MS2(2) (Elsa Anes) Cellular Assays for phagosome maturation: acidification Phagosome maturation blockade by M. tuberculosis

5 Maio 2020, 17:00 • Elsa Anes

MS2(2) (Elsa Anes) Cellular Assays for phagosome maturation: acidification Phagosome maturation blockade by  M. tuberculosis

Introduction

Phagosome maturation: phagolysosome biogenesis

Cellular assays for phagosome maturation

Assays for acidification

Fluorimetry

Fluorescence Microscopy

Fluorophores: FITC, Lysotrackers & lysosensors

Phagosome maturation blockade by M.tuberculosis

Laboratory

Macrophage cell culture. Cellular passage and density.

Macrophage infection with a MOI 10 of Mycobacterium smegmatis.

Labelling acidotrophic vesicles by Lysotracker red.

PFA 9ixation. Slide mounting with 9luoprep.

Image acquisition by Fluorescence microscopy (confocal) by 9luorometry (TECAN) or by 9low cytometry.