and thin (~2 &#181;m) and are ...