in situET

Jie E Yang1,2,3, Veronika Vrbovská4, Joshua M Mitchell5,6,7

  • 1Department of Biochemistry, University of Wisconsin, Madison, WI, USA.

Nature protocols
|January 30, 2026
PubMed
概括

本研究介绍了使用光显微镜引导的聚焦离子束 (FIB) 研磨的冷电子断层扫描 (cryo-ET) 的简化工作流. 这种方法提高了吞吐量,并保留了用于实地结构生物学的结构数据.

相关概念视频

Cryo-electron Microscopy01:28

Cryo-electron Microscopy

Conventional electron microscopy (EM) involves dehydration, fixation, and staining of biological samples, which distorts the native state of biological molecules and results in several artifacts. Also, the high-energy electron beam damages the sample and makes it difficult to obtain high-resolution images. These issues can be addressed using cryo-EM, which uses frozen samples and gentler electron beams. The technique was developed by Jacques Dubochet, Joachim Frank, and Richard Henderson, for...
4.3K
Focusing of Light in the Eye01:16

Focusing of Light in the Eye

Light rays enter the eye through the cornea, a transparent dome-shaped tissue that is the eye's outermost layer. The cornea bends or refracts, light rays traveling to the pupil. The shape of the cornea determines how much of the light is bent and whether the image will be focused correctly on the retina at the back of the eye. Once the light has passed through both refraction layers, it converges into a single focal point onto a small area. This is where photoreceptors start transforming...
5.9K
Ion Channels01:19

Ion Channels

The movement of ions like sodium, potassium, and calcium into and out of the cell is essential to maintain the electrochemical gradient in living cells. The ion channels—a class of membrane transport proteins—help maintain this ionic gradient for the smooth functioning of physiological activities such as maintaining cell size and volume, conducting nerve impulses, and gas and nutrient exchange.
Ion channels are specialized integral membrane proteins on the plasma membrane that allow...
91.4K
FISH - Fluorescent In-situ Hybridization02:07

FISH - Fluorescent In-situ Hybridization

Fluorescence in situ hybridization, or FISH, was developed in the early 1980s and has quickly become one of the most widely used techniques in cytogenetics. Labeled probes are used to bind complementary DNA or RNA sequences on a chromosome or in a region within a cell. Earlier, the probes could only be obtained by cloning or reverse transcription of a DNA template. Currently, the probe oligonucleotides can be synthesized synthetically. Additionally, with the advancement of optical techniques,...
24.4K
Confocal Fluorescence Microscopy01:16

Confocal Fluorescence Microscopy

Confocal microscopy is an advanced microscopic technique. The prime advantage of the confocal microscope over other microscopy techniques is its ability to block the out-of-focus light from the illuminated samples using pinholes. It is widely used with fluorescence optics to obtain high-resolution, sharp contrast images. Unlike optical microscopes, confocal microscopes use a focused beam of light laser to scan the entire sample surface at different z-planes. These microscopes are, therefore,...
21.1K
Beams01:30

Beams

Beams are integral components of structural engineering and construction, designed to support loads applied at various points along their length. These long, straight members can be classified based on geometry, cross-section, support type, and equilibrium condition.
Based on geometry, beams can be straight, tapered, or curved. Straight beams are the most common type and have a constant cross-section throughout their length. Tapered beams, on the other hand, have a varying cross-section along...
1.9K