JoVE Logo

로그인

JoVE 비디오를 활용하시려면 도서관을 통한 기관 구독이 필요합니다.

TIRF Microscopy-Based Visualization of Phagosome Formation and Closure

-- views • 1:27 min

내레이션 대본

Take a polymer-coated glass bottom dish containing IgG-opsonized red blood cells or RBCs adhered to its surface.

Place the dish on a total internal reflection fluorescence microscope stage, maintaining it at physiological temperature.

Add macrophages containing fluorescently labeled cytoplasmic peptides that bind to polymerized actin, facilitating cytoskeleton visualization.

During imaging, direct a laser beam at an angle superior to the critical angle, causing internal reflection and evanescent wave generation at the contact point.

The evanescent waves selectively illuminate the fluorophores at the glass-sample interface, allowing real-time visualization of phagocytosis.

Macrophage Fc receptors bind to the IgG-opsonized RBC, causing receptor clustering around the contact area.

Clustering triggers intracellular signaling pathways and GTPase activation, driving actin polymerization and dynamin recruitment, forming pseudopods.

The pseudopods extend around the RBC, forming a phagocytic cup.

Eventually, the pseudopod tips fuse. The accumulated dynamin mediates phagosome separation from the cell membrane, internalizing the opsonized RBC.

article

02:47

TIRF Microscopy-Based Visualization of Phagosome Formation and Closure

관련 동영상

34 Views

JoVE Logo

개인 정보 보호

이용 약관

정책

연구

교육

JoVE 소개

Copyright © 2025 MyJoVE Corporation. 판권 소유