Related Experiment Video

Updated: Feb 3, 2026

Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells
09:47

Generation of Retinal Organoids from Healthy and Retinal Disease-Specific Human-Induced Pluripotent Stem Cells

Published on: December 9, 2022

4.5K

Immuno-TEM/STEM in Retinal Research

Sanae Sakami1, Krzysztof Palczewski2

  • 1Department of Pharmacology, Cleveland Center for Membrane and Structural Biology, Case Western Reserve University School of Medicine, Cleveland, OH, USA.

Methods in Molecular Biology (Clifton, N.J.)
|October 17, 2018
PubMed

Insights

High-angle annular dark field scanning transmission electron microscopy (HAADF-STEM) enhances contrast in immuno-electron microscopy (immuno-EM) specimens. This method clearly visualizes photoreceptor disk membranes, improving ultrastructural protein localization studies.

Area of Science:

  • Electron Microscopy
  • Cell Biology
  • Biochemistry

Background:

  • Immunoelectron microscopy (Immuno-EM) is crucial for ultrastructural protein localization.
  • Standard immuno-EM often suffers from low contrast in membrane structures due to fixation without osmium.
  • Observing membrane details at the ultrastructural level remains a challenge.

Purpose of the Study:

  • To improve the visibility of membrane structures in immuno-EM specimens.
  • To demonstrate the utility of HAADF-STEM for immuno-EM analysis.
  • To establish a method for enhanced visualization of protein localization at membrane interfaces.

Main Methods:

  • Utilized high-angle annular dark field (HAADF)-scanning transmission electron microscopy (STEM).
  • Applied HAADF-STEM for the observation of immuno-EM samples.
  • Compared HAADF-STEM imaging with conventional transmission electron microscopy (TEM) for immuno-EM.

Main Results:

  • Photoreceptor disk membranes were clearly visible using HAADF-STEM.
  • Significantly enhanced contrast of membrane structures was achieved.
  • The method proved effective for visualizing otherwise low-contrast membrane features.

Conclusions:

  • HAADF-STEM offers superior contrast for membrane structures in immuno-EM.
  • This technique can be extended to visualize various other membrane structures.
  • Enhanced visualization aids in accurate ultrastructural protein localization studies.

Keywords:
Disk (disc) membraneHAADF-STEMImmuno-EMLR white resin and rhodopsinPhotoreceptorTEM

More Related Videos

Isolation of Retinal Stem Cells from the Mouse Eye07:22

Isolation of Retinal Stem Cells from the Mouse Eye

Published on: September 11, 2010

25.4K
Efficient Derivation of Retinal Pigment Epithelium Cells from Stem Cells07:07

Efficient Derivation of Retinal Pigment Epithelium Cells from Stem Cells

Published on: March 8, 2015

9.7K

Related Concept Videos

Induced Pluripotent Stem Cells01:13

Induced Pluripotent Stem Cells

Stem cells are undifferentiated cells that divide and produce different types of cells. Ordinarily, cells that have differentiated into a specific cell type are post-mitotic—that is, they no longer divide. However, scientists have found a way to reprogram these mature cells so that they “de-differentiate” and return to an unspecialized, proliferative state. These cells are also pluripotent like embryonic stem cells—able to produce all cell types—and are therefore...
28.0K
Induced Pluripotent Stem Cells01:06

Induced Pluripotent Stem Cells

Stem cells are undifferentiated cells that divide and produce different cell types. Ordinarily, cells that have differentiated into a specific cell type are terminally differentiated; however, scientists have found a way to reprogram these mature cells so that they dedifferentiate and return to an unspecialized, proliferative state. These cells are pluripotent like embryonic stem cells—able to produce all cell types—and are called induced pluripotent stem cells (iPSCs).
Somatic...
5.6K
Adult Stem Cells01:33

Adult Stem Cells

Stem cells are undifferentiated cells that divide and produce more stem cells or progenitor cells that differentiate into mature, specialized cell types. All the cells in the body are generated from stem cells in the early embryo, but small populations of stem cells are also present in many adult tissues including the bone marrow, brain, skin, and gut. These adult stem cells typically produce the various cell types found in that tissue—to replace cells that are damaged or to continuously...
33.9K
Embryonic Stem Cells00:58

Embryonic Stem Cells

Embryonic stem (ES) cells are undifferentiated pluripotent cells, meaning they can produce any cell type in the body. This gives them tremendous potential in science and medicine since they can generate specific cell types for use in research or to replace body cells lost due to damage or disease.
32.4K
Embryonic Stem Cells00:57

Embryonic Stem Cells

Embryonic stem (ES) cells were first discovered in mice in 1981 by Martin Evans. In 1998, James Thomson identified a method to isolate embryonic stem cells from humans. Human embryonic stem cells (hESCs) are obtained from 3-5 day old embryos that remain unused after an in vitro fertilization procedure.
ES cells are grown in a culture medium where they can divide indefinitely, creating ES cell lines. Under certain conditions, ES cells can differentiate, either spontaneously into a variety of...
5.0K
Distinctive Features of Adult Stem Cells vs Cancer Stem Cells01:18

Distinctive Features of Adult Stem Cells vs Cancer Stem Cells

A stem cell is an unspecialized cell that can divide without limit as needed and can, under specific conditions, differentiate into specialized cells.
Adult stem cells
Adult stem cells are tissue-specific; hence, they divide to develop the tissue from which they originate. One type of adult stem cell is the epithelial stem cell, which gives rise to the keratinocytes in the multiple layers of epithelial cells in the epidermis of the skin. Adult bone marrow has three distinct types of stem cells:...
4.5K