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Cryo-electron Microscopy

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...

Coagulation

Coagulation

The coagulation phase is a critical part of the body's process to prevent blood loss following injury to blood vessels. It involves chemical reactions that form a clot to seal the injured area. The clotting process begins shortly after injury, within 15-20 seconds for severe damage and 1-2 minutes for minor injuries.
During the coagulation phase, clotting factors, or procoagulants, play a vital role in initiating and progressing the coagulation cascade. This cascade is a series of reactions...

Extrinsic and Intrinsic Pathways of Hemostasis

Extrinsic and Intrinsic Pathways of Hemostasis

Blood clotting or coagulation involves extrinsic and intrinsic pathways, which ultimately merge into the common pathway, forming a fibrin clot.
The Extrinsic Pathway
The extrinsic pathway of coagulation is typically initiated by tissue damage that exposes blood to tissue factor (TF), a protein released by the damaged tissue cells outside the blood vessels—this interaction with TF triggers biochemical reactions involving specific clotting factors. The key player here is Factor VII, which...

Protein Folding

Protein Folding

Clot Retraction and Fibrinolysis

Clot Retraction and Fibrinolysis

After a fibrin clot is formed, the next step is clot retraction, a vital process facilitated by platelet contractile proteins, such as actin and myosin. These proteins pull the fibrin strands closer together and condense the clot. This action reduces the size of the clot, creating a smaller, denser structure that effectively seals off the damaged vessel. Clot retraction consolidates the clot and helps with wound healing by bringing the edges of the damaged blood vessel closer together.

Anticoagulant Drugs: Low-Molecular-Weight Heparins

Anticoagulant Drugs: Low-Molecular-Weight Heparins

Hemostasis is a crucial process that prevents excessive blood loss from damaged blood vessels. It involves various mechanisms such as vasoconstriction, platelet adhesion and activation, and fibrin formation. The importance of each mechanism depends on the type of vessel injury. In contrast, thrombosis is the abnormal formation of a blood clot within the blood vessels, leading to potential complications if the clot obstructs blood flow. Thrombosis can be caused by increased coagulability of the...

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Related Experiment Video

Updated: Aug 22, 2025

Helical Organization of Blood Coagulation Factor VIII on Lipid Nanotubes

Helical Organization of Blood Coagulation Factor VIII on Lipid Nanotubes

Published on: June 3, 2014

Cryo-EM structures of coagulation factors.

Enrico Di Cera¹, Bassem M Mohammed¹, Leslie A Pelc¹

¹Edward A. Doisy Department of Biochemistry and Molecular Biology Saint Louis University School of Medicine St. Louis Missouri USA.

Research and Practice in Thrombosis and Haemostasis

|November 9, 2022

Summary

Cryo-electron microscopy (cryo-EM) reveals detailed structures of key blood clotting proteins like factor V and prothrombinase. These cryo-EM structures advance understanding of thrombosis and hemostasis mechanisms.

Keywords:

blood coagulation factor V factor Va prothrombin vitamin K‐dependent clotting factors

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The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells

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Last Updated: Aug 22, 2025

Helical Organization of Blood Coagulation Factor VIII on Lipid Nanotubes

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Published on: June 3, 2014

Do's and Don'ts of Cryo-electron Microscopy: A Primer on Sample Preparation and High Quality Data Collection for Macromolecular 3D Reconstruction

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The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells

The CryoAPEX Method for Electron Microscopy Analysis of Membrane Protein Localization Within Ultrastructurally-Preserved Cells

Published on: February 27, 2020

Area of Science:

Biochemistry
Structural Biology
Hematology

Background:

Coagulation factors are crucial for hemostasis but challenging to study structurally.
Nuclear magnetic resonance (NMR) and X-ray crystallography have limitations for large protein complexes.

Purpose of the Study:

To present the application of cryo-electron microscopy (cryo-EM) for determining structures of coagulation factors.
To highlight advancements in understanding thrombosis and hemostasis through novel structural insights.

Main Methods:

Cryogenic electron microscopy (cryo-EM) was employed to solve protein structures.
Structures of coagulation factors V, Va, prothrombinase, and the prothrombin-prothrombinase complex were determined.

Main Results:

Novel cryo-EM structures of coagulation factors V and Va were obtained.
The structure of prothrombinase, both on nanodiscs and in complex with prothrombin, was solved.
These structures provide molecular insights into prothrombin activation and factor V function.

Conclusions:

Cryo-EM is a powerful technique for investigating large, complex biomolecules like coagulation factors.
The solved structures significantly enhance the understanding of thrombosis and hemostasis.
These findings open new avenues for research in blood coagulation.