Quantification of fluorescence intensity of antisense constructs within Bodo saltans and its intracellular symbiont

Held, Marie, Ahrar, Mastaneh, Hurst, Gregory and Chrostek, Ewa (2024) Quantification of fluorescence intensity of antisense constructs within Bodo saltans and its intracellular symbiont .

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Description

The purpose of this Protocol is to quantify the intensity of fluorescence resulting from antisense molecules within a microeukaryote Bodo saltans and its intracellular symbiont Candidatus Bodocaedibacter vickermanii1 following incubation. This protocol is specific to two-channel fluorescent images of objects contained within larger objects. It can be used directly to analyse images of objects of similar size. The images were recorded using a confocal laser scanning microscope as single focal plane images, containing three channels: 1) fluorescent antisense molecule, 2) DAPI, 3) transmitted light. The antisense molecule intensity in channel 1 is to be measured. The DAPI channel allows the segmentation of both, the B. saltans nucleus and the symbionts. The transmitted light channel is used for general reference. This Protocol.io contains two sample files (.czi), which open in Fiji and can be re-analyzed using the steps described here. A separate word file with all the steps listed can also be found attached to this Protocol.io.

Item Type:	Other
Uncontrolled Keywords:	Protocol, Workflow, Image analysis
Depositing User:	Symplectic Admin
Date Deposited:	30 Jul 2024 07:16
Last Modified:	30 Jul 2024 07:16
Open Access URL:	https://www.protocols.io/view/quantification-of-fl...
URI:	https://livrepository.liverpool.ac.uk/id/eprint/3183219
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