Extended Data Fig. 2: Quantification strategy employed to quantify nuclear, perinuclear and cytoplasmic signal using Imaris. | Nature Microbiology

Extended Data Fig. 2: Quantification strategy employed to quantify nuclear, perinuclear and cytoplasmic signal using Imaris.

From: Nuclear pore blockade reveals that HIV-1 completes reverse transcription and uncoating in the nucleus

Extended Data Fig. 2

Schematic representation of the quantification technique used in this manuscript with an example image of a cell expressing the Nup62DG and infected with HIV-1. Cells were stained for the capsid protein p24 and DAPI. a, Nuclear and perinuclear signal was quantified using a DAPI mask generated using the surface function in Imaris that exceeded the boundary of DAPI stain to include the perinuclear signal. As depicted, all signal within this mask was considered nuclear and perinuclear signal. Similarly, all signal outside of this mask was considered cytoplasmic signal. b, To focus on exclusively nuclear signal, an algorithm which reliably identified the nuclear boundary, as indicated by a DAPI stain, was generated in Imaris. Sections close to the upper and lower boundary of the nucleus (in Z) were removed to focus analysis on nuclear events. As in (a) signal within this mask was considered nuclear and all signal outside this mask was considered cytoplasmic and perinuclear.

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