NDLI: A simple method for multiday imaging of slice cultures.

Content Provider	World Health Organization (WHO)-Global Index Medicus
Author	Seidl, Armin H. Rubel, Edwin W.
Description	Country affiliation: United States Author Affiliation: Seidl AH ( Department of Otolaryngology-HNS, Virginia Merrill Bloedel Hearing Research Center, University of Washington, Seattle, Washington 98195-7923, USA.)
Abstract	The organotypic slice culture (Stoppini et al. A simple method for organotypic cultures of nervous tissue. 1991;37:173-182) has become the method of choice to answer a variety of questions in neuroscience. For many experiments, however, it would be beneficial to image or manipulate a slice culture repeatedly, for example, over the course of many days. We prepared organotypic slice cultures of the auditory brainstem of P3 and P4 mice and kept them in vitro for up to 4 weeks. Single cells in the auditory brainstem were transfected with plasmids expressing fluorescent proteins by way of electroporation (Haas et al. Single-cell electroporation for gene transfer in vivo. 2001;29:583-591). The culture was then placed in a chamber perfused with oxygenated ACSF and the labeled cell imaged with an inverted wide-field microscope repeatedly for multiple days, recording several time-points per day, before returning the slice to the incubator. We describe a simple method to image a slice culture preparation during the course of multiple days and over many continuous hours, without noticeable damage to the tissue or photobleaching. Our method uses a simple, inexpensive custom-built insulator constructed around the microscope to maintain controlled temperature and uses a perfusion chamber as used for in vitro slice recordings.
File Format	HTM / HTML
ISSN	1059910X
e-ISSN	10970029
DOI	10.1002/jemt.20750
Journal	Microscopy Research and Technique
Issue Number	1
Volume Number	73
Language	English
Publisher	Wiley-Liss
Publisher Date	2010-01-01
Publisher Place	United States
Access Restriction	Open
Subject Keyword	Discipline Chemistry Techniques Brain Stem Cytology Image Processing, Computer-assisted Microscopy, Video Animals Genes, Reporter Luminescent Proteins Genetics Metabolism Mice Mice, Inbred C57bl Organ Culture Techniques Plasmids Staining And Labeling Transfection Research Support, N.i.h., Extramural Research Support, Non-u.s. Gov't
Content Type	Text
Resource Type	Article
Subject	Anatomy Histology Instrumentation Medical Laboratory Technology

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1	Ministry of Education (GoI), Department of Higher Education	Sanctioning Authority	https://www.education.gov.in/ict-initiatives
2	Indian Institute of Technology Kharagpur	Host Institute of the Project: The host institute of the project is responsible for providing infrastructure support and hosting the project	https://www.iitkgp.ac.in
3	National Digital Library of India Office, Indian Institute of Technology Kharagpur	The administrative and infrastructural headquarters of the project	Dr. B. Sutradhar bsutra@ndl.gov.in
4	Project PI / Joint PI	Principal Investigator and Joint Principal Investigators of the project	Dr. B. Sutradhar bsutra@ndl.gov.in Prof. Saswat Chakrabarti will be added soon
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