Retinal Cell Dissociation
Retinal Cell Dissociation - Related Videos
10:20
Single-cell Profiling of Developing and Mature Retinal Neurons
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Cited by 13 •
2012
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.
07:22
Isolation of Retinal Stem Cells from the Mouse Eye
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Cited by 12 •
2010
In this video, we will demonstrate how to isolate retinal stem cells from the ciliary epithelium of the mouse eye and grow them in culture to form clonal retinal spheres. The spheres that are isolated possess the cardinal properties of stem cells: self-renewal and multipotentiality.
11:54
Imaging Exocytosis in Retinal Bipolar Cells with TIRF Microscopy
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Cited by 15 •
2009
In this video, we demonstrate how to label and visualize single synaptic vesicle exocytosis and trafficking in goldfish retinal bipolar cells using total internal reflectance fluorescence (TIRF) microscopy.
05:26
Mechanical Dissociation: A Method to Obtain Viable Cells from a Tissue
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2023
This video describes the non-enzymatic dissociation of fresh human tissue to obtain viable cells. The technique is used for qualitative and quantitative analysis of CD45 positive cells (lymphocytes/leukocytes) present in various normal and malignant human tissues.
11:26
Single-cell RNA-Seq of Defined Subsets of Retinal Ganglion Cells
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Cited by 9 •
2017
Here, we present a combinatorial approach for classifying neuronal cell types prior to isolation and for the subsequent characterization of single-cell transcriptomes. This protocol optimizes the preparation of samples for successful RNA Sequencing (RNA-Seq) and describes a methodology designed specifically for the enhanced understanding of cellular diversity.
FAQs
Enzymatic digestion loosens the extracellular connections that hold retinal cells together, reducing the force needed for separation. Gentle mechanical trituration then releases individual cells or small clusters from the treated tissue. Using these steps in sequence helps produce a preparation containing neurons, glial cells, and other retinal cell types while supporting cellular viability for subsequent analysis.
Gentle trituration helps separate cells after enzymatic treatment without applying unnecessary mechanical stress. This matters because the resulting cells must remain viable for downstream studies such as morphology, gene expression, electrophysiology, imaging, pharmacological assays, or culture. The balance between sufficient dissociation and limited physical disruption influences whether the preparation is suitable for these applications.
A dissociated preparation can contain retinal neurons, glial cells, and other retinal cell types. Examining these populations separately or within small clusters allows researchers to investigate cellular morphology, gene expression, development, and responses to injury or treatment. This is valuable in neuroscience because retinal tissue contains multiple interacting cell types that contribute to retinal circuitry.
A typical workflow begins by treating retinal tissue with enzymes to loosen extracellular connections. The softened tissue is then subjected to gentle mechanical trituration to release individual cells or small clusters. The resulting preparation can be directed toward cell culture, imaging, electrophysiology, gene-expression analysis, or pharmacological testing, depending on the experimental question and the desired cellular outcome.
These preparations provide access to cellular features that can be examined under controlled conditions. Researchers can assess morphology, gene expression, electrophysiological properties, development, and responses to injury or treatment. Because the tissue is separated into cells or small clusters, investigators can connect observed outcomes more directly with particular retinal cell populations or experimental conditions.
The technique is useful when experiments require retinal cells for controlled culture, imaging, pharmacological assays, or regenerative research. It supports investigations of neurodegenerative disease by enabling analysis of cellular responses to injury or treatment. Dissociated material can also contribute to studies of retinal circuitry and to efforts focused on understanding or developing regenerative approaches.