Processing of Lentivirus-Transduced Mouse Intestinal Organoids for Immunohistochemical Analysis

Abstract

Source: Van Lidth de Jeude, J. F., et al. A Protocol for Lentiviral Transduction and Downstream Analysis of Intestinal Organoids. J. Vis. Exp. (2015).

This video demonstrates the processing of lentivirus-transduced mouse intestinal organoids for paraffin embedding, to enable their downstream analysis for assessing gene expression.

Protocollo

1. Processing organoids for paraffin embedding and immunohistochemistry

1. Prior to initiation of organoid embedding, pre-warm an aluminum block with holes fitting 12 mm diameter glass tubes in incubator at 70 °C to keep paraffin liquid.

2. Take a single well with full-grown organoids and remove the medium, leaving the embedded organoids intact.

3. Add 1 ml of 4% paraformaldehyde in phosphate-buffered saline (PBS) straight to well and fix at 4 °C anywhere from 1 hr to overnight.

Materiali

Elenco dei materiali utilizzati in questo articolo

Nome	Azienda	Numero di catalogo	Commenti
Paraformaldehyde	Sigma	252549-1L
Glass vial conical 12 mm x 75 mm 5 ml	VWR	LSUKM12
Eosin Yellowish	VWR	1,159,350,025