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Imaging of Human Immunodeficiency Virus Proteolytic Maturation by stimulated emission depletion microscopy (CAT#: STEM-MIT-0353-LJX)

Introduction

Concomitant with human immunodeficiency virus type 1 (HIV-1) budding from a host cell, cleavage of the structural Gag polyproteins by the viral protease (PR) triggers complete remodeling of virion architecture. This maturation process is essential for virus infectivity. Electron tomography provided structures of immature and mature HIV-1 with a diameter of 120-140 nm, but information about the sequence and dynamics of structural rearrangements is lacking. The service employed super-resolution STED (stimulated emission depletion) fluorescence nanoscopy of HIV-1 carrying labeled Gag to visualize the virion architecture. The incomplete Gag lattice of immature virions was clearly distinguishable from the condensed distribution of mature protein subunits.




Principle

Stimulated emission depletion (STED) microscopy uses two light sources. One source emits light that excites the fluorophores, and the other emits a ring laser of different wavelengths, which is used to suppress fluorescence.

Applications

Imaging of the intensity distribution of the fluorescent sample
Imaging of living samples
Measuring of the fluorescence lifetime and fluorescence correlation spectrum of the fluorescent samples
Used in the fields of biology, medicine and materials science

Procedure

1. Sampling
2. Preparation of slices
3. Staining (Select according to the specific experimental situation)
4. Observation

Materials

• Sample Type:
Human Immunodeficiency Virus Protein

Notes

Operate in strict accordance with the operating procedures, and shall not arbitrarily change the operating procedures
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