JoVE Logo
Authors
Contact Us

Sign In

A subscription to JoVE is required to view this content. Sign in or start your free trial.

In This Article

  • Overview
  • Protocol
  • Disclosures
  • Materials
  • References

Overview

The video demonstrates a procedure for quantifying the Zika virus in multiple mouse organs via a focus-forming assay. The process involves infecting mammalian cell monolayers with virus-infected tissue homogenates, followed by the use of methyl cellulose to prevent viral spread. Subsequently, immunostaining is employed to enable precise quantification and measurement of organ-specific infectivity.

Protocol

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.

1. Organ Isolation

NOTE: The virus is not stable at room temperature (RT) so the number of animals harvested at one time must be planned carefully to preserve viral titers.

  1. Infect mice using the chosen dose and route, based on the phenotype required. For this protocol, infect 8-10 week-old male and female type I interferon receptor-deficient (Ifnar1-/-) C57BL/6 mice.
    1. Anesthetize Ifnar1....

Access restricted. Please log in or start a trial to view this content.

Disclosures

No conflicts of interest declared.

Materials

NameCompanyCatalog NumberComments
1-bromo-3-chloropropane (BCP)MRC geneBP151
10cc syringeThermo Fisher ScientificBD 309642
18G needleThermo Fisher Scientific22-557-145
1cc TB syringeThermo Fisher Scientific14-823-16H
20cc syringeThermo Fisher Scientific05-561-66
24 tube beadmillThermo Fisher Scientific15 340 163
3.2 mm stainless steel beadsThermo Fisher ScientificNC9084634
37C Tissue Culture incubatorNuair5800
4G2 antibodyin house
96 well flat bottom platesMidsciTP92696
96well round bottom platesMidsciTP92697
Basix 1.5ml eppendorf tubesThermo Fisher Scientific02-682-002
Concentrated Germicidal BleachStaples30966CT
CTL S6 AnalyzerCTLCTL S6 Universal Analyzer
Curved cutting scissorsFine Science Tools14061-11
Dulbecco's Modified Eagle's Medium - high glucose With 4500 mg/L glucoseMilliporeSigmaD5671
Ethanol (molecular biology-grade)MilliporeSigmae7023
Fetal Bovine SerumMilliporeSigmaF0926-500ML
ForcepsFine Science Tools11036-20
Glacial acetic acidMilliporeSigma537020
Goat anti-mouse HRP-labeled antibodyMilliporeSigma8924
HEPES 1 MMilliporeSigmaH3537-100ML
Isopropanol (molecular biology-grade)MilliporeSigmaI9516
Ketamine/Xylazine cocktailComparative Medicine
L-glutamineMilliporeSigmag7513
Magmax RNA purification kitThermo Fisher ScientificAM1830
MethylcelluloseMilliporeSigmaM0512
MicrocentrifugeEpendorf5424R
MiniCollect 0.5ml EDTA tubesBio-one450480
O-ring tubesThermo Fisher Scientific21-403-195
One step q RT-PCR mixThermo Fisher Scientific4392938
ParaformaldehydeThermo Fisher ScientificEMS- 15713-S
Phosphate Buffered SalineMilliporeSigmad8537-500ml
Proline multichannel pipettesSartorius72230/72240
Proline single channel pipettesSartorius728230
RNAse free waterThermo Fisher Scientific10-977-023
RNAzol BDMRC geneRB192
Rocking PlatformThermo Fisher Scientific11-676-333
RPMI 1640FisherMT10040CV
SaponinMilliporeSigmas7900
Spoon/SpatulaFine Science Tools10090-17
Straight cutting scissorsFine Science Tools14060-11
Triton X-100MilliporeSigmat8787
True Blue SubstrateVWR95059-168
TrypsinMilliporeSigmaT3924-100ML

References

....

Access restricted. Please log in or start a trial to view this content.

This article has been published

Video Coming Soon

Source: Brien, J. D. et al., Isolation and Quantification of Zika Virus from Multiple Organs in a Mouse. J. Vis. Exp. (2019)

JoVE Logo

Privacy

Terms of Use

Policies

Contact Us

Recommend to library

JoVE NEWSLETTERS

Research

Education

Authors

Librarians

ABOUT JoVE

Copyright © 2025 MyJoVE Corporation. All rights reserved