Imaging and Quantifying Bacterial Infection

Larvae were embedded in 1.5% agarose (low melting point) (Davis and Ramakrishnan, 2009 (link)). A series of z stack images with a 2 μm step size was generated through the infected HBV, using the galvo scanner (laser scanner) of the Nikon A1 confocal microscope with a 20x Plan Apo 0.75 NA objective. Bacterial burdens were determined by using the 3D surface-rendering feature of Imaris (Bitplane Scientific Software) (Yang et al., 2012 (link)).

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Cambier C.J., O’Leary S.M., O’Sullivan M.P., Keane J, & Ramakrishnan L. (2017). Phenolic Glycolipid Facilitates Mycobacterial Escape from Microbicidal Tissue-Resident Macrophages. Immunity, 47(3), 552-565.e4.

Publication 2017

A1 confocal microscope 20x Plan Apo 0.75 NA objective

Agarose Bacterial Confocal microscope Larvae

Corresponding Organization : Trinity College Dublin

Other organizations : Stanford University

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Variable analysis

independent variables

Embedding larvae in 1.5% agarose (low melting point)

dependent variables

Bacterial burdens determined by using the 3D surface-rendering feature of Imaris

control variables

Laser scanner (galvo scanner) of the Nikon A1 confocal microscope with a 20x Plan Apo 0.75 NA objective
Step size of 2 μm for the z-stack images

controls

No positive or negative controls were explicitly mentioned in the provided information.

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